-^^2.^0 N87 Columbia anitJem'tp^^P/^ mtljfCttpoflmgork College of ^fjpsiicianfl! anb S'urgeonjS ILibrarp GiJ^-fc of Dr. C. F. M&.cPoTva-!-.. . -A8c^,?) Entered according to Act of Congress, in the year 1912, by LEA & FEBIGER in the Office of the Librarian of Congress. All rights reserved. ^ 2_ TLo ms ffatbet 2)r« I£^num& IRoijee Mboee long aiiD active professional career bas been Devoted, witbout personal ambition or selfisb advancement, to tbe 0OOD ot tbe Dental profession, anO wbose unselfisbness anD sacrifice bave maDe possible all tbat "IT bave C>one or mas accomplisb PREFACE It is indispensable for the successful treatment of disease in the dental tissues that the dentist should acquire as intimate a knowledge of structure as is essential to the physician, and consequently that a parallel study of his- tology should be followed. The development of biology has placed histolog}' at the basis of all the medical sciences, for as, in the last analysis, all physiology is cell physiology, and all pathology is cell pathology, a knowledge of structure and function is necessary for an intelligent conception of the workings of the animal body in health and for the restoration of normal function when impaired by disease. Yet as recently as fifteen years ago, when the author began teaching Dental Histology in the Northwestern University, the subject was comparatively new in the cur- riculum, and was considered rather unimportant and as having little practical value. It would be impossible to give adequate acknowledgment to the help received from Dr. G. V. Black in developing the course. Every detail was worked out in the closest cooperation with him, and for years he guided and directed the work. The object of a course in general and special histology suited to the needs of dental students is to convey a definite knowledge of the activities of these parts of the human bod}' in terms of tissues and cells. This is the basis of every prac- tical procedure. The structure of the enamel and dentine is obviously the starting point in handling these tissues and in the preparation of cavities, and the structure and function of the pulp, the bone, the periosteum, and the peridental membrane are similarly the basis for an understanding of vi PREFACE their pathology and treatment. The study of the enamel in relation to cavity preparation has proved to be of the greatest value not only in forming better cavity walls, but also in facilitating operation. For this purpose it is necessary to understand both the structure of the enamel in itself, and the arrangement of the structural elements in relation to the tooth crown. To accomplish this, sections cut through the crown in various planes must be studied and their relation to it kept in mind. The modern dentist while looking at the surface of a tooth must think of the enamel in terms of its structural elements, and use this knowledge in handling the tissue. In the following pages the enamel is studied primarily in relation to operative dentistry. The chapters on the pulp, peridental membrane, and periosteum are likewise intended to emphasize the relation of structure to function, and to impress the idea that the treatment of disease in these tissues is in every instance a biological problem. In forming true conceptions of caries and necrosis, a knowledge of the intercellular substances and their relation to the cells in the structure and function of tissues is necessary. A chapter has, therefore, been devoted to this subject. The study of the structure and development of bone has very greatly modified treatment in orthodontia, as it is now recognized that in all movements of the teeth the results are accomplished by tissue changes under the influence of mechanical stimuli. Though there are many good books on general histology, they are not fully adapted to the special needs of the dental curriculum. The author has accordingly felt that teachers and students of dentistry might find some advantage in a work covering the subject from their own standpoint, and embodying the results of his experience in teaching as well as in research. In a word, this volume has been planned primarily as a text-book for use in dental schools, and it aims to provide students with a didactic text and teachers with a course to follow. It contains directions for twenty- two days of laboratory work, and an appendix giving tech- nical methods for preparation of material for the classes. PREFACE vil It suggests many fields for ot-iginal investigation, and tech- nical directions which would enable any man to begin such work. Let us hope that the benefits certain to result from discoveries still to be made will lead some students and practitioners to interest themselves in this inviting field. Most of the illustrations are from the author's own nega- tives. Those on the relation of enamel structure to cavity walls are new as well as original in plan. The drawings illustrating the periosteum and pathological conditions of the pulp were made by Dr. G. V. Black, and he has writ- ten the chapter on his machine for making ground sections and the technique of its use. Some illustrations are taken from other works, and these are duly credited in every instance. Thanks are also due to Dr. Louis Schmidt, of the Rockefeller Institute, who made the colored plates from the author's specimens, and to A. B. Streetdain, of the University of Chicago, for his work on some difficult diagrams. Finally, the author wishes to thank his publishers for their pains and patience in carrying out his wishes. F. B. N. Chicago, 1912. CONTENTS INTRODUCTION 17 CHAPTER I Homologies 19 CHAPTER II The Dextal Tissues 28 CHAPTER HI The Examel 38 CHAPTER IV The Structural Elemexts of the Examel 43 CHAPTER V Characteristics of the Examel Tissue 52 CHAPTER VI The Directiox of the Examel Rods ix^ the Tooth Crowx . 65 CHAPTER VII The Relatiox of the Structure to the Cuttixg of the Examel , 73 CHAPTER VIII The Structural Requiremexts for Stroxg ExNTamel Walls . 80 CHAPTER IX The Preparatiox of Typical Examel Walls 89 CHAPTER X Structural Defects ix the Enamel .,.,.... 107 X CONTENTS CHAPTER XI Special Areas of Weakness for Enamel Margins . . . 124 CHAPTER XII The Effect of Caries on the Structure of the Enamel . . 143 CHAPTER XIII The Dentine 167 CHAPTER XIV The Cementum 188 CHAPTER XV Dental Pulp 201 CHAPTER XVI Structural Changes in the Pathology of the Pulp . . . 219 CHAPTER XVII Intercellular Substances 236 CHAPTER XVIII Bone 247 CHAPTER XIX Bone Formation and Growth 255 CHAPTER XX Periosteum 262 CHAPTER XXI The Attachment of the Teeth 271 CHAPTER XXII Peridental Membrane 279 CHAPTER XXIII The Cellular Elements of the Peridental Membrane. . 294 CHAPTER XXIV The Mouth Cavity 323 CONTENTS XI CHAPTER XXV Biological Considerations Fundamental to Embryology . 335 CHAPTER XXVI Early Stages of Embryology 340 CHAPTER XXVII The Development of the Tooth Germ 362 CHAPTER XXVIII Th:e Relation of the Teeth to the Development of the Face 374 PART II DIRECTIONS FOR LABORATORY WORK (Twenty-four Periods in the Laboratory) Period I 429 Period II 429 Period III 432 Period IV 435 Period V 435 Period VI 437 Period VII 439 Period VIII 440 Period IX 440 Period X 441 Period XI 442 Period XII 442 Period XIII 443 Period XIV 444 Period XV 445 Period XVI 445 Period XVII 446 Period XVIII 446 Period XIX 447 Period XX 448 Period XXI 449 Period XXII 450 Period XXIII 451 Period XXIV > 442 xii CONTENTS APPENDIX CHAPTER I The Grinding of Microscopic Specimens, using the Grind- ing Machine 453 CHAPTER II The Theory of Histological Technique 478 CHAPTER III General Histological Methods 485 CHAPTER IV Fixing Agents and Staining Solutions 496 DENTAL HISTOLOGY INTRODUCTION The development in knowledge of the cell has had a most profound effect upon the entire practice of medicine; in fact, the progress of modern medicine has dated from the studies of cell biology, the germ theory of disease being only one of the phases of this development. In terms of the cell theory the functions of the body are but the manifest expression of the -activities of thousands or millions of more or less independent but correlated centres of activity. If these centres or cells perform their functions correctly, the functions of the body are normal, but if they fail to perform their office or work abnormally, the functions of the body are perverted. In the last analysis, then, all physiology is cell physiology, all pathology cell pathology. To modern medicine, histology, or the cell structure of the organs and tissues of the body, together with cell physiology, is the rational foundation of all practice. This is as true for the dentist as for the physician in regard to the soft tissues of the mouth and teeth that he is called upon to handle. With caries of the teeth, the disease which most demands the attention of the dentist, the case is somewhat different. Caries of the teeth is an active destruction, by outside agencies, of a formed material which is the result of cell activity, the teeth themselves being passive. The cellular activities of organs and tissues of the body may have an influence, but this is only in producing those conditions of environment which render the activities of the destructive 18 DENTAL HISTOLOGY agent efficient in their action upon the tooth tissues. Though the dental tissues are passive, the phenomena of caries can only be understood when the structure of the tissues is understood, and not only must the treatment be based upon knowledge of the structure of the tissues, but the mechanical execution of the treatment is facilitated by that knowledge of structure. In the preparation of cavities, the arrangement of the enamel wall is determined by the knowledge of the direction of the enamel prisms in that locality, and to a certain extent the position of cavity margins must be governed by the knowledge of the structure of the enamel. In the execution of the work a minute knowledge of the direction of enamel rods becomes the most important element in rapidity and success of operation. The longer the author studies and teaches the structure of the enamel in its relation to the structure and preparation of enamel walls, the more he finds himself using this knowledge at the chair in daily operations. He believes that nothing will do more to increase facility, rapidity, and success of operation than a close study of the enamel structure. All tissues are made up of two structural elements — cells and intercellular substances. The cells give the vital char- acteristics, the intercellular substances the physical character. The cells are the active living elements, the intercellular substances are formed materials produced by the activity of the cells, and more or less dependent upon them to main- tain their quality, but they possess no vital properties. They surround and support the cells, and the physical character- istics are given by them. An understanding of the relation of cells and intercellular substances in the structure and function of tissues is absolutely fundamental to a study of dental histology, and should be acquired in a thorough study of general histology before the subject is undertaken. CHAPTER I HOMOLOGIES Exoskeleton. — In studying the organization of animal forms they are found, very early in the evolutionary stages, to de- velop some sort of a framework, or skeleton, to support and protect the creature. In the lower and earlier forms this framework is formed entirely of some sort of shell upon the outside of the creature, and consequently is called an exo- skeleton. This may be either horny or chitinous in nature, as in the insects, crabs, etc., or it may be calcified, as in the shell-fish, or it may be both. The exoskeleton serves not only as a supporting framework, but also as a protection. Endoskeleton. — In the higher forms an internal framework, or endoskeleton, is developed, which forms the scaffolding to support the creature, but does not act as a protection. In the first place, this is of cartilage, but may be changed into bone. In lower forms of animals it remains always cartilage. In man the cartilage is partly converted into bone, all of the bones of the endoskeleton being preceded by cartilage. The first trace of the endoskeleton is found in the lowest form of vertebrate, the Amphioxus or Lancit, the lowest form of fish, and appears as a rod or notochord in the dorsal region. There is also an important difference in the nervous organization (Figs. 1 and 2). In the invertebrate the nervous system is represented by a larger or smaller ganglion in the anterior or head end, corresponding to the brain; this is dorsal to the alimentary canal. From this a ring passes around the anterior end of the alimentary canal and unites with a chain of ganglia ventral to it. The nervous system of the invertebrate then is, with the exception of the brain ganglia, ventral to the alimentary canal, and corresponds to a - ^ I -JB -S s g § allflli S s I § i - S ^- - ' ft " „ § 2 s t a g- ^: 2 so . •■ « o "a. 'r o 2 ® ^ "3 .2 S _ -sj _p Ji ft ^ « fl "^ -S -^ « ^: !: s ■J "-'!-2l I'M ^ci'Scift.'"-^ . ^ ^ - . s «. g §• :d^^£ 22 HOMOLOGIES the sympathetic system in higher animals. It will be noted that this arrangement puts the nervous system, which controls the activity of the individual, in the most protected position. The invertebrate crawling upon the ground is subject to attack or injury from above, but it may be cut almost in two before the nervous system is reached. In the vertebrate the central nervous system appears as a chain of ganglia dorsal to the alimentary canal and notochord (Fig. 2). This difference is significant, and may be expressed roughly in this way : The invertebrate framework is an out- side protecting shell, upon which the creature depends for protection. The vertebrate framework is an internal struc- ture to facilitate motion and give support, and is accompanied by a development of the nervous organization, so that the creature protects itself by more rapid motion. In the inverte- brate the digestive system is above or dorsal to the nervous system, in the vertebrate the nervous system is in the upper position, both structurally and functionally. In ascending in the scale of organization the endoskeleton increases in importance and development, while the exo- skeleton decreases in importance and development. From the standpoint of comparative anatomy the teeth are not a part of the osseous system, but appendages of the skin, and are to be compared with such structures in the body as the hair and the nails. The teeth aie a part of the exo- skeleton, and their relation to the bones of the endoskeleton is entirely secondary for the purpose of strength, the bone growing up around the tooth to support it. When the skin of such an animal as the shark is examined, the entire sur- face is found covered with small calcified bodies, which are really small, simple, cone-shaped teeth. From the stand- point of development the mouth cavity is to be regarded as a part of the outside surface of the body which has been enclosed by the development of neighboring parts, and the dermal scales, or rudimentary teeth, which are found in the skin covering the arches forming the jaws, have undergone special development for the purpose of seizing and masti- cating the animal's food. In the simplest forms there is only HOMOLOGY AND ANALOGY 23 a development in size and shape of these scales, and they are supported only by the connective tissue which underlies the skin. These teeth are easily torn off in the attempt to hold a resisting prey, and in the shark (Fig. 3) they are continually being replaced by new ones. In the more highly developed forms, the bone forming the jaw grows upward around the bases of these scale-like teeth to support them more firmlv and render them more useful. Fig. 3 Shark's skull (Lamna coinubica), showing succession of teeth. Homology and Analogy. — In biology structures that are similar in formation and origin are called homologous. Structures that are similar in function are called analogous. A structure or organ may be both homologous and analo- gous to another, but not necessarily so. For instance, the wing of a fly is analogous to the wing of a bird, because they are used for the same purpose, but they are not homologous. The wing of a bat and the wing of a bird are both analogous and homologous, being used for the same purpose, and 24 HOMOLOGIES having similar structure and origin. The arm of man is homologous to the wing of a bird but not analogous to it. The jaws of a crab or beetle are analogous to the jaws of man, but they are not homologous structures, as the jaws of the crabs and insects are modified legs. The teeth are said to Fig. 4 ,-^5^3^???; Development of the hair: Sc, stratum corneum; SM. stratum malpighii; C, derma; F, follicle; Dr, sebaceous gland; CZ, central, PZ, peripheral zone of hair germ; HK, hair knob; P, beginning the formation ol the hair papilla; P', same in a later stage of development when it has become vascular. (Wiedersheim, Comparative Anatomy of Vertebrates.) be homologous to the dermal scales of certain fishes, and to the appendages of the skin, such as the hair and nails, because they are similar in structure and origin (Plate I). Comparison of Structure. — If the tooth is compared with the hair in this way this will be better understood. The hair PLATE I Comparison of Structure of Tooth and Hair. COMPARISON OF ORIGIN 25 may be considered as a horny structure composed of epithelial cells resting upon a papilla of connective tissue. The tooth may be considered a calcified structure, formed by epithelial cells, resting upon a papilla of connective tissue, which is also partially calcified. Comparison of Origin. — From a study of the development of the tooth and the hair, the similarity of their origin and structure be- comes more apparent. The first step in the develop- ment of the hair is a thickening of the epithelium at a point, the epithelial cells multiplying and growing down into the connective tissue below, so as to make a two-layered bag or cap, the con- nective tissue groAving up in the form of a cone-shaped papilla into the cavity of the cap (Fig. 4). The epithelial cells of the inner layer, next to the connec- tive tissue, multiply rapidly and develop horny material and are pushed out from the surface of the skin as the shaft of the hair. Diugram to illustrate development of a tooth: A. inner layer of enamel germ; B, outer layer; C, remains of intermediate cells; D, dentine; Z)./, dental lamina; ^.epithelium; E.G, enamel germ; En, enamel; F, dental furrow; L.D, labiodental furrow; M, con- nective-tissue cells; O, odontoblasts: P, den- tine papillaf R.G, reserve germ; V, blood- vessel. (Cunningham's Anatomy.) Fig. 5 III f'> V \ ■ 26 HOMOLOGIES In the development of the tooth there is at first a thicken- ing of the epithehum, and a mass of epithehal cells like that forming the hair, hut larger, grows down into the connective tissue (Fig. 5). This becomes bulbous, then invaginated, forming a two-layered cap. The two layers are at first per- fect and are farther from the surface than the epithelial Fig. 6 Changes in the mandible with buccal and lingual view. structure which develops the hair. A cone-shaped papilla of connective tissue, the dental papilla, grows up into the cavity of the epithehal organ corresponding to the bulb of the hair. The inner layer of epithelial cells produce the enamel, the outer layer of connective-tissue cells, covering the connective- RELATION TO THE BONE 27 tissue papilla, develop the dentine, leaving the pulp inside as the remains of the dental papilla. Relation to the Bone. — The relation of the bones of the jaws to the teeth is entirely secondary and transient. They grow up around the roots of the teeth to support them, and are destroyed and removed with the loss of the teeth or the cessation of their function. In this way the development of the alveolar process appears around the roots of the tem- porary teeth. All this bone surrounding their roots is ab- sorbed and removed with the loss of the temporary denti- tion, and a new alveolar process grows up around the roots of the permanent teeth as they are formed. This develop- ment of bone around the roots of the teeth leads to the changes in the shape of the body of the lower jaw, increasing the thickness from the mental foramen and the inferior dental canal upward (Fig. 6) . When the teeth are finally lost this bone is again removed and the body of the jaw is reduced in thickness from above downward. These phenomena have an important bearing upon the causes and treatment of diseased conditions of the teeth, particularly those which involve the supporting tissues. CHAPTER II THE DENTAL TISSUES Study of the structure of the teeth shows that all teeth, from the simplest to the most complex, are composed of but four tissues — enamel, dentine, cementum, and the pulp, or formative tissue of the dentine. Even the simplest placoid scales, as found in the skin of the shark and dog-fish, contain these four tissues. In many of the specialized forms of teeth some of these tissues may be absent. For instance, in the bony fishes the teeth are fastened to the bone by an interlocking of bone and dentine, forming an ankylosed attachment, and the cementum is absent; but in some of these there is also a slight formation of cementum. In the tusks of elephants during the func- tional period the dentine is not covered by enamel, but when the tusk first erupted there was a slight enamel cap, which was at once broken or worn off. In many instances the enamel seems to be entirely absent, and for that reason it has sometimes been called the most inconstant of the dental tissues, but in every case in which the development of the tooth has been studied an enamel organ has been found. It is probably much more nearly correct to consider that in all cases enamel is formed, but that it may be so thin and transparent as to be very difficult to recognize, and very soon may be entirely lost. • FUNCTIONS OF THE DENTAL TISSUES The Enamel. — The enamel forms a hard protecting surface or cap especially adapted to resist abrasion. It is the hardest animal tissue, but brittle and inelastic, and dependent upon FUNCTIONS OF THE DENTAL TISSUES 29 the support of the elastic dentine for strength. Its func- tion is to resist the abrasion of friction. Its arrangement in many instances is found specially modified for this purpose. The Dentine. — The dentine is the strong elastic tissue form- ing the great mass of the tooth, and gives to it its strength. Teeth that are subjected to stress and force are often made up of dentine without enamel. If, for instance, the tusks of the elephant used for such purposes as tearing down branches, spading up the ground, and so on, were made up entirely of enamel, they would break off the first time they were locked in the branches or driven into the ground, but the elastic dentine gives and bends and will stand great stress. The teeth of many animals which use their tusks in fighting are constructed on the same plan. The Cementum. — The cementum furnishes attachment for the connective-tissue fibers which fasten the tooth to the bone or surrounding tissues. It is formed on the enamel and dentine both before and after the eruption of the teeth. The formation of the cementum on the surface of the root fastens the surrounding connective-tissue fibers to the tooth. The fibers are calcified along with the matrix of the cementum which is built up around them. These fibres in man and the higher animals extend to the bone and the surrounding tissues and support the teeth against the forces of occlusion, and hold the surrounding tissues in proper relation to the teeth. The function of the cementum is, therefore, to attach the connective tissue fibers to the surface of the root. The Pulp. — The pulp is the remains of the formative organ of the dentine. In teeth of continuous growth it remains actively functional throughout the life of the tooth, but in teeth of limited grow^th, after the typical development of dentine, it becomes functional again only in response to irritations, which, however, may be local or reflex. The pulp performs two functions — a vital function, the formation of dentine, and a sensory function, the response to thermal change. Summaxy. — The dental tissues, i. e., enamel, dentine, cementum, and pulp, are so called not simply because they 30 THE DENTAL TISSUES are found in the human teeth, but because all teeth are composed of these four tissues. It is true that in comparative dental histology consider- able difference exists in the microscopic structure of these tissues from the teeth of different animals, but certain characteristics are very persistent and quite characteristic of each. DISTRIBUTION OF THE DENTAL TISSUES The arrangement and distribution of the dental tissues in the structure of the human teeth is best studied in ground sections cut longitudinally through the entire tooth (Plate II), and series of transverse sections cut through the roots. For this purpose the sections should not be too thin (from 10 to 20 microns). For the study of the arrangement of the cementum and dentine in the roots at least three transverse sections should be ground from each root, one from the gingival, one from the middle, and one from the apical third. The Enamel. — The enamel forms a cap over the exposed por- tion of the tooth. Its function is to resist the abrasions of mas- tication. It gives the detail of crown form to the tooth. It extends to the gingival line, and, except in old age, is covered in the gingival portions by the epithelium of the gingivus, which lies in contact with it but is not attached to it. It is thin in the gingival portion and is normally overlapped slightly by the cementum at the gingival line. It extends farther apically on the labial and lingual, and buccal and lingual, than upon the proximal surfaces, especially on the incisors, cuspids, and bicuspids. It is thickest in the occlusal third of the axial surfaces, and on the occlusal surfaces of the molars and bicuspids, especially over the cusps. In the incisors and cuspids it is thickest in the occlusal third on the labial and over the marginal ridges on the lingual and the dento-enamel junction, which, though not parallel with the surface of the enamel, is usually curved in the same direction. In the molars and bicuspids the dento-enamel junction in < 5. W I ? O ft - r T (t o 0. ft D z: c r > PI DISTRIBUTION OF THE DENTAL TISSUES 31 the occlusal thirds on the buccal and lingual is usually curved in the opposite direction. That is, while the surface Fin. : { v.- \ I, ,-i: i . i I.: C»:^^ >4i4 I 32 THE DENTAL TISSUES of the enamel is convex, the surface of the dentine is concave. It will be seen that this not only gives a greater thickness to the enamel in the region which will resist abrasion, but also gives it a firmer seat upon the dentine. (Study illustrations in Chapter X.) The dento-enamel junction is seldom a smooth, even surface, but will appear scalloped in sections, projections of dentine extending between projections of enamel (Fig. 7). In three dimensions this means that rounded projections of the enamel rest in rounded depressions of the dentine surface, and pointed projections of the dentine extend between the rounded pro- jections of the enamel. This is similar but much less marked than the interlocking of the papilla of connective tissue with the projections of the Malpighian layer of stratified squamous epithelium of the skin and mucous membrane. In some cases these projections of dentine into the enamel may be quite marked. This scalloping of the dento-enamel junc- tion gives a stronger attachment of the enamel to the den- tine, and accounts, partially at least, for the difference that is observed in the ease with which enamel can be removed from the dentine in the preparation of roots for crowns. Where the two tissues join with smooth surfaces the enamel can be comparatively easily cleaved away; Avhere the scalloping is marked it is removed with much greater difficulty. The Dentine. — The dentine gives the strength to the tooth. This should never be lost sight of in operations, and sound dentine should always be conserved to the greatest possible extent in the preparation of cavities. That the function of the dentine is to give strength will be seen more clearly from a comparative study of teeth modified for special functions. The dentine forms the greatest mass of the tooth, the type form being determined by it. The cusps and ridges, although different in form, are still represented in the dentine as well as the number and shape of the roots, while the detail of the form of the roots is modified by the addition of the cementum on the surface. The dentine forms a layer of comparatively even thickness DISTRIBUTION OF THE DENTAL TISSUES 33 surrounding the central cavity or pulp chamber, which is occupied by the formative organ. From this cavity a great number of small tubules extend through the calcified dentine matrix to the surface under the enamel and cementum. In the crown portion the course of these tubules is characteris- tically curved like the letter S or /, so that the tubules tend to enter the pulp chamber at right angles to the surface and to end under the enamel at right angles to the dento-enamel junction (Plate II). On closer study these tubule directions will be found to be more complicated, but in studying the distribution of dentine they should be noted. In the root portion the tubules are usually comparatively straight, that is, without the double curve, and are at about right angles to the axis of the canal. The outer layer of dentine under low magnification pre- sents a peculiar granular appearance, which is specially apparent under the cementum. This is known as the granular layer of Tomes, and is caused by irregular spaces in the dentine matrix which communicate with the dentinal tubules. The Cementum. — The cementum covers the dentine in the root portion, and in most cases slightly overlaps the enamel at the gingival line. This is not always true, for in some cases it just meets the enamel, and in others there is a space where the dentine is uncovered between the enamel and the cementum (Fig. 8). It has not been positively determined whether this can ever be considered a normal condition, and the author has some reason to suppose that the sec- tions showing this condition were from teeth from which the gums had receded and the cementum was destroyed. The sensitiveness which is so marked in some cases, where the gums have receded beyond the gingival line, is probably due to the loss of cementum and the uncovering of the granular layer of Tomes. The cementum is thin and structureless in appearance in the gingival portion when viewed with low powers, but becomes thicker in the apical third. In the thicker portions irregular spaces (lacunae) with radiating canals (canaliculi) 3 34 THE DENTAL TISSUES are seen. In life these spaces contain living cells (the cement corpuscles), which correspond to the bone corpuscles found in the lacuna of bone. Upon the convex surfaces of the root the cementum is thin; upon the concave surface it is thicker. This increases with age, and so the continuous formation of cementum tends to round the outlines of the roots and to Fig. 8 Gingival line, showing the relation of enamel and cementum. unite them where they approach each other. The fibers which are built in the cementum are often imperfectly calcified, especially where the layers are thick, so that in the ground sections they may often be easily mistaken for canals, because the imperfectly calcified fiber has shrunken in the preparation. ADAPT A TION IN DISTRIBUTION OF DENTAL TISSUES 35 ADAPTATION IN THE DISTRIBUTION OF DENTAL TISSUES If the teeth of mammals are studied in a comparative way many modifications will be found in the relative amount and distribution of the dental tissues, adapting the tooth to per- form special functions. A study of these modified or special- ized teeth will give a better understanding of the functions of the tissues in the tooth. The human tooth may be taken as a type of omnivorous tooth, and the arrangement and distribution of its tissues has already been described. Teeth of Continuous Growth. — In many animals the teeth or some special teeth are developed as weapons for use in fighting, or as implements to aid in securing food. It is usually the cuspid teeth that show this modification, as in the tusks of the boar and many species of the carnivora, the tusks of the walrus, and other examples. In the case of the elephant the incisors have been developed in the same way. Whenever the teeth have been developed in size for uses which require strength and the ability to withstand stress and strain, the increase in size is by development of the mass of dentine, the enamel often being entirely lost during the functional period. If these teeth were composed chiefly of enamel they would be too brittle. These tusks, which, as in the case of the elephant, sometimes reach a weight of many hundreds of pounds, are usually deeply embedded in the bone, and the concealed portion is covered with a layer of cementum which attaches the fibers, holding them to the bone, but they retain a conical pulp in a cone-shaped pulp chamber at the base of the tooth, which continues to form dentine. The tooth is pushed out of the socket, as the shaft of the hair is pushed out, by the multiplication of cells covering the bulb. In this way the size of the tooth is maintained as the exposed and functional part is worn off. Strength and elasticity are required, therefore the dentine is developed. The cementum which is formed on the 36 THE DENTAL TISSUES embedded portion for attachment of fibers is worn off as soon as it is exposed to friction. Chisel Teeth. — The incisors of the rodents, as rats, mice, squirrels, and beavers, present an interesting modification for a special function. These teeth are used as chisels for cutting hard substances, as wood, shells of nuts, etc. Here strength and hardness are required. The dentine is increased by the continual function of a conical persistent pulp which continues to form dentine, and the enamel organ is carried down into the socket, to the base of the dental papilla, on the labial, instead of stopping at the gingival line, as in the human incisors. In this position it continues to build enamel on the labial side of the dentine. The enamel rods, instead of being straight, are twisted about each other in a complicated fashion, giving the maximum of hardness. As the incisors work against each other by the movements of the jaw, the dentine is worn off on the lingual side and the enamel kept in the form of a chisel edge. There is also a modification of the temporomandibular articulation, allowing the lower jaw to move forward and back as well as up and down, but not laterally, so that the lower incisors can be closed either lingually or labrally to the upper, and in this way both the upper and the lower incisors are made to sharpen each other in use. In this case there is need for both strength and hardness, and both dentine and enamel are continuously being formed at the base of the tooth embedded in the socket, and the cementum is formed over the embedded portions as the medium of attachment. Grinding Teeth. — In a grinding tooth, as in the molar of the horse and cow, and in a much more complicated form in the elephant, the three tissues — enamel, cementum, and dentine — are arranged so as to form, by the different rapidity of abrasion, corrugated grinding surfaces like millstones. The conditions can be understood if it is remembered that the cusps in the dentine are very high, and are covered by a comparatively thin layer of enamel. After the enamel is formed, and while the tooth is embedded in its crypt in the bone, cementum is formed, covering the surface and filling ADAPTA TION IN DISTRIBUTION OF DENTAL TISSUES 37 up the hollows between the cusps, so that the crown when it first erupts is rounded, with only enamel showing at the tips of the cusps. As soon as the tooth wears, the tip of the enamel is worn through, so that the circumference of the crown shows first cementum, then enamel, then dentine, then enamel, then cementum, then enamel, and so on. The foldings of the enamel often become very complicated, but the most complicated forms can be understood in this way. In describing the structure of the teeth and the arrange- ment of the structural elements of the tissues, directions are described with reference to three planes: The mesio-disto- axial plane passing through the centre of the crown from mesial to distal and parallel with the long axis of the tooth. The bucco-linguo-axial plane, a plane passing through the centre of the crown from buccal to lingual and parallel with the long axis of the tooth. The horizontal plane at right angles to the axial planes. CHAPTER III THE ENAMEL The enamel differs from all other calcified tissues: 1. In origin. 2. In degree of calcification. 3. In relation to its formative organ. 4. In the form of the structural elements of the tissue. It is well to emphasize these points of difference, for throughout dental and medical writing, reasoning by analogy from bone conditions to tooth conditions, and especially to changes in the enamel, is often found. For instance, the argument has been made that because there may be changes in the bones in pregnancy, "softening" of the teeth would be expected. Many similar though less crude arguments would not be made if it were remembered that histologically, histogenetically, physiologically, and morphologically the enamel stands alone. Origin. — The enamel is the onl}^ calcified tissue derived from the epithelium. All other calcified tissues are con- nective tissues. Histogenetically, then, the enamel is ultimately derived from the epiblastic germ layer, while all other calcified tissues arise from the mesoblast. Thus, even at the first step in the differentiation of cells, the enamel is different and independent from bone, cementum, or dentine. It is natural, therefore, to find the enamel differing from bone in every other respect. On the other hand, the relation of the enamel to the epithelium becomes more and more apparent. For instance, imperfections in the structure of the enamel during its formation are most likely to be pro- duced by systemic conditions which affect the epithelium. The eruptive fevers occurring during enamel formation often • DEGREE OF CALCIFICATION 39 produce imperfections of structure. Scarlet fever is most pronounced in its epithelial effect, causing loss of skin, loss of living epithelium of the alimentary tract, and often loss of hair, and is likewise most likely to produce pitted and atrophied teeth. In other words, the same poison which is produced by the germ of scarlet fever causes the death of epithelial cells, of the skin, of the hair bulb, of the mucous membrane, and of the enamel organ. The most recent work of Dr. Black shows the brown and mottled enamel of certain localities to be found associated with greatly freckled skin. Enamel, therefore, must be considered as epithelial in origin and ultimately from the epiblast, while all other calcified tissues are connective tissue and ultimately of mesoblastic origin. Degree of Calcification. — The enamel is by far the hardest animal tissue. Chemically it is composed of water, calcium phosphate, carbonate, and a small amount of fluoride, magnesium phosphate, and a trace of other salts. Normally it should contain no organic matter. Von Bibra gives the following analysis : Calcium phosphate and fluoride 89 . 82 Calcium carbonate 4.37 ^Magnesium phosphate 1.34 Other salts 0.88 Cartilage 3.39 Fat 0.20 It is very difficult to obtain enamel for chemical analysis entirely free from dentine, and small portions of dentine clinging to it are probably responsible for some of the organic matter given in the above analysis. In all the older analyses the enamel was said to contain 95 to 97 per cent, of inorganic matter, and 3 to 5 per cent, of organic matter, while the percentage in dentine was given as 72 per cent, of inorganic and 28 per cent, of organic, and in bone as 68 per cent, inorganic and 32 per cent, organic (dry compact bone). This in itself shows an enormous difference in the degree of calcification between enamel and the other hard tissues, but the results of more recent work are 40 THE ENAMEL still more remarkable. In most of the original studies of the chemical composition, the enamel was broken into small pieces and dried for some time at a temperature above the boiling point of water, to drive off all the moisture. The dry enamel was weighed and then ignited, and the loss in weight taken as the amount of organic matter. In 1896 Dr. Charles Tomes,^ of London, published the results of his chemical analysis of enamel, in which he showed that a large part of the loss of weight in ignition was due to the loss of water. He carried out ignition in tubes to collect the products of combustion, and found that between red and white heat from 2 to 3 per cent, of water was given off. This occurred suddenly and with almost explosive violence, blowing large pieces to fragments. While this did not account entirely for all of the matter previously considered organic, the character of the product of combustion and the observation of the material during ignition led him to conclude that the remaining portion was due to the dentine adhering to the enamel, and that the enamel contained not more than a trace of organic matter. Dr. Leon Williams attacked the problem from the micro- scopic and microchemical side, and was forced to the con- clusion that normal enamel contains no organic matter. No trace of organic matter can be found in sections of enamel by staining. And if the enamel is dissolved by acid and the progress observed, not a trace of organic matrix can be found. The conclusion is therefore imperative that enamel is composed entirely of inorganic matter, which has been deposited and calcified in the form of the tissue by the formative cells. In other words, enamel is formed material produced by cells and laid down in a definite structure, but it contains no organic matrix, while all other calcified tissues are composed of an organic matrix of ultimate fibrous and gelatin-yielding character, in which inorganic salts are deposited in a weak chemical combination, and living cells are retained in spaces of the formed material. 1 Journal of Physiology. RELATION TO THE FORMATIVE TISSUE 41 If bone or dentine is subjected to the action of acid, the combination between the organic and inorganic matter is broken up and the inorganic matter dissolved, leaving the organic portion, which yields gelatin when boiled in water, in the form of the original tissue. If enamel is treated with acid the cementing substance between the rods is first attacked and is dissolved more rapidly, then the rods are attacked from their sides, and finally the tissue is entirely destroyed, leaving no trace of structure. Apparently the greater the dilution of the acid the greater will be the extent of the solution of the cementing substance before the rods are destroyed. If bone or dentine is burned or ignited, the organic matter will be driven off and the inorganic portion will be left in the form of the tissue, still showing its structure. If enamel is ignited, water of combination and whatever foreign matter has clung to the pieces is given off, but the form of the tissue is unchanged. To illustrate the difl^er- ence by a crude comparison: Bone matrix may be likened to a piece of cloth into which organic salts have been deposited until it has become stifle and rigid, but the web of the cloth is still seen. The salts may be dissolved out and the cloth left, or the cloth may be burned out and the salts left. The enamel may be compared to a fossil in which, by molecular change, the organic matter has been removed and inorganic matter substituted, so that no organic matter remains, but the structure is preserved. If the inorganic salts were dissolved, no trace of structure would remain. On the other hand, by ignition, nothing but water can be driven oft'. Relation to the Formative Tissue. — The enamel is produced by epithelial cells, which are lost and destroyed after the tissue is completed. Any such thing, therefore, as a vital change in the tissue is biologically unthinkable. After the enamel is formed it can be changed only by chemical and physical action of its environment. All other calcified tissues are formed by connective tissue, and remain in vital relation with connective tissue of undifter- 42 THE ENAMEL entiated character. Bone and dentine matrix are, therefore, simply calcified intercellular substances containing living cells in the spaces of the matrix, which maintain its chemical quality. A change in the character or amount of the matrix might possibl}^, therefore, be brought about by the vital activit}^ of these cells. Moreover, the formed matrix is always in vital relation with undifferentiated connective tissue, which may at any time destroy or rebuild it. There is, therefore, no basis for comparison between pathologic conditions of bone and enamel. The Form of the Structural Elements. — The enamel is made up of prismatic rods of inorganic matter, held together by an inorganic cementing substance. All other calcified tissues are made up of fibrous intercellular substance, con- taining inorganic salts and usually arranged in layers. The structure of the enamel differs so greatly from all other calcified tissues that it is difficult to compare them briefly. CHAPTER IV THE STRUCTURAL ELEMENTS OF THE ENAMEL The enamel is composed of two structural elements: 1. The enamel rods or prisms, sometimes called enamel fibers. 2. The interprismatic, or cementing substance. Fig. 9 £na JO X) Enamel Rods.— The enamel rods are long slender prismatic rods irregularly five or sLx sided and alternately expanded and constricted throughout their length (Figs. 9 and 10). They are from three and four-tenths to four and five-tenths microns 44 THE STRUCTURAL ELEMENTS OF THE ENAMEL in diameter, and many of them extend from the dento-enamel junction to the surface of the enamel. They are of the same diameter at their outer and inner ends. This last statement is emphasized, as the direct opposite is stated in some stand- ard text-books of histology. In the formation of the tissue they are arranged so that the expansions in adjoining rods come opposite to each other, and do not interlock with the constrictions, so that there is alternately a greater and a less amount of cementing substance between them. Fig. 10 Enamel rods isolated by scraping. (About SOO X) It is evident that the outer surface of the enamel is much greater than the surface of the dentine at the dento-enamel junction. This greater area is obtained in two ways: 1. The rods are at right angles to the dentine at the dento-enamel junction, but are seldom at right angles to the outer surface. This may be illustrated by bending the leaves ENAMEL RODS 45 of a book, or cutting a stack of paper obliquely. The sheets of paper are of the same thickness, but when cut at right angles to the sheets the area of the cut surface is not so great as when the leaves are cut diagonally. 2. Many of the enamel rods undoubtedly extend from the dento-enamel junction to the surface of the enamel, though it is difficult to follow individual rods through this distance, Fig. 11 Enamel rods in thin etched section. (About 800 X) but there are also short rods which extend from the surface part way to the dentine. These short rods end in tapering points between converging rods that extend the entire dis- tance. The short rods are specially numerous in the most convex portion of the surface, as over the tips of the cusps, occlusal edges, and marginal ridges. These areas, therefore, become of special importance in connection with the forma- 46 THE STRUCTURAL ELEMENTS OF THE ENAMEL tion of enamel walls, as will be considered in detail later on (Fig. 11). Differences between Enamel Rods and Cementing Substance. ■ — AYliile the cementing substance and the substance of the rods are both entirely inorganic, or, more correctly, are com- posed entirely of mineral salts, they differ in physical and chemical properties as follows : 1. The cementing substance is not as strong as the pris- matic substance. 2. The cementing substance is more readily soluble in dilute acids than the rod substance. 3. The cementing substance is of slightly different (greater) refracting index than the substance of the rod. The author wishes to emphasize these statements, as the exact opposite is found in some of the standard texts, at least concerning the first and second statements. The facts are, however, so easily demonstrable that anyone may satisfy himself without difficulty. Relative Strength of the Enamel Rods and the Cementing Substance. — The cementing substance is not as strong as the substance of the rods. The most striking characteristics of the enamel, and the first to attract the attention of the student and the operator, are its hardness and its tendency to split or cleave in certain directions. On examination it is found that this is determined by the direction of the rods, and is caused by the difference in strength between the two substances. Sections ground at right angles to the rod direction are very difficult to prepare because of the tendency of the section to break to pieces. If a section that is beginning to crack (Fig. 12) is studied, the crack is found to follow the line of the cementing sub- stance running around the rods. In some places a rod may be split through its centre, but most of the rods remain perfect, and the cementing substance breaks. In the same way a section cut in the direction of the rods shows the crack following the lines of the cementing substance (Fig. 13), here and there breaking across a few rods, and then fol- lowing the direction again; but the rods separate on the RELATIVE STRENGTH OF THE ENAMEL RODS 47 line of union, not at the centres of the rods. This fact becomes fundamental in the cutting of enamel and in the preparation of strong enamel walls. Fig. 12 Transverse section of enamel rods. (.About bO X) Fig. 13 Enamel showing direction of cleavage. (About 70 X) 48 THE STRUCTURAL ELEMENTS OF THE ENAMEL Relative Solubility of Enamel Rods and Cementing Sub- stance. — If a thin section of enamel cut parallel witli the direction of the enamel rods is mounted in water and hydro- chloric acid (2 per cent.) is allowed to run under the cover- glass and the action observed, it will be seen to attack the cementing substance more rapidly, dissolving it out from between the enamel rods and attacking their sides. If the action is stopped the ends of the rods will be seen pro- jecting like the pickets of a fence, as shown in the photograph (Fig. 14). The more dilute the acid the greater will be the distance to which the cementing substance is removed before the rods are destroyed. Fig. 14 The effect of acid on a section of enamel. Etching. — If a section of enamel is ground at right angles to the direction of the rods, mounted in glycerin and photo- graphed, the outline of the rods will be seen with difficulty (Fig. 15). The refracting index of the two substances is so nearly the same that the section seems of almost uniform transparency. The thinner the section, the greater will be the difficulty of recognizing the rods. Oblique illumina- tion and the use of a small diaphragm will, however, resolve them. If the section is washed and treated with 2 per cent, hydrochloric acid for a few seconds, washed, and remounted in glycerin, the rods are distinctly outlined (Fig. 16). The acid attacks the cementing substance and the surface of the section is etched as if an engraving tool had been run RELATIVE SOLUBILITY OF THE EX AM EL RODS 49 around the rods. The fine grooves on the surface refract the fight and outfine the rods. The difference in appear- ance in longitudinal sections, that is, sections parallel with the direction of enamel rods, is quite as striking. For the study of enamel rod directions this etching is of the greatest impor- tance. Only one side of the section should be acted upon Enamel ground at right angles to the rods. Not treated with acid. (About 500 X) by the acid, and the section should be mounted etched side up. If etched upon both surfaces, the grooves in the lower surface cannot be in focus at the same time as those of the upper surface and will blur the definition. The difference in the solubility of the rods and cementing substance is beautifully illustrated in the effect of caries 4 50 THE STRUCTURAL ELEMENTS OF THE ENAMEL on the structure of the enamel (see ilhistrations in Chai)ter XII), and caries of the enamel cannot be understood unless these fundamental facts are remembered. The question, "AVhat causes the difference in solubility between the enamel rods and the cementing substance?" cannot be satisfactorily answered at the i)rcsent time. While both the rods and the Fig. 10 The same section as Fig. 15 after treatment with acid. (About 500 X) cementing substance are normally composed entirely of inorganic salts, there may be different salts in the two sub- stances, or the salts may be in different physical condition. There is great need for careful work in this field. Recent work has strongly emphasized the distinctness of the two structural elements of the enamel. DIFFERENCE IN REFRACTING INDEX 51 First, the study of the beginnings of caries of the enamel, and the effect of caries upon the structure of the enamel, brought out the difference in solubility in acids and showed the extent of tissue injury before a cavity is formed. Later, the study of atrophy developed the fact that certain patho- logic or abnormal conditions may hinder or entirely pre- vent the formation of the rods while the cementing sub- stance is formed, and still more recently the investigation of dystrophies of the enamel occurring in certain prescribed localities, showed perfect rod formation and entire absence of the cementing substance. These facts suggest the hypoth- esis that the enamel rods and the cementing substance have a different origin, or are formed by different cells, and that pathological conditions may prevent the formation of one and not the other. In view of these factors it is very neces- sary that a new investigation of the process of enamel for- mation be undertaken, as present knowledge of the process does not explain such conditions. Difference in Refracting Index between the Rods and the Cementing Substance. — The cementing substance is of slightly greater refracting index than the substance of the rods. If it were not for this it would be impossible to see the rods in unetched sections, either longitudinal or transverse. The appearance of striation seen in longitudinal sections is also dependent upon this difference in action on transmitted light. CHAPTER V CHARACTERISTICS OF THE ENAMEL TISSUE From what has been said of the structural elements of the tissue, their physical and chemical properties, and their arrangement in the tissue, it is apparent that the striking characteristics of the enamel are the result of these factors; and that it can be intelligently dealt with only by thinking of it always in these terms. Enamel showing cleavage. The enamel may be crudely compared to a pavement made up of tall columns closely cemented together by an inorganic cement. The wear comes on the ends of the columns, and they furnish great resistance to the abrasion of friction. STRAIGHT ENAMEL 53 When supported upon a good and elastic foundation it is very difficult to break it down, but when an opening has been made in it, and the foundation removed from underneath, the columns are comparatively easily split off and tumbled into the opening (Fig. 17). This figure is crude, but it is a very helpful one in learning to think of the enamel in terms of its structural elements. Fig. 18 Straight enamel rods. (About 80 X) Straight Enamel. — Upon the axial surfaces of the teeth the rods are usually straight and parallel with each other, and most of them extend from the dentine to the surface. Such enamel will split or cleave in the direction of the rods with 54 CHARACTERISTICS OF THE ENAMEL TISSUE comparative ease, and breaks down very readily when the dentine is removed from under it. It will usually cleave through its entire thickness and break away from sound dentine when properly attacked with sharp hand instru- ments. Such enamel is called straight enamel, as con- trasted with gnarled enamel. It is best illustrated by cutting sections lal)iolingualIy through the incisors, though there is consi(leral)le variation in different teeth (Figs. 13 and 18). Fig. 19 Gnarled enamel. (About 80 >0 Gnarled Enamel. — Upon the occlusal surfaces of the molars and bicuspids, and especially over the tips of cusps and marginal ridges, the rods are seldom straight and GNARLED ENAMEL 55 parallel through the thickness of the enamel, but are wound and twisted about each other, especially in the deeper half toward the dento-enamel junction. This is known as gnarled enamel, and its appearance is in marked contrast with straight enamel. Fig. 20 Gnarled enamel. (About 50 X) Toward the surface the rods are usually straight and parallel for a longer or shorter distance, but as the dento- enamel junction is approached they become twisted. This is true of most of the occlusal surfaces of molars and bicus- pids, but the gnarled condition extends farther toward the surface-over the tips of the cusps, or the point at which the rods were first completed in the growth of the crown. As 5G CHARACTERISTICS OF THE ENAMEL TISSUE cleavage is caused by the difference in the strength of the rods and cementing substance, it is easy to see that gnarled enamel will not split or cleave easily when resting upon sound dentine. This is often encountered in extending occlusal cavities. The straight portion will split, but where the rods begin to twist they break off, leaving a portion resting on the dentine w-hich will resist the attack of any cutting instrument from the surface (Figs. 19, 20, and 21). Fig, 21 Gnarled enamel from etched section. (About 100 X) The Effect of Structure on the Cutting of Enamel. — The two kinds of enamel may be compared to straight-grained pine wood and a pine knot. The first will split easily in the direction of the fibers, the latter will split only in an irregu- lar way and with the greatest difficulty. This difference in the arrangements of the structural elements leads to the difference in the feeling of various teeth to cutting instru- ments, and is the basis for the clinical experience of hard and soft teeth. It is not a matter of degree of calcification, APPEARANCES CHARACTERISTIC OF ENAMEL 57 but the arrangement of the structural elements, and gnarled enamel will break down as rapidly under the effect of caries as straight enamel would. From a study of the positions in which the rods are usually twisted about each other, and those in which they are usually straight, it seems probable that the twisting is due to movements in the dental papilla and the enamel organ during the formation of the tissue. These movements may be produced by variations in the blood pressure which cause oscillations, or shifting of the tissues on each other. These differences in the arrangement of the structural elements of the enamel must be constantly kept in mind, and will be referred to many times in connection with the use of cutting instruments on the enamel and the preparation of cavity walls. APPEARANCES CHARACTERISTIC OF ENAMEL Striation. — Striation is the appearance of fine light and dark markings occurring alternately in the length of the enamel rods. This is not unlike the striation of voluntary muscle fibers, and has a similar cause. It is seen both in thin sections cut in the direction of the rods, and in isolated enamel rods. It is caused by the alternate expansions and constrictions of the rods and the difference in the refracting index between the rods and the cementing substance. If isolated rods (Fig. 22) are observed with a -g^ or yV objec- tive, they will be seen to be marked by alternate light and dark areas across the rods; on changing the focus up and down, the light and dark areas will change places, just as in looking at a red blood corpuscle the centre may appear dark and the rim light, or the centre light and the rim dark, depending upon the exactness of focus. This is caused by the refraction of the light as it passes through the convex and concave portions of the rod. If the cementing substance were of exactly the same refracting index as the rods, when the rods were fastened together in the tissue there would be no appearajice of striation, but as it is not, refraction of light 58 CHARACTERISTICS OF THE ENAMEL TISSUE occurs in ])assiiig from rod substance to cementing sub- stance, and the striation is apparent in sections. There is considerable difference in the distinctness of striation in different sections of enamel. This is probably due to the fact that the cementing substance has more nearly the same refracting index as the rods in some specimens. When the formation of enamel has been studied it will be found that the enamel rods have been formed by globules which are Fig 22 ^ 3^\ Isolated enaniL'i r')d^<. iAiMun 1000 X.) deposited one on top of the other to form the rods, and the cementing substance fills up the space. The globules in the adjacent rods come opposite each other, so that there is alternately a greater and a less amount of cementing substance between the rods. Each cross-mark, therefore, represents a globule deposited in the formation of the rod, and striation may be said to be a record of the growth of the individual rods (Figs. 23 and 24). APPEARANCES CHARACTERISTIC OF ENAMEL 59 Fig. 23 1 1 ■ ■ ■ i B ? ^B ^M H ■ "-■■ JH ^^H w^ ^^ '^H m^Hi ^K^ m - ^ H^l iv . ;.' ' flrv' r m B U ';--> rJH ^Ktt_ -: .v^ E&nT'V i hm ■ H^ «■ ■ ^^B H| 1 — ^^ ^•,-" .' '"- •. •-; / .^ ■ ,-. ,. ^^^i MB ■ IKHI IMH IHIH Knamei showing both striation and stratification. (About SO X) Fig. 24 Enamel showing striation. (About 1000 X) 60 CHARACTERISTICS OF THE ENAMEL TISSUE Imperfections in the cementing substance render the striation more apparent because they increase the difference in refraction between the two substances. The action of acid either upon isolated rods or upon sections renders striation more apparent because it attacks the cementing substance faster than the globules forming the rods, and therefore increases the refraction. Von Beber has claimed that the appearance of striation was caused by the action of acid on the section, and that even in mounting in balsam the acidity of the balsam affected the tissue. It is true that any action of acid increases the distinctness of the cross- striation, but it is not the cause of it. Stratification, or the Bands of Retzius. — If longitudinal sections of moderate thickness are observed with the low power, brownish bands are seen running through the enamel, which suggests the appearance of stratification in rocks. These were first described by Retzius and were named after him — the brown bands or striae of Retzius. A better name would be incremental lines. The bands of Retzius, or incremental lines, are caused by actual coloring matter which is deposited with the inorganic salts in the formation of the tissue. They are, therefore, best seen with low powers and in sections that are not too thin. In sections that are thinner than the diameter of a single rod, or less than four microns, they become almost invisible. For the study of the bands of Retzius sections should be ground labiolingually through the incisors, bucco- lingually through the bicuspids and molars, striking the centre at the cusps. They may be studied also in mesiodistal sections, but the sections should be in such a direction as to be at right angles to the zones. Fig. 25 shows the tip of an incisor in which the bands are very well marked. They are seen to begin at the dento-enamel junction on the incisal edge, and sweep in larger and larger zones around this point. Each band represents what was at one time the sur- face of the enamel already formed, and the line upon which formation was progressing. They are, therefore, truly incremental lines. The zones reach the surface of the APPEARANCES CHARACTERISTIC OF ENAMEL 61 enamel first at the point over the centre of beginning calcifi- cation, and the succeeding bands extend from the surface Fig. 25 Tip of an incisor. (About 50 X) of the enamel, near the occlusal, to the dento-enamel junction much farther apically, and corresponding lines are seen on opposite sides of the section. In Fig. 26 the band which C)2 CHARACTERISTICS OF THE ENAMEL TISSUE is at the surface at A and /I' reaches the dento-enamel junc- tion at B and B\ This means that when the enamel rods which form the surface at A were completed, the rods at Fig. 26 Incisor tip showing stratification or incremental lines. Rods at A were fully formed at the time the rods at B were Vjeginning to form. (About 50 X) B were just beginning to be formed at the dento-enamel junction. A layer of functioning ameloblasts occupied this position. The bands of Retzius are always curved APPEARANCES CHARACTERISTIC OF ENAMEL 63 and usually pass obliquely across the enamel rods, but are parallel neither with the dento-enamel junction or the surface of the enamel. As they pass toward the gingival Fig. 27 Stratification of enamel: the cusp of a bicuspid: De. dento-enamel junction: Ed, enamel defecj showing in the heavj' stratification band; Ig, interglobular spaces in the dentine. (About 40 X) 64 CHARACTERISTICS OF THE ENAMEL TISSUE the angle which they form with the axis of the tooth becomes greater. Any disturbance of nutrition which affects the formation of enamel is always shown in the increased distinct- ness of the bands (Fig. 27). The bands of Retzius, therefore, form a record of the formation of the tissue, and by their study the points of beginning calcification and the manner of the development of the tooth crown may be followed. This will be con- sidered again in connection with the grooves, pits, and natural defects of the enamel. Fig. 28 ^ Lines of Schreger. (About 5 X) Lines of Schreger. — These are lines appearing in the enamel extending from the dento-enamel junction to or toward the surface. They are caused by the direction in which the enamel rods are cut. They may be seen in sections, but are best shown by photographing the cut surface of the enamel by reflected light and with very low magnification. The rods are twisting about each other, and in one streak they are cut longitudinally, in the next obliquely, and the alternations of these directions cause the appearance of the lines (Fig. 28). CHAPTER VI THE DIRECTION OF THE ENAMEL RODS IN THE TOOTH CROWN In describing the direction of the enamel rods and their arrangement in what may be called the architecture of the tooth crown, they are always considered as extending from the dento-enamel junction outward. This is not only con- venient, but logical, as they are formed in that way, beginning at the dento-enamel junction and being completed at the surface. Enamel is formed from within outward, the cells which produce it lying outside of the tissue already formed, and there are many things about the arrangement of the rods and their relation to each other that are understood only when this is borne in mind. The direction of the enamel rods is described by referring them to the horizontal and axial planes, which have been previously defined (page 37). The centigrade scale, that is the division of the circle into one hundred equal arcs, is used because those familiar with instrument nomenclature are already familiar with these angles, and readily picture them.i When a rod is said to be inclined 12 centigrades 1 In the centigrade division the circle is divided into one hundred parts, each called a centigrade. One centigrade is equal to 3.6 degrees of the astronomical circle. 25 centi- grades to 90 degrees, 12f cen- tigrades to 45 degrees. The cut gives a comparison of the two systems of measuring angles 2 70 180'' Centigrade di\'ision. GO DIRECTION OF ENAMEL RODS IN TOOTH CROWN occlusally from tlie horizontal plane, it means that if a plane at right angles to the long axis of the tooth is passed through the end of the rod at the dento-enamel junction, the rod will lie to the occlusal of it and form an angle of 12 cen- tigrades with it. In the same way, if a rod is said to be inclined 12 centigrades buccally from the mesiodistal plane, it means that if a plane parallel with the axis of the tooth, and extending from mesio to distal, is passed through the end of a rod at the dento-enamel junction, the rod will lie to the buccal of it, and form an angle of 12 centigrades with it. By a little practice with these terms the direction of the enamel rods can be very easily and clearly pictured to the mind. The General Direction of Enamel Rods. — The general direc- tion of the enamel rods has been variously described by different authors, but all of these general statements are very imperfect and often misleading. For instance, they are sometimes said to radiate from the centre of the crown or the pulp chamber, but it will be seen that this does not apply to the rods which form the lingual slopes of the buccal cusps, or the buccal slopes of the lingual cusps of bicuspids and molars. Again, they have been said to be, in general, perpendicular to the surface, but it will be found from the study of sections that there are very few places upon the surface where this is true, and that in many places they are far from perpendicu- lar to the surface. From a stud}^ of sections it wdll be seen that the general arrangement of enamel rods, in the archi- tecture of the tooth crown is such as to give the greatest strength to the perfect tissue, and to furnish the greatest resist- ance to abrasion in the use of the teeth for mastication. In a buccolingual section through a bicuspid (Fig. 29), beginning at the gingival line, the enamel is normally slightly overlapped by the cementum, and in the gingival third the rods are inclined more or less apically from the horizontal plane. The degree of inclination varies considerably. It may be as much as 12 centigrades, but is usually not more than 6. In general, the more convex the surface the greater will THE GENERAL DIRECTION OF ENAMEL RODS 67 be the inclination. At some point between the junction of the gingival and middle thirds and the middle of the middle third of the surface they are in the horizontal plane and at right angles to the axis of the tooth, and at this point they are usually very nearly perpendicular to the surface. Passing Fig. 29 Diagram of enamel rod directions, from a photograph of a buccolingual section of an upper bicuspid. occlusally from this point, they incline more and more occlusally until in the occlusal third they reach an inclina- tion of 18 to 20 centigrades occlusalh' from the horizontal. The rods which form the tip of the buccal cusps do not reach the tip of the dentine cusp, but the buccal slope of 68 DIRECTION OF ENAMEL RODS IN TOOTH CROWN the dentine. This becomes important, as will be seen later. Over the tip of the dentine cusp the rods are in the axial plane, but in this position they are usually very much twisted. Passing down the lingual slope, they become more and more inclined lingually from the mesiodistal axial plane. Fig. 30 Diagram of enamel rod directions, drawn from a mesiodistal section of a bicuspid. and the degree of inclination is related to the height of the cusp — the taller the cusp the greater the inclination. At the developmental groove or pit they meet the rods of the lingual cusp, which are inclined in the opposite direction. In a mesiodistal section (Fig. 30) the plan of arrangement will be seen to be the same, the tip of the marginal ridge corresponding to the tip of the cusp. In an incisor the THE GENERAL DIRECTION OF ENAMEL RODS 69 I Fig. 31 1 Disturbance of enamel rod directions on labial surface of a cuspid. (About SO X 70 DIRECTION OF ENAMEL RODS IN TOOTH CROWN Fia. 32 Disturbance of enamel rod directions on lingual surface of same tooth as Fig. 33. (About 80 X) SPECIAL AREAS 71 arrangement is similar, the lingual marginal ridge corre- sponding to a rudimentary cusp. This general plan should be studied in several sections of the various classes of teeth before the rod direction is studied more minutely. Effect of Atrophy. — Whenever an atrophy groove appears upon the surface, the rod directions will be found to be more or less disturbed. Fig. 31 shows a position on the labial surface of a cuspid. In this position the disturbance of the enamel rod direction is very marked. The rods tend to be in whirls and the structure is more or less deficient. On the lingual side of the same section (Fig. 32) the disturbance in structure is so great that it is difficult to make out the rod direction. Many such areas will be found in sections. Some condition which has affected the nutrition of the enamel- forming cells results in a local disturbance of the structural elements. SPECIAL AREAS The Gingival Third. — There is much variation in enamel rod direction in difierent teeth as the gingival line is approached. The inclination apically from the horizontal may be very great, as much as 12 to 15 centigrades in some instances, as in Fig. 33, but this is exceptional. It may be very slight, or the rods may be almost in the horizontal plane. The direction of the rods in these areas become very important in the preparation of the gingival wall of proximal cavities, and cavities in the gingival third of buccal and labial surfaces. The Tips of the Cusps. — In studying the rod directions in the region of the cusps and marginal ridges, it must be borne in mind that the formation of enamel begins at the dento-enamel junction, at separate points, and that the growth is recorded in the tissue by the bands of Retzius, each band having been at one time the surface of the enamel cap then formed. In a buccolingual section the formation of the buccal and lingual cusps will be shown (Chapter X). While the little caps are growing they are being carried apart by the growth of the dental papilla and enamel organ, until the calcifications unite at the dento-enamel 72 DIRECTION OF ENAMEL RODS IN TOOTH CROWN junction. When this occurs the dental papilla has reached its maximum mesio-distal diameter. The enamel organ, however, will continue to grow, and as the rods are com- FiG. 33 Direction of enamel rods in the gingival third. pleted first just over the tip of the dentine cusp, the con- tinued growth causes an increase in the inclination of the rods in their outer portion. This often leads to a curving of the rods at their outer portion. CHAPTER VII THE RELATION OF THE STRUCTURE TO THE CUTTING OF THE ENAMEL • There are two methods of cutting enamel — to chop or cleave it, or to shave or plane it. Cleaving or Chopping Enamel. — In the cleavage of the enamel the action of the instrument more nearly resembles that of splitting ice than that of splitting wood. The ax for splitting wood is strongly wedge-shaped, and the wedge pries the fibers apart. In splitting ice a small nick is made on the surface and then a sharp blow cracks the ice in the direction of the cleavage. In a similar way the chisel applied to the surface of the enamel makes a slight scratch or bearing on the surface, and the force applied at a slight angle to the direction of the rods cracks the tissue through in the rod direction. The bevel of the instrument is designed to give strength and keenness of edge, not to act as a wedge. In order to cleave the enamel it is always necessary that there be a break or opening in the tissue. Only a small portion can be split off at a time. The edge of the chisel should be placed on the enamel a quarter or half a millimeter from the opening, rarely more, and so piece after piece is split into the cavity. Fig. 34 shows a section of enamel. The edge of the chisel is placed at 1, with the shaft in the relation to enamel rod direction indicated; a tap of a steel mallet will split oft' a piece, and the chisel is moved back to position 2 and a second piece is split off. Undermined enamel will split easily in this way. As soon as a point is reached where the enamel rests on sound dentine, it is recognized by the resistance. Straight enamel can be split off from sound dentine without difficulty if attacked in the proper way, but if the inner portion is gnarled and 74 RE LA TION OF S TR UC T URE TO CUT TING OF EN A MEL twisted, it can only be cleaved by removing the dentine from under it. Such enamel, if resting on dentine, will split Position of chisel in clea\ as far as the rods are straight; but where they begin to twist they will break off, leaving a portion which is very difficult to remove by attacking it from the surface. If the dentine CLEAVING OR CHOPPING ENAMEL 75 is removed from under gnarled enamel, it will crack through in an irregular way, following the general direction of the rods. In preparing teeth for crowns it is often necessary to remove a large amount of enamel. This is always more efficiently accomplished by the intelligent use of sharp instruments than by force. The enamel on axial surfaces, especially in the gingival half of the crown, is usually straight, and if a cleavage line can once be established, the enamel can be more easily and rapidly removed by splitting it off piece after piece than in any other way. In doing this a straight or contra-angled chisel is often the most efficient instrument, and it must be remembered that the "root trimmers" are more properly called "enamel cleavers," and that they are used to cleave the enamel, not to scrape or hoe it off, their form being adapted to give a strong palm grasp of the instrument. Fig. 35 illustrates the use of the enamel cleaver for the removal of gingival enamel from an axial surface. The line of cleavage being established, the edge of the instrument is placed on the enamel half a millimeter from the broken edge, and the force, which should be strong, quick, and sharp, is applied in the direction indicated, and piece after piece is split off, progressing from the occlusal toward the gingival. In preparing the wall of a cavity the outline form should be attained by cleavage, and this is the first step in the preparation of the cavity. After the enamel has been removed by cleavage to the point where the margin is to be laid, the wall must be completed by cutting the enamel in an entirely different way. Planing or Shaving Enamel. — In this manner of cutting enamel the tissue is removed without reference to the rod direction, and without injury to its structure (Figs. 36, 37, and 38). The chisel is used like the blade of a plane. The cutting edge is placed against the surface with the shaft of the instrument almost parallel to it, and the tissue is shaved away. In this way the rods that have been cracked apart 76 RELATION OF STRUCTURE TO CUTTING OF ENAMEL by the cleavage are removed, and the walls arranged in terms of its structural elements so as to gain the required strength of margin. Fig. 35 ^— The use of enamel cleaver in removing enamel. Sharp Instruments. — Chisels and hatchets for use in cleaving or planing enamel must be keenly sharp. If a dull edge is SHARP INSTRUMENTS 77 Fig. 36 Fig. 37 Fig. 38 The use of the chisel in planing or shaving enamel. (Black.) Fig. 39 The relation of the edge of a sharp and a dull chisel. 7S RELA TIOX OF STRUCTURE TO CUTTING OF ENAMEL ])lacc(l on the surface of the enamel it will rest across the ends of many rods, and force applied will only crumble them, but will not split the tissue. The edge must be keen (Fig. .39), so as to engage between the rods and so start the cleavage. Cutting instruments as furnished by dental supply houses are not tempered hard enough to hold an edge. There is no fault to be found with the supply houses for this, for they make them as the dentist wants them, and any dealer will furnish hard-tempered instruments if they are ordered. To use hand instruments successfully in cutting enamel, the stock instruments must either be retempered or the}' must be ordered hard tempered. The cutting edge of Fig. 40 Fig. 41 Fig. 42 The use of the chisel in cleaving enamel. Opening an occlusal cavity. (Black.) the blade of an enamel instrument should be straw-colored when tempered. The chisel and hatchets are the instruments for removing enamel. The burr is the instrument for removing hard dentine. When the burr is used on enamel it should be remembered that it is used as a revolving chisel. It is by the thoughtful use of hand instruments that knowledge of enamel rod direction is gained, and only by the use of them can the enamel walls be pre- pared in terms of their structural elements. In cleaving undermined enamel the edge may be used either with a pulling or a pushing motion. For instance, in opening up a cavity in the occlusal surface of a bicuspid, the buccal SHARP INSTRUMENTS 79 portion of undermined enamel is split off by placing the instrument as shown in Figs. 40 and 41 . The bevel of the blade is held toward the cavity and the shaft of the instrument at a slight angle to the rod direction, and the force is applied in the direction of the shaft. The lingual portion may be removed by placing the instrument as indicated in Fig. 42, the bevel of the blade away from the cavit}^ and the force applied in the direction of the bevel by a pulling force in the direction of the shaft. This is the way in which force is applied on enamel cleavers. The pitch of the bevel in an enamel cleaver and its relation to the shaft of the instrument is extremely important, and the efficiency of an instrument may easily be ruined by careless honing. Every time a cutting instrument is applied to the enamel it must be done with a knowledge of the relation of the cutting edge and the force to the direction of the enamel rods, until it becomes entirely automatic. The author emphatically believes that the acquirement of this knowledge and skill will do more to increase facility and success in the prepara- tion of cavity walls than any other manipulative factor. The preparation of enamel walls requires the continual application of the knowledge of enamel structure. Enamel is a very hard tissue, but it is composed of structural elements, and walls prepared without reference to them will prove their own weakness. CHAPTER VIII THE STRUCTURAL REQUIREMENTS FOR STRONG ENAMEL WALLS From the consideration of the physical character of the enamel, its structural elements and their properties, it is evident that the strength of any enamel wall is dependent upon the arrangement of the rods in the tissue which makes up the walls and their relation to the dentine. Certain requirements for strength can be clearly stated, and these are applicable to all enamel walls. They cannot always be secured with equal facility or perfection, but in proportion as these principles are observed and attained the wall will be strong; as they are imperfectly attained or ignored the wall will be weak and unreliable. When these conditions are understood very many failures can be clearly seen to have been the result of their neglect. Structural Requirements.^ — 1. The enamel must rest upon sound dentine. 2. The rods which form the cavosurface angle must have their inner ends resting upon sound dentine. 3. The rods which form the cavosurface angle must be supported by a portion of enamel in which the inner ends of the rods rest on sound dentine and the outer ends are covered by the filling material. 4. The cavosurface angle^ must be so trimmed or bevelled so that the margin will not be exposed to injury in con- densing the filling material against it (Fig. 43). These requirements should be considered one by one. The Enamel Must Rest upon Sound Dentine. — That is, the enamel plate must have the support of sound dentine, and 1 The cavosurface angle is defined as the angle formed by the surface of the tooth and the wall of the cavity. ENAMEL MUST REST UPON SOUND DENTINE 81 all portions which are undermined by the removal of dentine must be cut away. When the inner ends of the rods which form the enamel plate rest upon sound dentine, the elasticity of the dentine gives to the enamel a certain degree of elas- ticity^, but the enamel itself without this support is extremely brittle. Fig. 44 illustrates these requirements. The enamel plate ABCD rests upon sound dentine. The rods which form FiQ. 43 The structural requirements for a strong enamel wall. the cavosurface angle at B run uninterruptedly to the den- tine, and their inner ends rest on it at E. The rods B, E are also supported by a portion of enamel, ABE, made up of rods whose inner ends rest upon the dentine and whose outer ends are covered in by the filling material, altogether supporting the marginal rods like a buttress. And the cavo- surface angle is bevelled, including from J to y of the enamel 6 82 STRUCTURAL REQUIREMENTS FOR ENAMEL WALLS wall, so as to remove the sharp corner which would be in danger of crumbling in under an instrument. A force that causes it to give way will crack it through its entire thickness. No filling material or substitute for the lost dentine can restore the original condition. An enamel wall should be Fig. 44 The structural requirements for a strong enamel wall: AB, the bevel of the cavosurface angle. The rods forming the margin of the cavity at B reach the dentine at E, and are supported by the portion ABE. considered no stronger after the filling is inserted than it was before. Moreover, when the dentine has been decalcified or destroyed by the action of caries, the acid which has decalcified the dentine has also acted upon the enamel, dis- solving the cementing substance from between the rods, from EX AM EL MUST REST UPON SOUND DENTINE 83 within outward, often to a great extent, and the structure is very imperfect. Enamel that has been so weakened will not withstand the force of mastication, and sooner or later will crack or break away from the filling material. It should be removed and the wall formed in tissue whose structure is perfect. Occasionally cases arise where an Fig. 45 Improperly prepared enamel wall. The portion ABC has the inner ends of the rods cut off and they do not reach the dentine. operator decides to leave some unsupported enamel, but its weakness and the possibility of restoring it if it breaks away without destroying the original operation must always be considered. It is sometimes supposed that it is only neces- sary to have sound enamel resting on sound dentine, but by looking at Figs. 45 and 46 it will be seen that the first require- 84 STRUCTURAL REQUIREMENTS FOR ENAMEL WALLS ment may be present, but not the second. In these illustra- tions the enamel plate is resting on sound dentine, but the tissue has been cut in such a way that the inner ends of the rods have been cut off. The rods that form the cavosurface angle do not extend to the dentine, but run out on the cavity wall at D, and the portion ABC is held together only by the cementing substance. This is not strong enough to B \ Fig. 46 c ^ Improperly prepared enamel -wall. The portion ABC is not supported by dentine. sustain the force necessary to condense the filling material or the forces received upon the surface of the tooth after the filling is completed. It will crack on the line of the cementing substance and chip out. The inclination of the entire wall must be increased to a little more than to reach the rod direction. Such a wall as this may easily be made, in preparing a cavity wall, with a stone or a burr, but would THE RODS FORMING THE CAVOSURFACE ANGLE 85 be unlikely ever to be formed with hand instruments. Such walls as this account for the chipping of many margins and the failure of fillings along the gingival wall. The tissue is cracked to pieces in inserting the filling material, and the pieces fall out later. This occurs often in the gingival walls of compound cavities. Fig. 47 Enamel wall cut in the direction of the rods, supported. It should be trimmed i The marginal rods are not the line indicated. The Rods Forming the Cavosurface Angle Must be Sup- ported. — This is the key to strong enamel walls. The more perfect the support the stronger the wall. If an enamel wall is cut exactly in the direction of the rods, as in Fig. 47, the rods forming the margin are held together only by cementing substance, and a comparatively slight force on the surface in the direction toward the cavitv will break them off. If the 86 STRUCTURAL REQUIREMENTS FOR ENAMEL WALLS same wall is trimmed, as indicated by the line, the same force would do no dama^^e, as the rods which receive it are supported by the portion which is covered by the filling material. Fig. 48 The tip of a worn incisor. The rods forming the angle at A reach the dentine at C, and are supported by the piece ABC. It is interesting to note that in the wearing down of the enamel by use, nature provides the same support for the rods which form the angle of the worn and tooth surfaces. Fig. 48 shows the tip of a worn incisor. The rods at A reach CLASSES OF CAVITIES 87 the dentine at B and are supported by the portion ABC. When caries occurs on an abraided surface it starts by the rods at the dento-enamel junction, chipping out and forming a, protected niche for the lodgement of a colony. Bevel the Cavosurface Angle. — It is not always necessarj' to bevel the cavosurface angle where the rods are inclined toward the cavity. In such places the rods forming the margin are well supported and the angle need not be bevelled unless it is so sharp that it would be in danger of being injured. Fig. 49 The two classes of cavities. Those with the rods inclined toward the cavity, ana those with the rods inclined away from the cavity. There are two reasons for bevelling the cavosurface angle: (1) To protect a sharp angle from injury; (2) to gain support for the marginal rods. The first occurs where the enamel rods are inclined toward the cavity, the second where they are inclined away from the cavity. Classes of Cavities.— From a consideration of the direction of the enamel rods in the tooth crown, and the positions where caries begins on the enamel, enamel walls may be divided, according to their structural type, into two classes (Fig. 49) : 88 STRUCTURAL REQUIREMENTS FOR ENAMEL WALLS 1. Those in which the enamel rods are inchned toward the cavity, characteristic of cavities on occlusal surfaces and cavities beginning in fissures and pits. 2. Those in which the enamel rods are inclined away from the cavity, characteristic of cavities on smooth surfaces. In the firsi class it is comparatively easy to obtain a strong margin, and this is fortunate, for when the filling is completed the margin will be subjected to the full force of mastication. In the second it is comparatively difficult to obtain a strong margin, but only sufficient strength is required to withstand the force of condensing the filling material, a6 after the filling is completed it will be obliged to withstand little force from mastication. From a careful observation of the failures of filhngs (his own and those of other operators), the author believes a very large number are due to structurally imperfect enamel walls. A study of enamel structure as related to cavity preparation will do more to improve the quality of the operation and to increase the facility of its execution than any one factor. This study is a clinical study guided by examination of the microscopic structure of the tissue. In operating at the chair the detail of enamel rod direction as it is applied to cavity preparation is learned, but to do so hand instruments must be used and a sufficient knowledge of the tissue must have been acquired to think of it always in their use in terms of its structural elements. CHAPTER IX THE PREPARATION OF TYPICAL ENAMEL WALLS The steps in the preparation of an enamel wall are: 1. The cleavage of the enamel until the outline form of the cavity is reached. 2. The trimming of the enamel walls. 3. The preparation of the margins. Every enamel wall should be prepared according to these steps. The first not only removes the tissue more or less disintegrated and weakened by caries, but also places the margin of the filling in a position where it is not likely to be covered by the growth of a colony of bacteria. It also determines the direction of the enamel rods so that the walls can be completed in terms of its structural elements. The second step is accomplished by the shaving or planing process, and should always increase the inclination of the entire enamel wall slightly, so as to extend a little beyond the rod directions, and remove the portions that have been cracked or splintered by the cleavage. After cleavage the enamel wall will usually have a more or less whitish look. This is caused by the cracking of the cementing substance between the rods. The light is refracted by the air in these microscopic spaces and imparts this whitish or snowy look to the tissue. These portions are removed by planing or shaving, and the tissue obtains its bluish translucent appearance. The third step is also accomplished by the planing process, and should be carried out with two objects in mind: (1) To so form the cavosurface angle that the tissue will not be liable to injury in the condensation of the filling material against it, and (2) to leave rods whose outer ends will be 90 PREPARATION OF TYPICAL ENAMEL WALLS covered by the filling material to support those which form the actual margin of the cavity. The steps in the })re])aration of enamel walls may be made more clear by photomicrographs. Plate III shows a portion of enamel close to a carious cavity which is to be extended to the left. The chisel is placed close to the margin and the portion is split off. The wall then appears whitish, for, as is seen, the cementing substance has cracked in several places, disturbing the structure, and in several places rods have been broken across. The wall must now be planed so as to increase the inclination of the entire wall slightly, and finally the cavosurface angle must be bevelled, involving from ^ to ^ of the thickness of the enamel wall to give support to the rods forming the margins. In this case the rods are straight and parallel, but in Plate IV they are twisted. If the dentine is removed from under this enamel and the chisel placed as indicated, the portion will be split out, but not only has the tissue been splintered, but a considerable portion is left in which the rods have been broken across. By feeling of the margin with the chisel this can easily be determined, and the angle of the wall must be increased by planing so as to leave the wall in the position shown in Plate IV, 3, and finally the cavosurface angle must be bevelled. Preparation of Simple Occlusal Cavities. — Caries often begins in the mesial and distal pits of the upper bicus- pids, and in preparing the cavities for filling they must be united. Fig. 50 is a buccolingual section through a first superior bicuspid. Suppose caries has reached the dento-enamel junction in both the mesial and distal pits, and the}' are to be united along the groove. A small spear drill is carried into the mesial pit until the dento- enamel junction is reached, then a small inverted cone burr is carried into the dentine just under the enamel and drawn from the dentine to the surface of the enamel. When a narrow cut has been made from the mesial to the distal pit, a chisel placed at the edge of the opening wdll split out the enamel as indicated in Fig. 51. Xow the walls must be planed so as to bring the buccal and lingual walls into the i» It 3 =• 2. - ■^ i ^ (5 GO CD C PI a> ^ o CO 23 3 ■£ C ~ a c :^ ^ S c 2. J. Z- -5 0) — y tri 1 nam rods ]0 (t -' 2 'l -> d •5^3 — ^ ! .^ X " rt *" Q, J in r- _ X S ^ y Z a ■■q_ ^ «■ S I ~> >-^ ^ ~ rS Z z. m 2 i < _ — ^. * 2 X 3 ■s r- 5 S ~ ^ r: 3 25 < 2 0) n. -J * •£ "■ CB ; ^ c ? ££. 3 ^ r rt -• (t r < 0) n (D r* (t a ;l -J ^ 23 r > PLATE IV T 'iF^'-^l l ^ Preparation of Enamel Wall in Gnarled Enamel. 1, enaniel wall as cleaved, showing breaking across rods and slivering at a. 2, wall as sniooth, but not extended to remove short rods whose inner ends are cut off at b. 3, wall extended and trinimed to a position of strength. D, dentine; De, dento-enamel junction; c, cavosurfaee angle; 6, point where inner ends of rods are cut off; a, slivering of the tissue. (About 80 X) PREPARATIOX OF SIMPLE OCCLUSAL CAVITIES 91 axial plane, and the structural requirements will have been completed (P'ig. 52). Fig. 53 shows the relation of the cavity to the crown. Fig. 50 Occlusal Assure in an upper bicuspid, showing direction of rods. (About 80 X) 92 PREPARATION OF TYPICAL ENAMEL WALLS It has often been advised to allow the filling to extend on to the natural slopes of the cusps, as indicated in Fig. 54. Fig. 51 / # The same section as Fig. 51. sbowing the position of the chisel in cleaving the enamel to open the cavity. PREPARATION OF SIMPLE OCCLUSAL CAVITIES 93 Fig. 52 Preparation of enamel walls in occlusal fissure cavities (the same as Figs. 50 and 51). 94 PREPARATION OF TYPICAL ENAMEL WALLS Fig. 53 The relation of the cavity to the crown (the same as Figs. 50 and 51). The trimming of the walls instead of lapping the filling material on the slope of the cusps. PREPARATIOX OF SIMPLE OCCLUSAL CAVITIES 95 It will be seen, however, that a stronger enamel wall and a stronger edge of filling material will be obtained if the Fig. 55 Caries beginning in an occlusal defect of a molar. (About SO X) enamel wall is bevelled to the point where the margin of the filling is desired and the filling finished to this position. 96 PREPARATION OF TYPICAL ENAMEL WALLS Fig. 55 shows a buccolingual section through a molar with a small cavity in a mesial pit. Caries has undermined Fig. 56 The preparation of the enamel walls of the cavity shown in Fig. 55 the enamel slightly toward the buccal, but has attacked the enamel on the surface, extending toward the lingual farther PREPARATION OF SIMPLE OCCLUSAL CAVITIES 97 than the enamel has been undermined at the dento-enamel junction. Applying the chisel to the surface, the undermined enamel is split away, as is indicated in Fig. 56. The buccal wall is planed until it is in the axial plane, and the cavo- surface angle bevelled. It is not necessary to extend the cavity to the lingual beyond the point where sound dentine is reached, but the disintegrated enamel on the surface must be removed. The enamel wall is, therefore, inclined about 6 centigrades lingually from the axial plane, and it is not Fig. 57 ^^\ A The relation nf tl,e cavity to the crown (the same section as shown in Figs. 55 and 56). necessary to bevel the cavosurface angle. The rods are inclined toward the cavity and the rods forming the margins are well supported, and the cavosurface angle is not so sharp as to be endangered in condensing filling material. Fig. 57 shows the relation of the cavity to the crown. All occlusal defects should be filled as soon as the decay has reached the dento-enamel junction, as all progress of the disease beyond that point requires sacrifice of tissue which otherwise would be saved, and the enamel wall becomes less and less strong. Fig. 58 shows a much more exten- 7 98 PREPARATION OF TYPICAL ENAMEL WALLS Fig. 58 A larger cavity in (he occlusal surface of a molar. The position of the chisel in opening the cavity FiQ. 59 The preparation of the cavity shown in Fig .58. PREPARATION OF SIMPLE OCCLUSAL CAVITIES 99 Fig. 60 A gingival third caviti' in a bicuspid, showing the cleavage of the occlusal and i gingival walls as cleaved. _ 100 PREPARATION OF TYPICAL ENAMEL WALLS Fig. 61 The preparatiun of the cavity shown in Fig. 60. GINGIVAL THIRD CAVITIES 101 sive occlusal cavity, one that has been neglected until the enamel has been broken in, and as a result there was much unnecessary loss of tooth structure. The chisel is applied to the surface as indicated, and the undermined enamel broken down until the sound dentine is reached. On the buccal, the enamel wall is cut to the axial plane, and the cavosurface angle bevelled. If the decay in the dentine had reached the tip of the dentine cusp, it would be necessary to remove the tip of the enamel cusp and incline the wall A gingival third cavity in a molar. about 8 centigrades buccally from the axial plane, in order to obtain a strong wall, and then the cusp would be replaced by filling material. On the lingual the undermined enamel is removed, and the wall inclined slightly lingually from the axial plane and the cavosurface angle bevelled a little. Fig. 59 shows the relation of the cavity to the crown. Gingival Third Cavities. — Fig. 60 is a buccolingual section of a superior bicuspid, showing a break in the enamel in Fig. G3 '1. Wall as_cleaved. Fig. 64 2. Wall as trimmed. Preparation of occlusal wall of Fig. 62. (About 70 X) GINGIVAL THIRD CAVITIES 103 the position of a gingival third cavity. The occlusal wall is cleaved to find the enamel rod direction, then planed to Fig. 65 A ca-Wty in the lingual pit of a lateral incisor. The position of the chisel in opening the cavity. 104 PREPARATION OF TYPICAL ENAMEL WALLS increase the inclination slightly, leaving it about 8 centigrades occlusally from the horizontal plane, and the cavosurface angle bevelled to obtain support for the marginal rods. The gingival wall is prepared in the same way, inclined gingivally about G centigrades from the horizontal plane, and the cavosurface angle bevelled. Fig. Gl shows the walls prepared. Fig. 66 The preparation of the gingival wall of the cavity shown in Fig. 65. Fig. 62 is a similar section from a molar. After chopping away the occlusal wall until the cavity has been extended to the point of greatest convexity of the surface, the wall is seen to be in the condition shown in Fig. G3. Near the sur- face some rods have broken across, and near the dento-enamel junction the same thing has happened, but in the rest of the distance the cleavage has followed the enamel rod direction. The inchnation of the wall is increased by planing until this roughness has been removed, and then the cavosurface angle is bevelled to support the marginal rods, and prepara- tion is complete, as shown in Fig. G4. GINGIVAL THIRD CAVITIES 105 Fig, 65 shows a cavity in the Ungual pit of a superior lateral incisor. Caries has undermined the enamel to a Fig. 67 The preparation of the cavity ishown in Fig. 65. lOG PREPARATION OF TYPICAL ENAMEL WALLS \ considerable extent, and the cavity will have to be larger ! than would otherwise have been necessary. Placing the • chisel close to the occlusal margin, as indicated, the enamel i is chij^ped away in that direction and around the circum- | ference. On the lingual wall the chisel may be reversed and used with a pulling motion, like a hoe. In this way i the undermined enamel is chipped away and the tip of 1 the marginal ridge removed. The wall is then planed into 1 the horizontal plane and the cavosurface angle })evelled. \ Fig. (36 shows the structure of the gingival wall, and Fig. 67 the relation to the crown. CHAPTER X STRUCTURAL DEFECTS IN THE ENAMEL The formation of enamel begins at the dento-enamel junction, and the tissue is laid down from within outward, so that the enamel in contact with the dentine is formed first and the surface of the crown last. Enamel formation begins at several points, for each crown, the exact number and position of which has been the subject of much investi- gation. When enamel formation begins, these points are close together, but they are carried farther apart by the growth of the dental papilla, and are not united for some time. The separate enamel caplets unite first at the dento- enamel junction, and as the formation of the thickness of the enamel progresses at these lines of union, there is always more or less disturbance in structure. Even where the union seems perfect, sections will show more or less disturbance of enamel rod direction, arrangement of the rods, and relation to the cementing substance. Every operator and student of dental anatomy is familiar with the developmental lines. On the occlusal surfaces they are usually marked by well-defined grooves, but upon the axial surfaces the grooves may be very slight, scarcely more than slight depressions of the surface, and consequently they are not thought of. It will be found, however, that on these lines there is less perfect enamel structure, and con- sequently the tissue is not as strong, and these lines must be avoided in the preparation of enamel walls. The cause of disturbance of structure will be better understood after study of the development of the tooth germ and the formation of enamel in the chapter on Dental Embryology, but some 108 STRUCTURAL DEFECTS IN THE ENAMEL details of the cause should be touched upon here. The study of the diagrams of the growth of the tooth crown will illustrate the conditions (see Chapter XXVII), and shows a bucco- lingual section through the tooth germ of a bicuspid just before the formation of the dentine and the enamel begins. The odontoblasts (dentine forming cells) and the ameloblasts (enamel forming cells) are in contact at what will be the dento-enamel junction. The odontoblasts form dentine on their outer surface, beginning at the tip of the dentine cusp, and progress from without inward and extend down the slopes of the cusps. The ameloblasts form enamel on their inner surface and progress from within outward and down the slopes of the cusps. In this way little caplets of dentine covered by enamel are formed over the horns of the dental papilla; the caps are, of course, thickest where forma- tion has been going on longest. While these caps are forming, the dental papilla is increasing in size, and so they are carried farther and farther apart (Figs. 68 to 73). As soon as the calcifications reach each other at the dento-enamel junction and unite, the increase in the diameter of the dental papilla ceases. The layer of ameloblasts, which are tall col- umnar cells, now cover the surface of the enamel and receive their nourishment and the materials for the forma- tion of enamel from the blood supply through the stratum intermedium. As the blood supply comes from above, it is evident that the cells high up along the slopes of the cusps will receive most, while those at the bottom of the groove get what is left. The formation is, therefore, more rapid along the slopes and less rapid at the point of union. As growth continues, this difference in supply increases, and accordingly formation at the bottom of the groove is first slowed and finally stopped, and the result is a defect. The taller the cusps the greater will be the interference and the deeper the defective groove. In studying sections (Figs. 74 to 78) it is very noticeable that teeth with long pointed cusps have more open grooves, and the defect often extends almost to the dento-enamel junction. STRUCTURAL DEFECTS IN THE ENAMEL 109 Fig. 68 *5''' « Fig, 69 w Diagram showing the growth of the crown of a bicuspid. 110 STRUCTURAL DEFECTS IN THE ENAMEL Fig. 70 Fig. 71 Diagram showing the growth of the crown of a bicuspid. STRUCTURAL DEFECTS IN THE ENAMEL IJl Fig. 72 Fig. 73 Diagram showing the growth of the crown of a bicuspid. 112 STRUCTURAL DEFECTS IN THE ENAMEL Fig. 74 The sectioii from which Figs. 68 to 73 were drawn: A, tip of dentine cusp, B, lines showing little caps of enamel formed before calcifications from separate centres united; C, lines showing amount of enamel formed when calcifications united. Fig. 75 Occlusal defect from an old tooth. Fig. 76 A deep open groove. Fig. 77 A shallow groove. 114 STRUCTURAL DEFECTS IN THE ENAMEL The bands of Retzius, which are the incremental Hnes of the enamel, should be studied about these grooves. It will be seen that they always dip down around the groove, and that more enamel has been formed between one band (Figs. 84 and 85) and the next on the slope of the cusps than at the bottom of the groove. In teeth with very flat, low cusps the closure of the grooves may be very perfect, leaving only a slight depression (Fig. 77). Fig. 78 A very deep groove, showing the effect of caries at the bottom. The importance of these defects as positions of beginning caries cannot be overestimated, as they furnish ideal con- ditions in areas that would otherwise be immune, and they are the positions in which the attacks of caries are first manifested. These occlusal grooves appear in great variety. STRUCTURAL DEFECTS IN THE ENAMEL 115 Fig. 79 The pit in a lateral incisor filled with coronal cementum. Interglobular spaces are Fig. 80.— Occlusal surface of the lower third molar, showing the grooves. Fig. 81.— The same tooth sliced for sectioning: 1, the piece from which the sec- tion shown in Figs. 82 and 83 was ground. 116 STRUCTURAL DEFECTS IN THE ENAMEL Some are simply shallow open grooves, in which the surface of the enamel is perfect (Fig. 74); some are very deep and entirely empty (Figs. 75, 7G, and 78) ; others are apparently filled with a granular, more or less structureless calcified material which appears to have been deposited in the groove after the enamel was completed (Figs. 79, 84, and 85). This is probably of the nature of cementum. It was formed after the enamel was completed, but while the tooth was enclosed Fig. 82 The section ground from 1, Fig. 81, showing the depth of the fissure. in its follicle in the crypt in the bone. It is to be compared with the coronal cementum that is characteristic of the complex grinding teeth of the ungulates and other herbivor- ous animals. A study of these defects furnishes the basis for the operative rule that "all grooves must be cut out to the point where the margin will be on a smooth surface." For if they are not, a defect will be left at the margin of the cavity which offers ideal conditions for the beginning of a new decay. When caries begins in such a defect at the STRUCTURAL DEFECTS IN THE ENAMEL 117 margin of a filling, it progresses at the bottom of the defect until the dento-enamel junction is reached, and then extends in the dentine and may destroy the entire cro\yn without Fig. S3 Higher magnification of the fissure shown in Fig. 82. (About 60 X) lis STRUCTURAL DEFECTS IN THE ENAMEL showing upon the surface (see Chapter XII). The extent of these defects is much greater than would be supposed from the observation of the teeth in the mouth. Fig. 80 shows the occlusal view of a lower third molar, extracted because of disease of the peridental membrane, from a man aged about forty years. Examining these grooves with a fine-pointed explorer, it would not stick any place. No operator would think of cutting them out and filling them. Fig. 84 An occlusal defect in a worn tooth. The fissure is filled with coronal cementum. The crown w^as sawed through from buccal to lingual, as shown in Fig. 81, and the piece marked 1 is shown in Figs. 82 and 83. The grooves are open two-thirds of the distance to the dento-enamel junction, and show slight action of caries. Suppose caries had started in the central pit, and a small round filUng had been made, open defects would be left at the margin where every groove radi- STRUCTURAL DEFECTS IX THE ENAMEL 119 ated from the central cavity, and these would be just as liable to recurrent decay as it was originally, and, if caries occurred, it would progress at the depth of the groove, reach the dento-enamel junction, and progress in the dentine, Fig. 85 Higher magnification of Fig. 84. The fissure filled with granular calcified material. Notice the direction of the bands of Retzius around the fissure. until the occlusal enamel was so undermined that it would break in under the force of mastication. On the other hand, if the grooves are cut out to a point where the cavity margin 120 STRUCTURAL DEFECTS IN THE ENAMEL will be on a smooth surface, there is no possibility of recur- rent caries if the filling material is properly inserted. This one illustration, which might be duplicated a thousand times, Structural defects in developmental grooves on axial surfaces. ] Fig. 87 B .-;: A A-C "i; V"5-^ .!_ -A Structural defects in developmental grooves on axial surfaces. STRUCTURAL DEFECTS IN THE ENAMEL 121 therefore is the rational basis for the rule, "All grooves must be cut out to their end." Caries does not occur in all open grooves. Fig. 76 shows an open groove in a section from a tooth in which the wear indicates that it was not from a young person, but most of the grooves that escape are not open, but more or less entirely filled with structureless calcified matter or coronal cementum. Figs. SO, S4, and So are very good illustrations of this class of grooves. Fig. 88 Defects on the axial surface in the enamel. The condition in pits from which grooves extend, as the lingual pits of incisors and the buccal pits of molars, show the same condition as the grooves, except that the defect is both broader and deeper. But pits that are sometimes found on the tips of cusps and on smooth surfaces show an entirely different structural condition, and will be considered under atrophy in Chapter XII. 122 STRUCTURAL DEFECTS IN THE ENAMEL In places where the union of the enamel plates seems per- fect, as, for instance, on the labial surface of the incisors or the buccal surface of the bicuspids, and the line of union is marked only by a slight depression of the surface, the section will show disturbance of structure. Fig. 86, a Fig. 89 A section through such a defect as that shown in Fig. 88. (About 80 X ) drawing made by Dr. Black a good many years ago, shows such a position. At the surface the rods and their arrange- ment seem very perfect, but from a point about one-third the distance to the dento-enamel junction there are no STRUCTURAL DEFECTS IN THE ENAMEL 123 rods at all, but apparently a number of calcospherites in a granular calcific substance. In Fig. 87, another of Dr. Black's illustrations, the rods are very irregular, and are separated by large areas of structureless calcified material. Grooves are often found in unusual or atypical positions. Fig. 88 shows a groove running over the mesial marginal ridge and down on the mesial surface. Fig. 89 shows a section through such a defect. Notice the folding of the enamel into the dentine and the disturbance of the rods about the groove and between its base and the dentine. CHAPTER XI SPECIAL AREAS OF WEAKNESS FOR ENAMEL MARGINS There are certain positions which in the perfect crown are areas of great strength, but which, because of the pecu- liar structure of the tissue in these places, become areas of weakness when cavity margins are made in them. The treatment of beginning caries would lead to no failures in these positions, for cavity margins would never be extended into them, except in the treatment of burrowing caries and neglected cases. The extension of caries at the dento-enamel junction often requires the extension of the margin into the area of danger. In considering these areas and in the prepa- ration of cavities, as well as the areas of imperfect structure considered in Chapter X, it is important to place as much emphasis on the necessity of 7iot extendirig cavity margins into the areas of weakness, as on cutting away the dangerous area and leaving the margin in a safe position, when the area cannot be avoided. In considering the relation of the enamel and dentine, and in studying the arrangement of the enamel rod direction in the "architecture" of the tooth crown, it has been pointed out that the dentine cusps and the dentinal marginal ridges are not directly under the corresponding points on the sur- face of the enamel, but are nearer to the axis of the tooth. The areas on the surface of the enamel, from the point directly over the tip of the dentine cusp or ridge to the tip of the enamel cusps or ridges, become areas of weakness when a cavity is extended into them. Fig. 90 is a photomicrograph of a buccolingual section of a superior bicuspid, and Fig. 91 is a higher magnification of the same, made to illustrate the condition. It will be AREAS OF WEAKXESS FOR ENAMEL MARGINS 125 seen that if decay has extended at the dento-enamel junction to the tip of the dentine cusp, and the enamel walls were left in the axial plane, the rods which form the surface of Fig. 90 B A Buccolingual section of upper bicuspid. Enamel is broken from grinding. A to B, area of weakness for enamel margin. (About 20 X) the enamel from the margin of the cavity to the tip of the cusp "are not supported by dentine/' and would be likely to be broken and fall aw^ay, leaving a defect at the margin Fig. 91 Enamel over tip of dentine cusp: D, dentine cusp. (About 80 X) From same section as Fig. 90. AREAS OF WEAKNESS FOR ENAMEL MARGINS 127 of the filling. If decay beginning in the groove or pit has extended only to point C, Fig. 90, the wall may be trimmed in the axial plane and an ideal wall produced; but if it has reached point D, Fig. 90, it must be inclined bucally, so as to remove the tip of the cusp, as indicated in the dotted line, and the cusp restored by the filling material. The region of the surface indicated by A-B, while an- area of Fig. 92 A bicuspid cut for sectioning. Sections were ground from the positions marked by the lines 1, 2, 3, 4 in B, and are sho-svn in Figs. 93, 94, 95, and 96. strength in the perfect tissue, becomes a position of weak- ness when cavity margins are extended into them. A careful observer wiU find many failures that are the result of bad enamel wall preparation in these areas. The same conditions exist in the region of the marginal ridges. Figs. 96 and 97 show the mesial marginal ridge of a superior bicuspid. If this is filled before the destruc- tion of dentine has extended beyond the point A, the mesial wall ma^ be cut in the axial plane as indicated; but if it has reached the tip of the dentine ridge at point J5, it must be inclined mesial ly, so as to reach the tip of the enamel ridge. 128 AREAS OF WEAKNESS FOR ENAMEL MARGINS FiQ. 93 Section ground from 1, Fig. 92, through the mesial oblique ridge. (About 30 X) AREAS OF WEAKNESS FOR ENAMEL MARGINS 129 Fig. 94 Section ground from 2. Fig, 92 (About 30 X), 130 AREAS OF WEAKNESS FOR ENAMEL MARGINS Fig. 95 Section ground from 3, Fig 92, through distal marginal ridge. (About 20 X) AREAS OF WEAKNESS FOR ENAMEL MARGINS 131 Fig. 96 Section ground from Fig. 92, through mesial part and marginal ridge. If caries has extended at the dento-enamel junction to A, the -wall may be in the axial plane; if it has reached B, the wall must be inclined as indicated by the dotted line. (About 30 X) 132 AREAS OF WEAKNESS FOR ENAMEL MARGINS Figs. 98, 99, and 100 show the distal marginal ridge in a sec- ond molar. Notice the inclination of the rods from the tip of the dentine ridge. If decay has reached this point the wall Fig. 97 A higher magnification of Fig. 96, showing enamel rod directions in the region of the marginal ridge. must be inclined distally, so as to reach the rod direction, or a frail margin will be left and one which will not sustain the force of mastication. Neglected caries in the lingual AREAS OF WEAKXESS FOR EXAMEL MARGIXS 133 pits of incisors often present the same conditions. Fig. 65 shows a section through such a pit in a lateral incisor, and Fig. 06 shows the gingival wall. If this were prepared by inclining the gingival wall only slightly, a very frail wall Fig. 9S An upper molar, showing the position of the section shown in Figs. 99 and 100. Fig. 99 The section ground from Pig. 9S. would be left. It should be cut down to the horizontal plane, as indicated, and the marginal ridge restored by the filling material r The same conditions are often encountered in the preparation of simple cavities in the mesial or distal surfaces of incisors, when caries has followed the dento- 134 AREAS OF WEAKNESS FOR ENAMEL MARGINS Fig. 100 The distal marginal ridge, showing the enanael rod direction. (About 30 X) AREAS OF WEAKXESS FOR ENAMEL MARGINS 135 Fig. 101 A B An upper bicuspid, showing the position of the section shown in Fig, 102. Fig. lO: Section from the central piece shown in Fig. 101, showing the dinection of cleavage on the mesial surface and the effect of caries on the tissue. 136 AREAS OF WEAKNESS FOR ENAMEL MARGINS enamel junction toward the lingual. Fig. 1 03 shows a superior central incisor from which sections were cut as indicated. Suppose caries to have begun in the region of the contact point and to have extended to the point a. If the lingual enamel wall were prepared at the line A, Fig. 105, a verj^ frail Fig. 103 A superior central incisor, showing the position of sections in Figs. 104, 105, and 106. Fig. 104 Section 1, Fig. 103, showing the enamel worn from the marginal ridges. wall would result. Force coming upon this wall from the Hngual, by the occlusion of the lower incisors, would be likely to break out or crack a triangular piece of enamel, and the filling would fail along the lingual wall. If, however, the wall be laid in the line at B, a strong wall is produced, against AREAS OF WEAKNESS FOR ENAMEL MARGINS 137 which gold can be properly condensed without danger, and which will withstand the force of occlusion. Dentists are often tempted to prepare simple cavities in the mesial surfaces of first and second bicuspids and occa- sionally in the molars. If this is ever done, it must be with the full knowledge both of the liability of recurrence of caries and the structure of the enamel, for experience shows that Fig. 105 Section 2, Fig, 103, showing position of weak and strong lingual walls. such operations usually fail, either by recurrence of caries at the buccogingival or linguogingival angles, or by the breaking out of the enamel of the marginal ridge. Fig. 107 shows the mesial surface of a superior bicuspid. There was a white spot on the contact point, but no actual cavity, as the enamel rods had not fallen out. A section was ground through this point, and Fig. 108 shows a photomicrograph Fig. 100 • "^ '^'' i^^B m^^ d \ ^^^^^^HK'' ^^^^^^^^^ //^^ ^^^^^^^^^^^^^^^B^M^^^^^Ika A higher magnification of the mesial marginal ridge, shown in Fig. 105. (About 60 X) Fig. 107 Occlusal and mesial views of a superior bicuspid, showing position of section. A beginning caries could be seen on the surface, but it does not show well in the picture. The section from the buccal piece is shown in the following illustrations. AREAS OF WEAKNESS FOR ENAMEL MARGINS 139 of it. The enamel rods have fallen out of the disintegrated area, and the decalcification in the dentine is shown (Fig. 109). If this had been treated as a simple cavity the occlusal wall would have required an inclination of 1 8 centigrades occlusally from the horizontal plane to reach the enamel rod direction. Fig. 108 The section ground froia the buccal piece, 1 There is very little support offered by the dentine for the enamel of the marginal ridge, and the portion over to the occlusal groove would be likely to be broken off by the force of mastication. The conditions of the occlusal wall are better shown in Fig. 110. 140 AREAS OF WEAKNESS FOR ENAMEL MARGINS Any number of illustrations of these conditions might be made, but the subject may be summed up by saying: Fig. 109 The region of the carious spot shown in Fig. 107, showing the disintegrated area of the enamel and the action of acid on the dentine. (About 30 x) AREAS OF WEAKNESS FOR ENAMEL MARGINS 141 Fig. no The enamel over the mesial marscinal ridge to the oblique groove, showing a region of weakness for the occlusal wall of a simple proximal cavity. 142 AREAS OF WEAKNESS FOR ENAMEL MARGINS The surface of the enamel from the point directly over the dentine cusp or ridge to the tip of the enamel cusp or ridge, which is an area of great strength in the perfect crown, is a region of weakness for an enamel wall. It is fully as impor- tant not to extend into this area unnecessarily as to form the wall properly when caries has extended so as to involve it. And when caries of a smooth surface approaches a mar- ginal ridge which receives the force of occlusion, the wall must be extended so that the enamel receives full support from sound dentine. CHAPTER XII THE EFFECT OF CARIES ON THE STRUCTURE OF THE ENAMEL The action of acid upon the enamel has been fully con- sidered in Chapter VIII, and it should be carefully studied before considering the effect of caries on the structure of the enamel, for this cannot be understood unless the relative solubility of the rods and cementing substance and the relationship of the two structural elements are clearly in mind. During the last ten years there has been a great increase in knowledge of the beginning of caries of the enamel and the extent of tissue injury before an actual cavity is pro- duced. This has placed a tremendous emphasis upon the value, for the preservation of the teeth, of the treatment of caries in its early rather than in its later stages. It is safe to say that if caries progresses until a patient is aware of a cavity, the tooth has been injured more than is necessary in the most radical treatment of the same cavity in its beginning stages. One who has not studied carefully the effect of caries on the structure of the enamel, so as to recognize the extent of injury to the structure of the tissue by its appearance to the naked eye, can never be considered fit to prepare cavities as a treatment for the disease. The beginnings of caries must be divided into two classes: (1) Those occurring in natural defects of structure; (2) those beginning upon smooth surfaces. Caries Beginning in Natural Defects of Structure. — These are the positions in which caries first appears and in which it presents the greatest intensity, because they offer ideal conditions. Such open grooves and imperfectly closed pits in the enamel as have been illustrated in Chapter XI become 144 THE EFFECT OF CARIES ON THE ENAMEL filled with food debris, which furnish ideal culture media for acid-forming bacteria. At the opening of the defect the acid is washed away by the sahva as fast as it is formed, but at the bottom of the groove it is confined and acts upon the enamel, dissolving out the cementing substance from between the rods and following the rod direction toward the dento- enamel junction. The form of the disintegrated tissue in such positions is always that of a cone or wedge, with the apex at the opening of the pit or groove and the base toward the dento-enamel junction. The formation of acid in these positions is often so rapid and the confinement so perfect that the carious process here manifests its greatest intensity, the action often dissolving the rods as well as the cementing substance and progressing across the rods. But even when the action follows the rod direction, the form will be broader toward the dentine, as the rods are inclined toward the defect. Figs. Ill and 112 show split teeth illustrating the disintegration of the enamel around occlusal defects. The disintegration area appears white by reflected light because the cementing substance has been removed from between the rods and the resulting air spaces refract the light. As soon as this disintegration reaches the dento-enamel junc- tion, the acid formed passes through the now porous enamel and acts much more rapidly upon the dentine. Because of the branching of the dentinal tubules at the dento-enamel junction, the action upon the dentine spreads rapidl}^ along this line. Soon some of the loosened rods be ween the bottom of the defect and the dentine are either entirely dissolved or displaced or dislodged, and the microorganisms are admitted to the dentine. The decalcified dentine matrix becomes food material for the bacteria, and the space pro- duced by the destruction of tissue furnishes greater space for decomposing foodstufi^s. The acids formed attack the enamel from within outward, producing what has been called backward or secondary decay of enamel. At the mouth of the defect the acid is still washed away, and there is little action upon the tissue. The condition progresses, therefore, until, as in Fig. 113, the entire occlusal enamel THE EFFECT OF CARIES ON THE ENAMEL 145 has been undermined, and all of the undermined area has been greatly weakened by the solution of the cementing sub- Fia. Ill A split tooth, showing caries beginning in an occlusal groove. Fig. 112 A split tooth, showing caries progressing in an occlusal groove. 10 146 THE EFFECT OF CARIES ON THE ENAMEL stance from between the rods. In general sections of such areas as shown in Fig. 116 the disintegrated area appears dark by transmitted Hght. Fig. 114 shows the progress of secondary decay from an occhisal cavity. In this way it often happens that the entire occlusal enamel is destroyed before the original defect is noticeably enlarged. The general form of the disintegrated area in caries beginning in natural defects may be described diagram- maticall}', as in the enamel a cone or wedge with the apex toward the mouth of the defect and the base toward the dento-enamel junction, and in the dentine a cone or wedge with the base at the dento-enamel junction and the apex toward the pulp. Fig. 113 A split tooth, showing the undermining of the occlusal enamel by caries spreading at the dento-enamel junction. Caries Beginning on Smooth Surfaces. — Caries upon smooth surfaces of the enamel is always due to the growth of a colony of bacteria which becomes attached to the surface by the formation of material, causing them to adhere to the surface and at the same time confining their acid products in contact with the enamel preventing its dissipation in the saliva and allowing it to combine with the inorganic CARIES BEGINNING ON SMOOTH SURFACES 147 salts of the tissue elements. This is not the place to con- sider the bacteriology of caries, but the effect upon the structure of the enamel cannot be understood without a Fig. 114 A section shoTving the undermining of the enamel and secondary or backward decay at 1. clear conception of the microbic plaques. A growth of masses of microorganisms upon the surface of a tooth does not constitute a plaque. ]Many very filthy mouths are found where most of the surfaces of the teeth are covered 148 THE EFFECT OF CARIES ON THE ENAMEL by thick, furry masses, and where there is little or no attack of the enamel. Either acid is not formed or it is at once lost by solution in the saliva. Caries shows the greatest intensity in comparatively clean mouths, in which something in the nature of the saliva causes the bacteria to produce a tough zooglea, which attaches them to the tooth surface and con- fines the products of their activity. This zooglea presents some of the phenomena of a dialyzing membrane. Through it the microorganisms receive their food materials, and their products are neutralized by chemical action on the surface upon which the colony is growing. Colonies lodge in the most favorable spots and extend from these points into areas that are less liable to maintain their attachment. The more perfect the confinement of the acid, and the more rapid the rate of its formation, the greater w^ill be the intensity of the destructive process. The more easily the colony is able to maintain itself in its position and extend upon the surface, the greater is the liability. As the colony becomes thickest at the point of beginning, it is evident that the most acid is formed here, and it is therefore the point of greatest intensity. It is also the point at which the growth began, and therefore the spot where the action on the tissue has been longest in operation. It is also apparent that there may be great intensity with limited liability, and great liability with very low intensity, and the effect upon the tissue will be different in the two cases. The appearance of the tissue becomes an index for esti- mating the intensity and liability in a given case. The char- acter of the effect of the disease on the appearance of the enamel, as well as the direction of the extension upon the surface of the tooth, become most important factors in the diagnosis of any case, and the diagnosis is the basis for the treatment required. The increased appreciation of the extent of disintegration of the enamel before an actual cavity is apparent in a tooth has been one of the most important results of Dr. Black's study of caries of the enamel in the last ten years. The author has been intimately associated with this work, and has been amazed at the extent PROGRESS OF CARIES 149 and character of the effect of caries upon the structure of the enamel in what may be called the early stages of the disease. Progress of Caries. — A colony of bacteria becomes attached to the proximal surface of an incisor just to the gingival of the contact point, and remains there some time. If the surface of the tooth can then be examined, a white spot will be seen at Fig. 115; the area appears white because the cementing substance has been removed from between the enamel rods, as will be seen later, and the air that occupies the spaces diffuses the light. If a tooth is split through such a spot and viewed from the surface, the appearance Fig. 115 Fig. 116 A superior central incisor, showing a white spot just to the gingival of the contact point. A split tooth cut through such a white spot as is shown in Fig. 115. will be as shown in Fig. 116. If a section were ground through the spot and the tissue preserved, the ends of the enamel rods would be seen pointed and projecting like the pickets of a fence, giving the same appearance as that produced by the action of acid upon a ground section, as illustrated in Fig. 16, Chapter IV. The surface of the enamel is therefore longer smooth, but roughened. The roughness may no often be felt by passing a very fine pointed steel explorer over the surface. If the colony be dislodged at this stage it is evident that it is much easier for a new one to become attached. These whitened areas are often invisible unless 150 THE EFFECT OF CARIES ON THE ENAMEL the tissue is dried, because the saliva fills the spaces. If the surface is dried the refraction of the light by the air whitens the affected area. A good comparison is furnished in a very familiar phenom- enon. Snow is white because the air and the microscopic ice crystals are of different refracting index, and the light is diffused by passing from air and ice crystals. If a snowball is saturated with water it loses its whiteness and becomes translucent, because the water, which is nearly of the same refracting index as ice, fills the spaces between the ice crystals, and the light is not diffused. If the white area of such a tooth is split through the centre with an aluminum disk charged with emery powder, the enamel rods will be found entirely separated by the solution of the cementing sub- stance, and the cross-striation will be much more apparent because the unevenness in the diameter of the rods has been increased by the action of the acid. Formerly it was impossible to grind a section through such a spot and preserve the tissue. In 1902 the author ground, by the old hand methods, a large number of sections for the study of enamel rod directions. Fig. 107, Chapter XI, shows the mesial surface of a bicuspid split for sectioning. There was a white spot in the region of the contact point that can scarcely be seen in the photograph. When the central section was ground and mounted (Fig. 108, Chapter XI), it was seen that the enamel was disintegrated through its entire thickness and the acid had affected the dentine, and all the enamel rods were lost from the disintegrated area. Until methods were devised by Dr. Black, it was impossible to preserve the tissue and examine its condition. These methods demonstrate definitely that in the disintegrated area the cementing substance is dissolved in large areas before any of the rods are dissolved or destroyed. The first sections of such areas were obtained by polishing the surfaces and cementing the split tooth to the cover-glass with balsam, completing the grinding and mounting without loosening the section. In this way the spaces between the rods were filled with balsam and so were held in place. PROGRESS OF CARIES 151 Fig. 117 shows a photograph of a section made in this way, and the spaces between the rods and the distinct cross-stria- tion are seen. Later it was fonnd that by dehydrating and Fig. 11^ A thin section of carious enamel ground on the cover-glass with balsam: E, sound enamel; X, carious enamel in wliich the cementing substance had been dissolved from between the rods. immersing in a solution of brown shellac, the shellac could be made to take the place of the lost cementing substance, then the polished surface of the sawed-out section could be fastened to the cover-glass with shellac, and the specimen 152 THE EFFECT OF CARIES ON THE ENAMEL handled more easily. Fig. 118 shows a photograph of carious enamel made in this way. The rods are preserved Fig. 118 Carious enamel ground on the cover-glass by the shellac method. In the region X the cementing substance dissolved from between the rods has been replaced by- shellac. r > < 3 (D Q. a 23 > 0! W D' 0) o ^ '^' CQ D (fi (-K ^ :r 0) T CD ''k'b ^ (D (D :r i? a> l"" 0) c .'^ -* -J ^ T »^ 0) 3 3 (T 11 ^ n> -. '^ (h 3 !-<• a a 0) _ V ■-••■Jil r > < a jj » 3 > (/} ^1 0) r, -5 cQ >-! »j if) •^ 8 O CO 23 m — ! 3 r W W o ■J ^. 13 n r+ D :r =< 0) N 0) 0, o ►-» D a a CQ V ^^ (D > STAGES IN THE PROGRESS OF CARIES 153 in place and the dark shellac marks the disintegrated area very clearly. Stages in the Progress of Caries. — The progress of caries on smooth surfaces of the enamel may be divided in three periods, according to its effect upon the structure of the tissue. 1. From the lodgement of the colony until the action reaches the dento-enamel junction. 2. From the reaching of the dento-enamel junction until the rods begin to fall out. 3. After a cavity is produced. First Period. — The form of the disintegrated tissue in the first period is always that of an irregular cone. Its base is on the surface of the enamel, its outline is the boundary of the colony, and the apex is toward the dentine in the direction of the enamel rods from the starting point of the colony. The inner boundary of the area is never even, but shows flame-like extensions toward the dentine in the direction of the rods. This is more marked in some cases than in others, and sometimes suggests that the presence of a colonv on the surface has been intermittent (Plates V, YI, VII). The boundary between the perfect and the disintegrated area is usually marked by a darker area, the significance of which is not now understood. If the disease progresses continuously the affected tissue always appears white by reflected light, but if the progress has been intermittent, especially if there have been considerable periods in which no colony has been attached to the surface, the area darkens, becoming brownish or almost black. This is produced by organic materials filling the space between the enamel rods and decomposing, with the probable formation of sulphides of dark color in the spaces. If immunity to caries is attained before the effect upon the tissue has penetrated to the dento- enamel junction, this will occur, and the spot changes from a white to a brownish or black color. Such spots will be found in some places on most teeth extracted from immune persons. Work of Dr. Miller has indicated that such spots FiQ. 119 A section through a white spot in the first period of attack: X, disintegrated enamel; E, sound enamel; D, dentine. STAGES IN THE PROGRESS OF CARIES 155 are more resistant to the progress of caries than perfect enamel surfaces. At any time during the first period, there- fore, the destruction may be arrested by the coming of Fig. 120 A section through a carious spot in the first period. The attack has apparently been slow and intermittent: X, disintegrated enamel; E, sound enamel; D, dentine. Fig. 121 X i. \ A section through a carious spot in the first period, showing the flame-like projections j toward the dentine: X, disintegrated enamel; E, soimd enamel; D, dentine. | STAGES IN THE PROGRESS OF CARIES 157 immunity, which prevents the attachment of colonies to the tooth surface by the formation of plaques. ' Second Period — This period extends from the time when the action of the acid reaches the dento-enamel junction until the rods are destroyed or fall out. As soon as the solution of the cementing substance reaches the dento- enamel junction at the point of the advancing cone, the solution of the inorganic salts from the dentine matrix begins. It must be remembered that the acid is formed by the microorganisms on the surface of the enamel, and filters through the spaces between the enamel rods. The decalcification of the dentine may be considerable, while the surface of the enamel is still preserved. In this period the swelhng of the surface is always noticeable. This results in increasing the area of the contact and therefore allowing the colony to extend its limits, increasing the extent of the surface attack. This is especially noticeable toward the gingival, and is shown in Plate V, which is, however, shown in the first period of caries. In the disintegrated area in this stage, as well as in the first stage, the diameter of the enamel rods is always considerably reduced and the striation rendered more apparent. In caries of great inten- sity but low liability the reduction in the diameter of the enamel rods is rapid, and they are soon destroyed, while the area of the surface attacked is small (Fig. 122). In caries of low intensity but great liability the diameter of the rods is slowly reduced, while the area of surface attacked, and consequently the area of disintegration, is large (Fig. 123). These conditions should be studied in the macroscopic appearance of caries at the chair. The decalcified dentine matrix shrinks and more or less of a space is formed under the enamel. The action of the acid follows the tubules of the dentine toward the pulp, and spreads through their branches later- ally near the dento-enamel junction so that the form of the disintegrated dentine is always that of a cone, with the base at the dento-enamel junction and the apex toward the pulp chamber. It is important, however, to remember that in 158 THE EFFECT OF CARIES ON THE ENAMEL Fig. 122 A tooth split through a spot, showing great intensity but low liability Fig. 123 A tooth split through spots, showing low intensity but great liability. STAGES IX THE PROGRESS OF CARIES 159 this stage no microorganisms have entered the tissue, and the effect upon it is the result of the action of substances formed upon the surface. The extent of enamel disintegra- tion and decalcification of dentine, in this stage, is much greater than anyone supposed before such specimens as the present illustrations were made. Fig. 124 A drawing showing the microorganisms of caries growing through the dentins tubules. (G. V. Black.) Third Period. — This embraces the period after the enamel rods have begun to fall out and an actual cavity is apparent. As soon as this occurs the surface of the tooth at the point where the formation of the colony began is destroyed and the protected point is lost, and the extension of surface attack ceases. The microorganisms are admitted to the den- tine, where they grow through the dentinal tubules, spreading rapidly at the dento-enamel junction (Fig. 124). The dentine is always decalcified in advance of the penetration of the microorganisms. The acid formed within the cavity attacks 160 THE EFFECT OF CARIES ON THE ENAMEL the cementing substance between the enamel rods, and proceeding from the dento-enamel junction outward. This is called secondary or backward decay of the enamel, and as a result of it, large areas are disintegrated until they are sufficiently weakened to break into the cavity. This con- dition is shown in Fig. 1 14, in which the area indicated by A has had the cementing substance entirely removed from between the rods, and is in the same structural condition as the disintegrated areas in the first and second stage. It is safe to say that in the past few cavities have been filled until the enamel has caved in. It is equally certain that in a large proportion of cases, by the time this has happened, the removal of all disintegrated tissue will require a greater loss of tooth substance than would be required for the prevention of a new surface attack, at the margin of the filling, if the case had been treated as a beginning instead of a burrowing decay. ATROPHY Atrophy is a disturbance in the structure of the enamel caused by an arrest or perversion of the function of the enamel-forming tissue during development. It may be caused by any diseased condition which is serious enough to produce marked disturbance of nutrition, but it is especially liable to follow infectious diseases that affect the epithelium, such as scarlet fever, measles, etc. There are all grades of manifestation of this condition, from a slight disturbance in the perfection of structure to complete loss of a portion of the tissue. In all cases the portion of the tissue which was being formed at the time of the disturbance shows a modifi- cation of structure. The imperfect tissue, therefore, corre- sponds to the bands of Retzius and follows their direction, not the direction of the enamel rods. If, for instance, an atrophied groove shows upon the surface of the enamel at A in Fig. 26, the disturbance in structure will not follow the enamel rods to the dentine, but the band of Retzius, and will reach the dento-enamel junction at B. ATROPHY 161 Character of the Effect on the Tissue. — In atrophy the for- mation of the rods is first affected, the cementing substance becoming greater in amount. Conditions which produce sHght disturbances are marked simply by a prominent band of Retzius. These are iUustrated in Figs. 125 and 126. In such sections the globules of which the rods are composed are more imperfectly fused and the difference in the refracting index between the rod substance and the cementing substance is greater. The cementing substance also seems to contain actual pigment. The zone of dentine which was being formed at the same time usually shows a zone of interglobular spaces, the character of which will vary with the character of the defect in the enamel. Fig. 125 Labial surface of a central incisor, showing atrophy grooves. The stain in the groove makes it appear deeper than it is. Fig. 126 is a part of a section from this tooth. In strongly marked cases it appears that no enamel at all is formed during a longer or shorter period, and when forma- tion begins again the portion that should have been formed is left out and the new formation is telescoped on to the old. When the pathological condition begins to affect the enamel- forming organ, the rods are more and more imperfectly formed and finally disappear, cementing substance contin- uing to be deposited. The entire crown, therefore, is short- ened and has the characteristic stunted appearance. This will be understood by a study of Figs. 127, 128, and 129. On the surface of the tooth, where the new formation joins 11 162 THE EFFECT OF CARIES ON THE ENAMEL the old, there is a groove the depth of which is determined partly by the duration of the disturbance and partly by its severity. The grooves of atrophy are usually accompanied to a greater or less extent by pits in which the formation of rods seems to have failed entirely and the tissue is composed of an imperfect granular material representing defective inter- prismatic substance. Fig. 126 injurj' from atrophy. The growth of enamel -was interrupted, but not stopped. The zone of interglobular spaces in the dentine separates the dentine formed before the interruption from that formed after. (Black.) White Spots. — White spots are often seen (Fig. 130) in the enamel of one or two teeth. The surface in these cases is usually smooth and vitreous like the rest of the tooth. Sections ground through such spots show that in the area of the spot the rods have been perfectly formed, but no cement- ATROPHY 163 ing substance. When such an area is entirely within the substance of the enamel, as they usually are, and the surface of the tooth is covered with normal tissue, the spot remains white, or with the same appearance it had when the tooth erupted. If the defective structure reaches the surface they may become more and more stained. Fig. 127 Section of incisors, showing two zones of atrophy, appearing as two grooves on the surface. (About 8 X) (Black.) The spaces between the rods are occupied by air and the refraction diffuses the light, causing the white appearance. There is no way to remove these spots except by polishing away the tissue, and this is never advisable. When the white 164 THE EFFECT OF CARIES ON THE ENAMEL area reaches the surface of the enamel it is not smooth and poHshed, as in the normal condition, but rough and chalky. One or two cases have been reported in which all of the Fig. 128 Atrophy: A portion showing the incisal groove of Fig. 127 more highly magnified. The dark Une separates the enamel of the first formation from that of the second. Notice that the second is lapped on the first. The narrow line of interglobular spaces is seen in the dentine. (Black.) Fig. 129 The second, or gingival, groove shown in Fig. 127. The overlapping of the third formation on the second is not as great, but the discoloration is greater. The band of interglobular spaces in the dentine is much wider, (Black.) ATROPHY 165 enamel of all the teeth was of this character.^ Instead of the normal color the tissue was originally white like a sheet of paper, but it became very much stained and dis- colored. It was soft and chalky and could be picked to pieces with an instrument. When ground sections were examined it was found that the interprismatic substance was entirely absent and that the rods were standing unsup- ported with spaces between them (Fig. 131). Fig. 130 A section through a white spot in the enamel. In the white area the enamel rods are without cementing substance between them. (Black.) Mottled Enamel. — In certain restricted geographical areas there seems to be a tendency to imperfections in the forma- tion of the enamel. In these places the teeth of many children present when they erupt white or mottled areas. There may be only a few spots on a few teeth, or it may 1 Black's Operative Dentistry, vol. i, p. 35. 166 THE EFFECT OF CARIES ON THE ENAMEL involve all of the enamel of all the teeth. In some places a large proportion of all the children born and brought up in the district have teeth more or less disfigured. These mottled teeth seem to be often accompanied by greatly freckled skin. Fig. 131 ;^/ Enamel from a h^ection ground by Dr. Black trom near the cusps of one of the teeth in Dr. Prunty's case, showing enamel rods breaking into bundles, which end in spiculse. This enamel has no cementing substance between the rods. Its color was white like paper. (Black.) Nothing is now known as to the cause of this condition or how the enamel organ is affected to produce this result. The study of this enamel has shown that the enamel rods are perfectly formed, but that the cementing substance is entirely absent in the mottled area. When the spots are white the spaces between the rods are entirely empty; when they are brown or a dark color, they are filled more or less with some sort of coloring matter. In many cases there is more or less pigment in these spaces before the teeth erupt, and in some cases they grow darker with age. CHAPTER XIII THE DENTINE The dentine may be defined as a connective tissue whose intercellular substance is calcified. It is apparently homo- geneous in structure, but penetrated by minute canals, which contain protoplasmic projections from cells lying within a cavity enclosed by the tissue. The Function of the Dentine. — The dentine makes up the mass of the tooth, giving to it its general form, each cusp and root being indicated in it. It gives to the tooth its elastic strength, and the enamel, being hard and very resistant to abrasion, but extremely brittle, is dependent upon the elastic support of the dentine. This has been elaborated to a considerable extent in the chapter on the Dental Tissues. The fact that the dentine gives the strength to the tooth should never be lost sight of in operating, and sound dentine should never be sacrificed unnecessarily in the preparation of cavities. Structural Elements of the Dentine.— The structural ele- ments of the dentine may be stated as: 1. The dentine matrix. 2. The sheaths of Newman and the dentinal tubules. 3. The contents of the dentinal tubules or the dentinal fibrils. While these are the elements of which the tissue is com- posed, there are other characteristic appearances found in the dentine, caused by special conditions or arrangement of these elements which must be studied. These are the granu- lar layer of Tomes, the interglobular spaces and the lines of Schreger, and secondary dentine. Origin of the Tissue (Histogenesis). — The dentine, like all of the other calcified tissues except the enamel, is a 168 THE DENTINE connective tissue, and is formed by the dental papilla, which is a conical papilla of connective tissue rich in bloodvessels and covered on its surface by the layer of dentine forming cells, the odontoblasts. The dentine is formed from without inward, leaving the remains of the dental papilla in the cavity of the formed dentine as the dental pulp. Before the tooth is erupted, and up to the time that the full length of the root is formed, a characteristic thickness of dentine is formed, which is called the primary dentirie. After this time dentine is formed by the pulp only intermittently, in response to irritations and trophic impulses, producing secondary dentine. Secondary dentine is always more irregular in the arrangement of the tubules, and more imperfect in structure than the primary dentine. The boundary line between two periods of dentine formation can always be picked out by changes in the direction or character of the dentinal tubules. The Dentine Matrix. — The dentine matrix is a solid, appar- ently homogeneous, and very elastic substance, through which the dentinal tubules extend. It is translucent in appearance and slightly yellowish in color. In broken or split sections to the unaided eye it has a yellowish color by reflected light, and a characteristic luster due to the refraction of light by the tubules. In ground sections, by transmitted light, under the microscope, it is very translucent and shows no indication of structure. The matrix consists of an organic basis of ultimately fibrous character, yielding gelatin on boiling, with which the inorganic salts are chemically combined. The relation of organic and inorganic matter in the dentine matrix is similar to the condition in the bone matrix and that of all calcified connective tissues. Apparently the organic basis is first formed, and then the inorganic salts are combined with it in a weak chemical union. If the dentine is treated with dilute acid, the inorganic matter is dissolved and the organic basis is left retaining the form of the tissue. If the organic matter is burned out, it leaves the inorganic matter in the character- istic form. THE SHEATHS OF NEWMAN 169 Von Bibra gives the following analysis of perfectly dry dentine : Organic matter 37.61 Fat 0.40 Calcium phosphate and fluoride 66 . 72 Calcium carbonate 3.36 Magnesium phosphate 1.08 Other salts 0.83 Dr. Charles Tomes pointed out that such analysis as this failed to take account of about 8 per cent, of water which is held as water of combination, and which is driven off at about red heat. It is evident that the organic matter in the dentine is of two kinds — the organic basis of the matrix, which is of gelatin yielding character, and the protoplasmic contents of the dentinal tubules. Variations, therefore, in the pro- portion of organic and inorganic matter in the dentine might be caused by differences in the proportions of organic and inorganic constituents of the matrix, or by variations in the size of the tubules and the amount of material contained in them. If dentine changes in its degree of calcification with age, this might be brought about by the reduction in the size of the tubules, or by the adding of inorganic constituents to the matrix. The ultimately fibrous character of the dentine matrix can be made out only in various stages of decalcification and decomposition. In the original condition no trace of the fibrous character can be seen. By maceration with acids and alkalies the intertubular material assumes a fibrous appearance, as if bundles of white connective-tissue fibers had been fused together. There is apparently no definite arrangement of these fibers and there is no indica- tion of the arrangement of the substance in layers. The Sheaths of Newman. — There has been much discussion as to i:he character of these structures, which were first discovered in 1863 by Newman. Some investigators have denied their existence entirely, explaining the appearance 170 THE DENTINE in some other way. These structures are in no sense a sheath surrounding tlie dentinal fibril and lying in the dentinal tubule, but are that portion of the matrix which forms the immediate wall of the tubule. That this material differs from that which occupies the rest of the space between the tubules is certain, and is shown by the examination of ground sections, the action of stains upon ground sections, and the action of the matrix when boiled with strong acids Fig. 132 ""-•-- . --^ ^■^Jtrmi., M ' ^fsp >* ^-~---— ^ ™ ■j 1 ^i f c' Dt D Dentine showing tubules in cross-section: Dt, dentinal tubules; D, dentine matrix; S, shadow of sheaths of Neumann. (About 1150 x) and alkalies. In Fig. 132, a photograph of a ground section, there is evidently a difference in the refracting index of the portion of the matrix immediately surrounding the tubules. Apparently the sheaths of Newman are composed of a material similar to that forming elastic connective-tissue fibers, and known as elastin. This substance is very resistant to the action of acids and alkalies. After the remainder of the intertubular material has been destroyed by boiling DIRECTION OF TUBULES IN CROWN PORTION 171 with strong acid, the sheaths remain Uke hollow elastic fibers, having the appearance of pipestems, which resist long continued action of the boiling acid. Some authors have suggested that the great elasticity of the dentine was largely due to the presence of this substance. The Dentinal Tubules. — The dentine matrix is penetrated everywhere by minute branching tubules, which radiate from the central cavity or pulp chamber and extend to the outer surface of the dentine at the dento-enamel junction or the dentocemental junction, where they end blindly or in irregular enlargements. These tubules are from 1.1 to 3 microns in diameter. One hundred measurements^ made at random from ground sections gave the extreme measure- ment: 3, largest; 1.5, smallest; and average, 2.95. Fifty measurements from one longitudinal section of tubules at their pulpal extremity gave an average of 2.6; largest, 3; smallest, 1.5; and 50 measurements at the dento-enamel junction of the same section gave the following: Average, 1.2; largest, 1.5; smallest, 0.75. These measurements were made with an eye-piece micrometer, using yV oil immersion objective and No. 3 ocular. At the present time there is a fertile field for investigation offered in regard to the size of dentinal tubules. ]\Iany statements have been made that have not been supported by tabulated measurements, and no definite statement can be made as to the variations and size of the dentinal tubules in different teeth, the teeth of different animals, or in the human teeth at different ages. Direction of Tubules in Crown Portion. — In the crown portion and the gingival portion of the dentine the tubules pass from the pulp chamber to the dento-enamel junction, or the dentocemental junction, in sweeping curves, which were called by Tomes the primary curvatures. These have been described as /- or *S-shaped (Fig. 133). The tubule tends to enter the pulp chamber at right angles to the surface, and to end at the dento-enamel junction at ' KoUiker gives 5 microns, also Schafer; Owen, 2.5 microns. 172 THE DENTINE right angles to that surface. In the dentine forming the axial walls of the pulp chamber the tubules make two bends in passing from the pulp chamber to the surface of the dentine. In the first the convexity is directed apically, in the second it is directed occlusally. The outer extremity of the tubule is, therefore, considerably farther to the occlusal than the point at which it opens into the pulp chamber Fig. 133 A section showing tlie primarj- curvatures of the dentinal tubules in the crown portion. (About 20 X) (Fig. 134). The outer part of this double curve is often complex instead of simple (Fig. 135). The course of the dentinal tubules is not a direct one, but that of an open spiral. This may easily be demionstrated by changing the focus up and down in examining sections cut at right angles to the direction of the tubules. When examined in longitudinal sections this spiral course gives to the tubule DIRECTION OF TUBULES IN CROWN PORTION 173 the appearance of having httle wavy curves throughout its length. These have often been called the secondary curva- tures. Each wave represents a turn in the spiral. As many as two hundred have been counted in the length of a single tubule, or about one hundred in a millimeter of length. Fig. 134 A section showing the primary curvature of the dentinal tubules in the gingival portion. (About 20 X ) The dentinal tubules give off minute lateral branches, which extend from one tubule to another. These are very minute, and in the crown portion of the dentine are not at all conspicuous, but in the region of the dento-enamel 174 THE DENTINE junction the tubules branch dichotomously, each fork having about the same diameter as the original tubule (Fig. 136.) These forkings of the tubules resemble the appearance of the delta of a river on the map. The branches anastomose with each other very freely. This anastomosis of the tubules at the dento-enamel junction is very important in determining the spreading of caries in this area. It probably also explains the sensitiveness of this area noticed in the preparation of cavities, which will be noted again in considering the sensitiveness of the dentine. Fig. 135 A section showing compound curves near the dento-enamel junction. (About 80 X ) ' The Dentinal Tubules in the Root Portion.^In the root portion of the dentine the tubules ordinarily show only the secondary curves, their general direction being at right angles to the axis of the pulp canal. Throughout their course they give off an enormous number of very fine branches extending from tubule to tubule. These are so numerous that in suitably prepared sections they may be said to look like the interlacing twigs of a thicket or the rootlets of plants in the soil. Fig. 137 gives a very good idea of the appearance. At the dentocemental junction the tubules end in irregular DENTINAL TUBULES IN THE ROOT PORTION 175 anastomosing spaces, which cause the appearance of the granular layer of Tomes (Fig. 138). From a consideration of the preceding it will be seen that it is usually not difficult to determine whether a field of Fig. 136 Dentine at dento-enamel junction, showing tubules cut longitudinally: Dt, dentinal tubules; D, dentine matrix. (About 760 X) dentine seen under the microscope was taken from the crown or the root of a tooth. The structural characteristics of the two regions may be summarized as follows: In the croicn, the tubules show both the primary and the secondary curves. 176 THE DENTINE In the root, the tubules show only the secondary curves. In the croum, the lateral branches are few and inconspicuous and the tubules branch in a characteristic way at the dento- enamel junction. In the root, the lateral branches are very Fig. 137 Dentine from the root, showing tubules cut longitudinally. (About 700 X ) numerous throughout the length of the tubule, and they end in the characteristic spaces of the granular layer of Tomes. The Dentinal Fibrils. — In life the dentinal tubules are occupied by protoplasmic projections of the odontoblasts THE DENTINAL FIBniLS 177 known as the dentinal fibrils, or fibers of Tomes. x\s the dentine matrix is formed and calcified under the influence of the odontoblasts, a portion of their protoplasm is left in the tubules of the matrix as the dentinal fibril. These Fig. 138 Granular layer of Tomes: L, lacunae of cementum; Gt, granular layer of Tomes; Ig, interglobular spaces. (About 200 X) structures were first described by Jolm Tomes, who recog- nized tKeir true character. They may be demonstrated in decalcified sections, and they will be seen projecting from the odontoblasts, when the pulp is removed from a freshly 12 178 THE DENTINE extracted tootli, by cracking it and picking the pulp out. In tliis way a portion of the fibril is pulled out of the tubules. The fibrils will be considered more especially in connection with the pulp, to which they properly belong. The Granular Layer of Tomes. — The granular laj^er of Tomes is the outer layer of the dentine next to the cementum. The granular appearance is caused by irregular spaces in the dentine matrix which connect with the ends of the dentinal tubules, and wdiich are filled with protoplasm continuous with that of the fibrils. Tomes first called attention to this layer, and for this reason it bears his name. With magnifications of from 50 to 100 diameter it is easily seen in ground sections either longitudinal or transverse, and appears as a layer filled with little dark spots or granules, the spaces which have been filled with the debris of grinding. It is separated from the cementum by a thin clear layer, apparently of structureless dentine matrix, which is more apparent in higher magnifications. The granular layer is sometimes seen in the crown portion just under the enamel, but it is never as well marked in this position. The layer is seen in sections ground from freshly extracted teeth as w^ell as from old dry teeth, showing that these are true spaces and are not produced by the shrinkage of par- tially calcified dentine matrix. Tomes called the spaces in the granular layer "interglobular spaces," but this term should not be used, as the structures generally known as the interglobular spaces are different in location and char- acter, and will be considered later. The granular layer is not seen in decalcified sections. So far as the author is aware, no one has called attention to this fact before. In decalcified sections stained with hematoxylin and eosin the position of the granular layer is always occu- pied by a clear layer which takes the stain in an entirely different way from the rest of the dentine matrix, and in which no indication of spaces can be seen. While the fibrils in the tubules through most of the dentine take the hematoxylin stain and can be easily seen, they cannot be THE INTERGLOBULAR SPACES 179 followed into this clear layer, and no indication of proto- plasmic contents of irregular spaces can be seen.^ Most authorities state that the spaces of the granular layer communicate with the canaliculi of the cementum, as well as with the tubules of the dentine. This the author has been unable to confirm. On the other hand, the granular layer seems to be separated from the cementum by a thin layer of dentine which is clear and apparently structureless. This is separated from the cementum by a dark line, and the first layer of cementum usually does not contain any lacunae or canaliculi. This is supported by some of the experiments that have been made with extracted teeth. In experimenting on the diffusion of drugs through dentine, it was found that liquids sealed in the pulp chambers of extracted teeth could not be detected in the liquids in which the teeth were placed unless the cementum was removed from them. In the recent experiments of Dr. Southwell, of Milwaukee, in which air was forced through the dentine from the pulp chamber, to test the sealing of cavities by filling materials, the air did not escape from the cementum, which would be the case if dentinal tubules connected with the canaliculi of the cementum. If the spaces of the granular layer are filled with the pro- toplasmic enlargements of the ends of the dentinal fibrils, this would give a very reasonable explanation of the sensi- tiveness of slight caries and erosion at the gingival line, as the anastomosis through the granular layer would affect the fibrils of the entire root. The Interglobular Spaces. — There has been considerable misunderstanding in dental histology in regard to these spaces, owing to the confusing of two entirely different things. Tomes called the spaces of the granular layer, which have already been described, interglobular spaces. As has been seen, they are true spaces in the dentine matrix 1 The appearance of the tissue in decalcified sections has led to some doubt in the writer's mind as to the interpretation of the character of the laj-er by authors who have described it. ISO THE DENTINE which connect with the dentinal tubules and are filled with protoplasm. In 1850 J. Czermak^ described areas of imperfectly cal- cified dentine matrix, which appear as spaces in dried dentine, and called them interglobular spaces. These have been so called by most writers since. It seems important to the author that the term be restricted to these and some others used to indicate the spaces of the granular layer, which are of entirely different character. Fig. 139 A drawing showing a zone of interglobular spaces in the dentine. (Black.) The interglobular spaces of Czermak are caused by some disturbance in the calcification of the organic matrix of the dentine. They occur in zones (Fig. 1.39) which correspond to the dentine matrix, being calcified at a given time, and there is usually seen a corresponding disturbance in the calcifica- Beitrag zur Mikro-Anatomie der Menschlichen-Zahne. THE INTERGLOBULAR SPACES 181 tion of the enamel, which was being formed at the same time and manifested as a more or less strongly marked atrophy band. Fig. 140 ^im^'^m^- ^^ ^-^'^^^^ r^^r^^ Interglobular spaces in dentine. (About 60 X ) Fig. 141 Interglobular spaces in dentine. Some emptj', some filled wiiL JlLtis. Clbout SO >, ' In the calcification of the dentine matrix the inorganic salts are combined with the organic matrix in spherical areas which become united. The boundaries of these areas of uncalcified matrix are therefore very irregular, and made 182 THE DENTINE up of concave facets where they join the spherical surfaces of the fully calcified matrix (Figs. 140 and 141). A study of the illustrations and the appearance of the layer of forming dentine next to the dental papilla of a developing tooth will make this intelligible. Fig. W2 Interglobular spaces in dentine: Ig, first line of interglobular spaces; Ig', second line of interglobular spaces. (About 30 X ) If the dentine is dried the organic matrix in these areas gives up water and shrinks, and the interglobular spaces become true spaces, partially filled with the shrunken matrix. In this condition they can be filled with colored collodion THE INTERGLOBULAR SPACES 183 or any other material. If, however, they are studied in sections of teeth which have never been allowed to dry, no space appears, and the dentinal tubules continue without change of course or diameter through the area. While they are, therefore, not empty spaces, they are areas of the organic matrix of the dentine which are bounded by globular sur- faces of the fully calcified matrix, and their name is properly significant. Fig. 143 A root broken on a line of interglobular spaces. This tooth was extracted by Dr. G. v. Black, and was pulled apart in extraction, a shows the form of the root and a b the separation on the line of growth. (Black.) Zones of interglobular spaces may occur at any portion of the dentine, either in the crown or root, but they are more common in the crown and near the enamel. Often more than one zone can be seen, as in Fig. 142, which shows two dis- turbances in calcification, and disturbances in the structure of the enamel will be seen at corresponding positions. The zones of interglobular spaces appear in all grades, from a complete band of uncalcified matrix to widely scattered patches. Fig. 143 shows a tooth in Dr. Black's collection which was broken in extraction, because of the presence of such a zone in the root. 184 THE DENTINE The interglobular spaces are of great importance in modi- fying the direction of the progress of caries in the dentine. The Lines of Schreger. — As in the case of the interglobular spaces, there seems to be considerable misunderstanding in the literature, and certain structures which have very different meanings have been called the "lines of Schreger." An arrest in the formation of dentine often occurs before the crown is completed. When the activity has begun again the dentinal tubules follow a slightly different direction. In a longitudinal section this change in the direction of the tubules produces a line. Several such lines may be seen in a single section, though they are by no means to be found in all longitudinal sections. Schreger's lines have been most often confused with zones of interglobular spaces, and they seem to be identical with the incremental lines in the dentine described by Owen. It is unfortunate that these names should have been used, for a thoughtful study of the tissue makes their interpreta- tion perfectly evident, and they are of no great significance. Secondary Dentine.— It is by no means easy to define secondary dentine or to pick out any particular piece of dentine in a section and to say whether it is primary or secondary. In general, the tubules are smaller, fewer, and less regularly arranged in secondary than in primary dentine. In general, it seems that the smaller the remainder of the dental papillae becomes, the more imperfect dentine it forms, until finally it simply throAvs down granular calcified material. The formation of dentine begins at the dento-enamel junction, at a number of points in each tooth, and progresses from without inward (strange to say, exactly the opposite statement has been made several times in papers by very prominent men). This matter will be taken up more in detail in Chapters on Dental Embryology and Dentition. It is enough to say here that in studying all sections of dentine, whether cut longitudinally or transversely, the formation of dentine began at the dento-enamel junction and the dentocemental junction, and progressed toward the pulp chamber. SECONDARY DENTINE 185 From the study of longitudinal and transverse sections it is apparent that a certain typical amount of dentine is formed before the tooth is erupted and while it is coming into full occlusion. This is primary dentine. In it the tubules are very regular in size and arrangement. From Fig. 144 P o^-*^«i^ M^AVl '•X-^it\»\uMiyiM: Secondary dentine: A, margin of pnnuay LkiiUuc .-Lowiiiy u few ul thu tubules continuing into secondary dentine; P, pulp chamber. (About 80 X) this time on the formation of dentine is intermittent, and apparenth' is the response to some outside condition. These conditions may arise in the tooth in which the formation occurs, or the irritation of one tooth may cause tissue forma- tion in all or part of the others. It has not been determined 186 THE DENTINE whether such reflex trophic stimuH are confined to the same lateral half or the same nerve distribution. Apparently the formation of dentine proceeds again, after a pause, in all teeth. It will seem, therefore, that the mere exposure of the entire crown to conditions of thermo-change produces sufficient stimulus to the pulp tissue to cause a renewal of dentine formation. After the first period of rest the dentine Fig. 145 A transverse section of a root, showing the reduction in the size of the pulp and formation of secondary dentine. formed in the second period is so nearly identical, and the direction of the tubules so nearly the same, that it is usually impossible to recognize the junction except at a few points in the circumference of a transverse section. After each period of rest, however, the difference in structure between the succeeding portions becomes more marked. Fig. 144 SECONDARY DENTINE 187 shows an area from a longitudinal section when the line A was the pulpal wall of the dentine. There was probably a considerable period of rest, when for some reason a new formation of dentine was begun. But apparently only some of the odontoblasts took part in the new formation of A transverse section of a root, showing changes in the form of the pulp canal by the formation of secondary dentine. dentine matrix, for not nearly all of the tubules are con- tinued, and those that do continue show a sharp change in their direction and a difference in diameter and character (Figs. 145 and 146). These characteristic changes in the structure of the dentine that is formed as the pulp becomes smaller seem to the author of great practical importance. CHAPTER XIV THE CEMENTUM The cementum may be defined as a connective tissue whose intercellular substance is calcified and arranged in layers (lamellse) around the circumference of a tooth root, the cells being found in spaces (lacunae) irregularly placed in or between the layers. Structurally the cementum is more closely related to the subperiosteal bone than any other tissue, the only differences being that in general the lacunse in bone are much more uniform in size, shape, arrangement of the canaliculi, and their position with reference to the lamellse than those in cementum. In bone the lacunae are usually found between the lamellae. In cementum the lacunae may be between the lamellae, but they are more often enclosed within their sub- stance and they are found most often where the lamellae are thick. Some writers have described Haversian canals in the cementum, but the author has never seen anything that could properly be called an Haversian canal in the cementum from human teeth. Canals containing bloodvessels are not uncommon, but in these the lamellae are never arranged concentrically around the canal, as they are in Haversian systems. For the last fifteen years the author has had under personal observation, in the course of class work, not less than 200 longitudinal sections, and 300 transverse sections of the root, ground from human teeth, and in that time he has never seen what could be called an Haversian canal. In the same time he has examined many hundreds of sections cut through the decalcified jaws of various mam- mals, including the sheep, pig, cat, and dog, with the same negative result. HISTOGENESIS 189 Function. — The function of the cementum is to attach to the tooth the connective-tissue fibers which hold it in posi- tion and support the surrounding tissues. The formation of cementum begins as soon as the tooth begins to erupt, and continues, at least intermittently, as long as the tooth remains in place, whether it contains a live pulp or not. The function of the cementum cannot be too strongly emphasized, and must be continually borne in mind. If, for any reason, the tissues are detached from the surface of the root, they can only be reattached by the formation of a new layer of cementum on the surface of the root, which will embed the surrounding connective-tissue fibers. In order to accomplish this the tissues must lie in physiologic con- tact with the surface of the root, and the connective-tissue cells must be actively functional. That the tissues may be reattached to the surface of a root is both theoretically possible and clinically demonstrable, but for it to occur, biological laws must be observed and the conditions are very difficult to control, especially with the old methods involving the excessive use of strong antiseptics. It is well to remember ''that a dentist can never cure a suppurating pocket along the side of a tooth root," but if the conditions can be controlled the cells of the tissue may form a new layer of cementum, reattaching the tissues and so close the pocket. It is a biological problem, not a matter of drugs, except as they are a means of producing cellular reaction. In view of its function, therefore, the cementum becomes not the least but the most important of the dental tissues, for no matter how perfect the crown may be, without firm attachment the tooth becomes useless and is soon lost. Histogenesis. — The cementum is formed by connective- tissue cells lying between the fibers of the tissue which clothes the surface of the root and which becomes specialized for this function. Their origin is undoubtedly similar to that of the osteoblasts, but they are not osteoblasts, either mor- phologically or functionally, as will be seen later in the 190 THE CEMENTUM study of the peridental membrane, where the cementoblasts and the formation of cementum will be considered. Structural Elements. — The structural elements of the cementum are: 1. The lamellae. 2. The lacunse and canaHculi. 3. The cement corpuscles. 4. The embedded fibers of the peridental membrane. The Lamellae of the Cementum and Their Arrangement. — The lamellae of the cementum resemble those of bone, but they are very much more irregular both in thickness and appear- ance. They may be extremely thin and almost transparent, or they may be quite thick and coarsely granular. They are not nearly as easily observed as those of bone, for in bone the lamellae are marked off by the lacunae which lie between them, while in cementum the lacunae may be entirely absent, and when present are irregularly placed. In the gingival portion of the root the lamellae are always thin and very transparent, and lacunae are seldom seen. The entire thickness of the tissue is transparent, and the appear- ance of the lamellae can be seen only by using a very small diaphragm or oblique illumination. In this position the tissue is largely made up of embedded connective-tissue fibers, which are, however, so perfectly calcified that they cannot be demonstrated in ground sections. In decalci- fied sections they are very easily seen. The cementum becomes gradually thicker in the middle third of the root, and is thickest in the apical third. It will be seen that this increase in thickness is caused chiefly by the greater thickness of each individual lamella. In longi- tudinal sections the cementum is often found becoming suddenly thicker at a certain point, and if examined closely, it will be seen that each layer is continued apically, but with greater thickness. Fig. 149 illustrates this condition near the apex of the root. From a study of the lamellae, therefore, it is apparent that the entire root is clothed with successive layers, and that these layers are formed intermittently, but continue to be formed as long as the tooth is in position. THE LAMELLA OF THE CEMENTUM 191 In a general way the number of layers is an index to the age of the person at the time the tooth was extracted (Figs. 150 and 151). The rate of formation is not uniform; for Fig. 147 ''■:■''> .^ ^^ / kv\iiivv\\\\\vM\ \\\\n\Vu\\\\\VV\\wa\ ^^UiV Hjpertrophy of the cementum on the side of the root of a lower molar near the neck of the tooth. From a lengthwise section, man: a, dentine; h, cementum; c, fibers of peridental membrane. From 6 to c the cementum is normal and the incre- mental lines fairly regular, but at d one of the lamellae is greatly thickened. At e this lamella is seen to be about equal in thickness with the others. The next two lamellae are thin over the greatest prominence, but one is much thickened at g, and both at h. These latter seem to partially fill the valleys which were occasioned by the first irregular growth. (1 in. obj.) Fig. 148 WMfMMI&^^^MMiM^ Hypertrophy from root of cuspid, man, in which the irregularity is confined to the first lamella: o, dentine: 6, thickened first lamella: c, subsequent lamellae, which are seen to be fairly regular. (1 in. obj.) 192 THE CEMENTUM instance, a number of layers may be formed within a short time, and again, a considerable time may elapse between the formation of one layer and the next. The time, however, does not seem to determine the thickness of the layer. Fig. 149 Apex of root of an upper bicuspid tooth with irregularly developed cementum: a, a, dentine; b, b, pulp canals. The lamellse of cementum are marked 1, 2, 3, 4, 5, 6, 7, 8, 9; d, d, d, absorption areas that have been refilled with cementum. It will be seen that the apices of the roots were originally separate, but became fused with the deposit of the second lamella of cementum, and that in this the irregular growth began and was most pronounced. It has continued through the subsequent lamellse, but in less degree. It will also be noticed that the absorption areas, d, d, d, have proceeded from certain lamellse. That between the roots has broken through the first lamella and penetrated the dentine, and has been filled with the deposit of a second lamella. Other of the absorptions have proceeded from lamellse which can be readily made out. The small points, e, seem to have been filled with the deposit of the last layer of the cementum, while others have one, two, or more layers covering them. (2 in. obj.) If a considerable number of teeth of persons of twenty years of age were sectioned, the lamellse counted, and this number compared with the number found in teeth extracted THE LAMELLA OF THE CEMEXTUM 193 from persons of forty, a fairly regular increase in the number of layers will be noticed, and so on, for fifty, sixty, seventy, or eighty years. It is important to remember in connection with this formation of cementum that the teeth move, more or less, under the influence of natural forces throughout life, and that every slight change in position must be accomplished Fig. 150 A transverse section of a root extracted from a young person. The cementum is thin, but is thicker in the grooves on the proximal sides. by the formation of a new layer of cementum, to reattach connective-tissue fibers in new positions or adjust them to new directions of strain. The first layer of cementum is formed while the tooth is still in its crypt, but apparently no connective-tissue fibers are calcified into it. This forms the first apparently clear and structureless layer which lies next to the granular 13 194 THE CEMENTUM layer of Tomes (Fig. 138). Even in the teeth the entire length of whose roots are formed before they begin to erupt, there is no attachment until some stress comes upon the crown. The tooth is lying loose in its crypt and can be picked out with very little force. Bicuspids are often acci- dentally extracted in the extraction of temporary molars. x\s soon as the tooth comes through the gum a new layer Fig. 151 A transverse section of a root from an old person. This root had carried for many years. The section was cracked and one edge broken. of cementum is formed over the entire root, attaching the fibers to its surface, and as the tooth moves occlusally, layer after layer is formed. This will be considered again in connection wdth the peridental membrane. The Lacunse and Canaliculi. — The lacunse of the cementum correspond with the lacunse of bone. They difter from those of bone, however, in that they are more irregular in shape, size, position, and relation to the lamellae, and in the number THE CEMENT CORPUSCLES 195 and direction of the canaliculi radiating from them. In bone the lacunae are fairly regular in shape, the long diameter exceeding the short diameter by about one-third. Sections cut through their long axis give an oval outline, the length of which is about three times as great as the width. Sections cut through their short axis give an oval outline, the long diameter being about twice that of the short. The spaces are, therefore, flattened between the lamellae. In cementum there is no regularity whatever, either in size or in shape. Some are a little larger than the lacunse in bone, some are very much smaller. They may be almost exactl}^ the shape of typical bone lacunae or they may be distorted into almost any form, sometimes being almost stellate, often pear- shaped, sometimes round, and occasionally pyramidal. The lacunae of bone are fairly uniformly placed, and lie between one lamella and the next.^ There is no regularity in the relation of the lacunae of the cementum to the lamellae. They sometimes lie between one lamella and the next, but they are more often entirely in the substance of one. They occur only where the lamellae are thick, and there may be large areas with considerable aggregate thickness of cementum in which there are no lacunae at all. The number and direction of the canaliculi which radiate from the lacunae of cementum is extremely irregular, but in general there are more extending from the lacunae toward the surface than toward the dentine. The Cement Corpuscles. — The cement corpuscles corre- spond exactly to bone corpuscles. They are the cells found in the lacunae. These are simply embedded cementoblasts and are typical connective-tissue cells. They are made up of granular protoplasm and contain a faintly staining nucleus. Extensions of the protoplasm undoubtedly extend into the canaliculi. These cells bear the same relation to the matrix of the cementum that bone corpuscles do to that ' This is not absolutely correct, there being much more irregularity in the arrange- ment of the lacunae in thick subperiosteal bone than in either cancellous or Haversian system bone. To be strictly accurate, the above statement must be limited to Haver- sian system bone (Plate X). 196 THE CEMENTUM of bone. What this is is not known in any definite way, but it is known that when bone corpuscles are killed or die, the matrix becomes a foreign body, and is either absorbed or cut off from the portion in which the corpuscles are living, to be absorbed or cast out as a sequestrum. The same conditions are true of cementum. P'or instance, there are many cement corpuscles in the lacuna in the region of the apex of the root. If this portion be bathed in pus for a long time, the cement corpuscles are killed, and the tissue becomes saturated with poisonous materials, so that tissue cells cannot lie in contact with it and live. In order to restore a healthy condition, the necrosed cementum must be removed mechanically until tissue is reached with which cells may lie in physiological contact without injury. Conditions which can only be understood through a knowledge of the structure of the tissue often arise in connection with the treatment of alveolar abscess. It should always be remem- bered that the treatment of an abscess is a biological problem. The Embedded Fibers of the Peridental Membrane. — The embedded fibers of the peridental membrane are in the strictest sense comparable with the fibers of Sharpe in bone. They are, however, in many places much more perfectly calcified. To appreciate the relation of the embedded fibers to the matrix, the tissue must be studied both in ground and decalcified sections. For instance, in the gingival portion, from the study of ground sections, the presence of embedded fibers would never be suspected, but if decal- cified sections are studied it will be found to be almost entirely composed of calcified fibers. In the middle and apical thirds of the root, w^here the lamellse are thicker, the calcification of these fibers is often not as perfect as that of the rest of the matrix. In the preparation of ground sec- tions, therefore, the imperfectly calcified fibers shrink and consequently appear as canals in the cementum. In fact, they have often been mistaken for canals. They are usually not seen unless the section happens to cut in their direction. These will be seen in many of the illustrations of cementum. In Fig. 152 several layers are seen next to the dentine, in EMBEDDED FIBERS OF PERIDENTAL MEMBRANE 197 which no fibers appear, then in several layers the fibers are plainly seen, and finally, the surface layers show no fibers. This probably means that before and after these layers were formed there was a change in the position of the tooth and the fibers were all cut off in this area and reattached in a new Fig. 152 Cementum near the apex of the root: Gt, granular layer of Tomes; L, lacunae; b, point at which fibers were cut off and reattached. (About 54 X) direction, adapting them to the new directions of strain. It is often necessary to study ground sections very closely to determine whether certain appearances are embedded fibers or canaliculi radiating from the lacunae. The appear- ance of these fibers should be studied in Fig. 153. It should be noted that wherever special stress is exerted upon a 198 THE CEMENTUM bundle of fibers the cementum is thick around them. This may be seen in decalcified sections in Figs. 220, 248 and Plate Fig. 153 Two fields of cementum showing penetrating fibers: Gt. granular layer of Tomes; C, cementum not showing fibers; F, penetrating fibers. (About 54 X) XIV and in ground sections in Figs. 152 and 153. When the next layer is formed, if the fibers are cut off, the additional thickness of the last layer is removed. The unequal thick- EMBEDDED FIBERS OF PERIDENTAL MEMBRANE 199 ness of the last formed layer is not seen in the layers beneath it to as great an extent. Fig. 154 Record in the calcified tissue of an absorption repaired: D, dentine; Cm, cementum filling absorption cavity. (About 40 X) Fig. 155 Thick lamellae of cementum with many lacunae, filling an absorption in dentine: L, lacunae; H, Howship's lacunae filled; D, dentine. (About 250 X) 200 THE CEMENTUM Absorption and Repair of the Cementum. — From what has ah-eady been said about the cementum, it will be understood that this tissue is continually undergoing changes, that new layers are being added, and that often before an addition is made there is absorption enough of it at least to cut off the fibers. When an absorption occurs on the side of a root which cuts into the dentine, the excavation in the dentine may be filled by the dentine subsequently formed (Figs. 154 and 155). From a study of ground sections in class work such absorptions are not uncommon. They probably occur when the cusps first come into occlusion in eruption. CHAPTER XV DENTAL PULP Definition. — The dental pulp may be defined as the connec- tive tissue occupying the central cavity of the dentine. It is composed of embr^^onal connective tissue which is more closely related to the tissue occupying the spaces of cancellous bone than to any other. Functions. — The functions of the dental pulp are: 1. A vital function, the formation of dentine. 2. A sensory function responding to thermal change and chemical and traumatic irritation. J^ital Function. — The vital function is the formation of dentine and is performed by the layer of odontoblasts. These cells also, by means of their dentinal fibrils, maintain the same relation to the dentine matrix that the bone and cement corpuscles bear to the matrix of bone and cementum. When the pulp is removed from a tooth its dentine becomes dead dentine in the same sense that bone in which the bone corpuscles have been killed is necrosed bone. That there is a constant reaction between the protoplasm of the odonto- blasts and the substance of the dentine matrix, or that the presence of the living protoplasm is necessary to prevent degeneration of the matrix, is evidenced by the changes in the physical properties of the dentine after the pulp has been lost. That the tooth remains functional after the loss of the pulp is due to the fact that, except at the minute foramina, the dentine is not in physiologic contact with any tissue excepting enamel and cementum, and that the cementum attaches the tooth to the surrounding tissues, receiving its nourishment from the surface and not from the dentine. 202 DENTAL PULP When the pulp is removed and its place filled by a non- irritating material, the dentine becomes entirely encased in cementum, the foramina probably being covered over as the subsequent lamellae are formed. The author wishes to emphasize, however, the vital relations of the pulp to the dentine matrix. Dead dentine is never as good as living dentine, consequently a tooth from which the pulp has been removed can never be considered just as good as one with the living pulp. The production of the dentine matrix is, of course, the principal part of the vital function of the pulp. It is begun in the development of the tooth before the dental papilla is converted into the dental pulp, by being enclosed in the dentine formed. After the tooth is fully formed the pulp retains its ability to build dentine matrix as long as it retains vitality, but this function is exercised only in response to conditions of environment which are probably excited through the intervention of its sensory function responding to thermal change and chemical irritation. The sensory function causes a trophic impulse which is manifested by the production of another portion of dentine matrix reducing the size of the pulp chamber. That this is a reflex and not purely a local matter is indicated by the fact that formations of dentine occur in one tooth when the irritation is in another, and apparently the irritation of one tooth will excite dentine formation in all of the teeth on that side, at least in some instances. On the other hand, purely local responses are found where a few odontoblasts respond to the irritation of their fibrils by the formation of dentine. This matter has been referred to under the heading of secondary dentine, and it is best studied by the record it leaves in the formed tissue. The Sensory Function. — In regard to sensation, the pulp resembles an internal organ, as in its normal condition it is always enclosed in the cavity of the dentine. It has no sense of touch or localization, and responds to stimuli only by sensations of pain. The pain is usually located correctly with reference to the median line, but apart from that it is STRUCTURAL ELEMENTS 203 located only as it is referred to some known lesion. If several pulps were exposed on the same side of the mouth, and in teeth of both the upper and lower arches, so that they could be irritated without impressions reaching the peri- dental membrane, if the patient were blindfolded it would be impossible for him to tell which of the pulps was touched. This characteristic becomes extremely important in diag- nosis. The pain originating from a tooth pulp may be referred to the wrong tooth or to almost any point on the same side supplied by the fifth cranial nerve. The dental pulp is especially sensitive to changes in temperature, amounting almost to a temperature sense, having no exact parallel in any other tissue of the body. This does not amount to a recognition of heat or cold as such, but a special resentment to sudden changes. For instance, if a tooth is isolated and so protected by non- conductors that the soft tissues cannot be stimulated, and a jet of hot and then cold water be thrown upon its crown, it will respond to each with a sharp sensation of pain, but the patient cannot tell which is hot and which is cold. It is the sudden change that produces the reaction. This is the basis of very important differential diagnoses, for, us is true with most organs, in pathologic conditions its sensory function is exaggerated. Histogenesis. — The dental pulp is the remains of the dental papilla. The dental papillae for the temporary teeth appear- ing in the mesodermal tissue of the jaw arches very early in fetal life. The cellular elements are at first very closely placed and large, but they grow smaller and take on the typical form of pulp cells as the intercellular substance is increased. By the sixteenth week the dental papillae for the temporary teeth are covered by a layer of tall columnar cells, which will begin the formation of dentine about that time. After the beginning of dentine formation the trans- ition from the dental papillae to the dental pulp is very gradual, and it would be impossible to draw any sharp line of demarcation between them. 204 DENTAL PULP Structural Elements. — The structural elements of the den- tal pulp are: 1. Odontoblasts. 2. Connective-tissue cells. 3. Intercellular substance. 4. Bloodvessels. 5. Nerves. The Odontoblasts. — The odontoblasts are tall columnar cells which form the outer layer of the pulp adjacent to the dentine, and from which cytoplasmic fibrils extend into the dentinal tubules. Fig. 156 Odontoblasts and forming dentine: E, forming enamel; D, forming dentine; O, odontoblasts; Dp, body of dental papilla. (From photomicrograph by Rose.) The character of the odontoblasts changes very greatly with the age of the tissue, and the activity of dentine forma- tion. While the primary dentine is being formed they are tall columnar cells, each containing a large oval nucleus, THE ODONTOBLASTS 205 rich in chromatin and located in the pulpal third of the cell. From the dentinal end of the cell cytoplasm is con- tinued, without any line of demarcation, into the dentinal tubule as the dentinal fibril. In some instances two fibrils may be sent from a single odontoblast. The character of the odontoblast is beautifully seen in Fig. 156, a photograph by Professor Rose. Fig. 157 r t*:* T % „«l ^ts> g^ j^'i^ ..^^M Odontoblasts. The section cuts obliquely through the odontoblasts: F, fibrils; A^, nuclei of odontoblasts; A'', nuclei of connective-tissue cells; W, lajer of Weil, not well shown. (About SO X) After the tooth is erupted, but while the formation of dentine is actively going on, the odontoblasts, while some- what smaller, retain the same typical appearance. They may be easily demonstrated either in decalcified sections or by removing pulps from the pulp chamber of freshly extracted teeth. Professor Salter has described two sets of processes besides (Fig. 157) the dentinal fibril process. As a result of teasing the fresh pulps, he considered that fine projections of W 206 DENTAL PULP Fig. 158 Diagram of odonto- blasts and dentinal fibrils. (C.H.Stowell.) the protoplasm extended from the sides of the cells, uniting them to the adjoining odontoblasts (Fig. 158). These he called the lateral processes. He also described cytoplasmic projections from the pulpal end of the odontoblasts into the laj'er of Weil. It is probable that these appear- ances were the result of teasing, and are not true structural characteristics, as the work of other investigators has not con- firmed their presence. It is easy to under- stand how teasing the cells apart might produce appearances which might be inter- preted as processes, but careful work upon sections does not show their presence. In old pulps where the formation of dentine has been intermittent and very infrequent for a long time, the odonto- blasts are smaller, lose their columnar form more or less, and become pear-shaped or globular. As dentine is one of the most highly specialized connective tissues, the odonto- blasts are among the most highly differen- tiated connective-tissue cells. They are the only connective-tissue cells of colum- nar form. Morphologically, they are very similar to columnar epithelium, but epi- thelial cells never have such processes as the dentinal fibrils. Occasionally, in young and actively growing bone, osteoblasts are found which are distinctly columnar in form, but they are never as tall as the odontoblasts, and the nucleus is more nearly in the centre of the cell. In the case of the osteoblast the cytoplasmic processes which extend into the canaliculi correspond to the dentinal fibril process of the odontoblast. ' The homologies be- tween the osteoblasts and the odonto- THE MEMBRANA EBORIS 207 blasts have often been lost sight of in the discussions over the character of the latter and their relation to the forma- tion and sensitiveness of the dentine. The Membrana Eboris. — The odontoblasts form a single layer of cells on the surface of the pulp in contact with the dentine. This layer was very early recognized to be related to the formation of the dentine, and was called the memhrana eboris, or the membrane of the ivory. The name has no importance now except as it is found in the literature. Size of the Odontoblasts. — From what has been said it will be recognized that the size and shape of the odontoblasts vary greatly in different sections. This is true not only of pulps from different animals, and pulps at different periods of development, but of different parts of the same pulp. In the coronal portion of a pulp from a fully developed tooth, but one in which the formation of dentine is still going on, the average measurements would be about 5/^ in diameter and 25 to 30/^- in height. During early stages of dentine formation, before the crown is fully formed, they are con- siderably larger and taller, and in the pulps of a calf they are much larger than in smaller animals and man. In a con- stricted pulp, as, for instance, in the mesial root of a lower first molar, the odontoblasts on the constricted sides will be shorter and relatively thicker than on the buccal and lingual, where the long axis of the cell is in the direction of the long diameter of the pulp, but this simply means that the formation of dentine on the constricted side is relatively farther advanced than on the buccal and lingual, and the cells show older phases. It is evident that the supply of nourishment to the cells in the constricted portions is more imperfect, and that the ones farthest from the main vessels are most affected, so that dentine formation is slowed and made more imperfect here, while it still continues in full vigor around the expanded portions of the pulp. This has been spoken of in connection with the study of the dentine (see Figs. 145 and 146). Origin of the Odontoblasts. — The odontoblasts are special- ized connective-tissue cells. It is therefore to be expected 208 DENTAL PULP that they should be formed from undifferentiated connective- tissue cells as osteoblasts are formed from similar cells of the inner laj^er of the periosteum. The odontoblasts are there- fore developed from embryonal cells deeper in the pulp which take their place in the odontoblastic layer. This probably explains the appearance of some sections, and also, the author believes, the views of some men in regard to the odontoblasts and the dentinal fibrils. In some sections from old pulps the odontoblasts seem to be in an incomplete layer, and their form is more like that of typical connective-tissue cells. Connective-tissue Cells. — The cells in the dental pulp, aside from the odontoblasts, are typical connective-tissue cells such as are found in embryonal tissue. They are of three forms — round, spindle-shaped, and stellate. In the crown or bulbous portion the cells are mostly stellate, while in the root portion they are largely spindle-shaped, with the axis of the spindle parallel with the canal. It seems difficult for students to get an idea of their arrangement, and the nucleus is often mistaken for the entire cell. The cells do not lie in contact in a compact tissue, but are widely scattered in the intercellular substance. There is a small ovoid nucleus, which takes the stain deeply, surrounded by a mass of granular protoplasm stretching away into very fine threads. In the spindle-shaped cells the protoplasm is stretched out in only two directions. In the stellate cells there may be three, four, or more, stretching away in any direction. Plate VIII was very carefully drawn with the camera lucida so as to represent accurately the number, size, and position of the cells in that field as seen with the yV oil immersion. It is very difficult in a drawing to represent the third dimension of space, and to show that some of the processes are extending in a plane at right angles to the paper. An idea of this can only be obtained by the very careful use of the fine adjust- ment while studying the cells with the high power. The round cells are probably white blood corpuscles or undifferentiated connective-tissue cells which may develop either into stellate or spindle-shaped. PLATE VII . ft* ^ * v^ A Field from the Coronal Portion of the Pulp froni a Hunian Molar. In the corner the stage micrometer shows y^o of ^ niilHmeter drawn vvith the same^lens. The field sho^A^s the branching of a bloodvessel and the connective-tissue cells of the pialp. Drawn from. J^ oil-ininiersion lens with camera lucida. (About 12,000 X) THE BLOODVESSELS 209 The Arrangement of the Cells. — Immediately beneath the layer of odontoblasts, for a space about one-half or two-thirds as wide as the odontoblastic layer, the cells are very scarce, making a clear line in many sections. This is known as the layer of Weil, and contains many fine nerve fibers which are not stained by ordinary methods. Beyond the layer of Weil for a space perhaps twice as wide as the height of the odontoblasts, the cells are very closely placed. Through the remainder of the pulp they are much more widely but comparatively evenly scattered. The Intercellular Substance. — Very little is really known about the character of the intercellular substance of the pulp. It contains few fibers, and these in no way resemble bundles of white or elastic connective tissue. The appear- ance in the section is more as if a structureless gelatinous material had been coagulated by the reagents. There are, of course, connective-tissue fibers in connection with the walls of the larger bloodvessels and nerves, and to a certain extent in the gelatinous material. In studying the intercellular substance in the sections it is necessary to remember that it is filled with the protoplasmic projections from the cells, and these are stained, appearing like fibers in the matrix. There is need for further investigation of the character of the intercellular substance. The Bloodvessles. — The dental pulp is an extremely vas- cular tissue, and the arrangement of the vessels, the structure of their walls, and the nature of the intercellular substance through which they run render the tissue especially sus- ceptible to the pathological conditions which are associated with alterations in the circulation. Usually several arterial vessels enter the pulp through foramina in the region of the apex. These vessels have their origin in the rich vascular network of the cancellous bone (Chapter on Peridental Membrane). The arteries follow the central portion of the pulp, giving off many branches as they pass occlusally, and finally form a very rich plexus of capillaries near the surface of the pulp. From these capil- laries the blood is collected into the veins, which follow 14 210 DENTAL PULP Fig. 159 *h I A section through the apex of a root showing three foraminse, A, B, and C. THE BLOODVESSELS 211 courses parallel to the arteries, leaving the pulp through the same foramina in the region of the apex. It is important to notice that an artery is entering and a vein leaving the tissue Fig. 160 ..• \ Diagram of the bloodvessels of the pulp. (Stowell.) through very minute canals in the calcified dentine (Fig. 159). Dr. Stowell has made a very beautiful diagram of the arrangement of the bloodvessels in a single-rooted tooth, which is shown in Fig. 160. Preparations such as would 212 DENTAL PULP reproduce this diagram can be made by injecting the blood- vessels with an inert material and destroying the soft tissues by artificial digestion. Structure. — The delicacy of the walls of the bloodvessels is one of the most striking histologic characteristics of the dental pulp. The largest arteries show only a few muscle fibers in the media and a very slight condensation of fibrous tissue for an adventitia. There is no distinct boundary between the capillaries and the veins, and the vessels con- tinue to have only a wall of endothelial cells after they have reached a size much greater than that of capillaries. Because of this peculiarity of structure the statement is to be found in many text-books of histology that the largest capillaries in the body are found in the dental pulp. These vessels should probably not be considered as capillaries, but as veins whose walls have the structure of capillaries. Even in the largest veins the media is very imperfect, and there is only a slight condensation of fibrous tissue to represent the adventitia. This peculiarity of the bloodvessel walls in the pulp renders the tissue peculiarly susceptible to hyperemia and inflammation. Fig. 161 is a photograph of a bloodvessel whose size can be estimated from the number of red corpuscles seen in it, and the wall is made up of a single layer of endothelial cells. There is no indication of either media or adventitia. The intercellular substance of the pulp being of gelatinous, semifluid character, gives no support to these delicate walls. In Plate VIII the author has drawn very carefully, with the camera lucida, using a yV immersion lens, a field showing the branching of a small bloodvessel. The size of the endo- thelial cells, position of their nuclei in the wall of the vessel, and the size, position, and shape of the connective-tissue cells, are represented as accurately as possible. The field is from the coronal portion of the pulp of a human molar. The caliber of such a vessel as this would depend almost entirely upon the blood pressure. The endothelial cells will stretch to a very considerable extent under increased pressure, becoming very thin at all points except around the nucleus. When THE BLOODVESSELS 213 the pressure is decreased the contractiHty of the protoplasm pulls the cells together, making it thicker and less in diameter. It is very important to remember these facts in connection with hyperemia of the dental pulp. It is difficult in such Fig. 161 A pulp bloodvessel, showing the thin wall: C, blood corpuscles in the vessel; Bl, bloodvessel wall showing nuclei of endothelial cells; N, nuclei of connective- tissue cells in the bodj- of the pulp; I, intercellular substance, showing a few fibers. (About 200 X) an illustration to give any representation of the third dimen- sion of space, which is essential to a real understanding of the connective-tissue cells of the pulp. These are bits of cytoplasm with a nucleus forming a small irregular central 214 DENTAL PULP mass, from which the cytoplasm is stretched away in all directions through the intercellular substance, ending in very fine threads. Plate IX is drawn in the same way from a transverse section of the pulp of an unerupted tooth of a sheep. The vessels are all cut transversely and are seen crowded with red blood corpuscles. They are not distended, and some show slight condensation of fibrous tissue around them. In a normal pulp there are many capillaries so small that a single corpuscle passes them with difficulty, but in patho- logic conditions they become distended to many times their normal diameter. All investigators have agreed in finding no lymphatic vessels in the pulp. This is also an important fact in connection with pathologic conditions. The Nerves of the Dental Pulp. — Few subjects in connection with dental histology have received more attention than the distribution of the nerves of the dental pulp, especially in relation to the sensitiveness of the dentine. Support for almost any idea can be found in the literature, but many of the conditions described have been shown to be errors in microscopic interpretation, and many others have failed to receive support by reinvestigation. The most recent work upon this subject was done ten or twelve years ago by Prof. Carl Huber, of Ann Arbor. The author has repeated some of his work, and has never seen any specimen that was contradictory to his statements. Usually three or four nerve trunks enter the dental pulp through the foramina. These contain from eight or ten to thirty or forty medullated nerve fibers. They pass occlusally through the central portion of the pulp, but almost immediately begin to give off branches, which pass toward the periphery, branching and anastomosing in their course. Most of the fibers lose their medullary sheath very soon after leaving the nerve trunk, proceeding as beaded fibers, made up of an axis cylinder with nuclei scattered along it. A bundle of such fibers, breaking up to be distributed to one horn of the pulp, is shown in Fig. 162. Other fibers retain their medullary sheath, following an independent course through the pulp PLATE IX U. \. ^ % '•r r\* .'J /•••' ''.^. • ♦ • • ^ « • ^'i 3.-<^ W w ... # ,^ V > A Field froni the Pulp of an Unerupted Tooth of a Sheep. The bloodvessels are cut transversely. (About lOOO X) THE NERVES OF THE DENTAL PULP 215 tissue, until they reach the layer of Weil, where the sheath is lost and they join the plexus of beaded fibers Wmg in this posi- tion (Fig. 1G3). From the plexus in the layer of Weil beaded fibers are given off, passing between and around the odonto- blasts, forming a network around each cell, and even passing over on to the end of the cell between it and the dentine, but they have never been followed into the dentinal tubules. In no instance and by no method that he has employed, has Dr. Huber been able to demonstrate nerve fibers in the dentinal tubules. Fig. 162 x^^^^ Nerve fibers in pulp from a human mol (About 500 X) The sensitiveness of the dentine, in view of these obser- vations, is due to the presence of living fibrils, connected with living odontoblasts which are in physiologic connec- tion with nerve fibers. It is interesting to note that this is the only instance in which a connective-tissue cell is inter- mediate between the outside world and the nerve fiber. In 216 DENTAL PULP all other instances an epithelial cell is intermediate between the environment and the nervous system. The sensitiveness of the dentine is therefore due to the irritability of the cyto- plasm of the fibril, transmitted through the continuity of cytoplasm to the odontoblasts and their reaction upon the surrounding nerve fibers. The irritation to the fibril may be either traumatic, chemical, or thermal. For instance, salt is sprinkled on exposed living dentine, and a sharp sensa- tion of pain is the result. It may be supposed that chemical Fig. 163 A h •■ B u ■ C H D A ,| 1 j \ I:]:-: I Rose's diagram of nerves and bloodvessels of the pulp. changes are set up in the cytoplasm of the fibril which excite changes in the cytoplasm of the odontoblasts. These react upon the cytoplasm of the nerve fiber, and so are transmitted to the nerve centre, being recognized, in consciousness, as a sensation of pain. In the same way traumatic irritation caused, for instance, by the cutting of dentine with a steel instrument sets up changes in the fibril in the same fashion. It is impossible to conceive of any vital activity of cyto- plasm otherwise than as a form of chemical action or molec- ular or atomic movement of its substance. THE NERVES OF THE DENTAL PULP 217 Certain clinical facts are well explained by these structural facts. It is often noted in the preparation of cavities that the dentine is most sensitive at the dento-enamel junction. This would be expected when it is recalled that at the dento- enamel junction the dentinal tubules fork and the fibrils anastomose, so that an irritation to a few fibrils is not simply transmitted to their odontoblasts and the nerve endings in contact with them, but to all the fibrils, and so to the nerves in contact "^ith all of the odontoblasts. The presence of dilute acids render the cytoplasm of the fibrils much more irritable. The dentine in a carious condition is, therefore, much more sensitive than that in a sound or normal area. The sensitiveness of extremely hypersensitive dentine can often be greatly reduced, if not entirely over- come, by cleansing the cavity thoroughly, washing with tepid water followed by a dilute alkali, drying and sealing for a few days, when it will be usually found that excavation can be carried out without excessive pain. The sealing must be perfect. If it is leaky the cavity will be more sensitive than ever at the end of the delay. Teeth in which the size of the pulp chamber has been reduced by the formation of secondary dentine are usually much less sensitive. By this formation, as has been seen in the chapter on dentine, many of the tubules are cut off and man}' of the fibrils reach the pulp only by anastomosing with a few in the later formed dentine. The transmission to the nerves of the pulp is thus made more difiicult and imperfect. In all considerations of the sensitiveness of dentine, the purely subjective and hysterical symptoms must be carefully watched for. In many cases slight sensa- tions are so magnified by fear and expectation as to be considered intolerable. In such cases the diversion of attention and the skilful use of suggestions are of more value when coupled "\\'ith delicacy of manipulation and opera- tive skill than any means of obtunding. In such cases, although the operator is positive that the sensations are slight, it ^^^ll never do any good to tell the patient so, or to argue that what is being done cannot hurt. They must be 218 DENTAL PULP made to believe fully that something has been done to destroy the sensitiveness, and then the attention must be concentrated upon something, while the excavation is lightly and skilfully performed. It makes very little difference what is done, but it must attract the attention in order to plant the belief that the sensitiveness has been removed, and then the attention must be diverted until the manipulation is completed. The nerves of the pulp not only respond with sensations of pain from the irritation of the fibrils in the dentinal tubules, but because of their confinement in a calcified chamber and the semifluid nature of the tissue, they are very sensitive to pressure, either increased or decreased. The normal response to changes of temperature, as well as most of the pain in pathologic conditions of the pulp, are probably caused by changes of pressure, through dis- turbance of the blood circulation of the tissue. The nerves of the pulp control the walls of the arteries through the vasomotor reflexes, and also by trophic fibers control the functional activity of the odontoblasts in the formation of the dentine. In a single tooth the irritation resulting from a carious cavity is found to cause the formation of dentine not simply in the region reached by the irritated fibrils, but upon the entire wall of the pulp chamber and apparently also in other teeth. It has seemed possible to the author that in some instances osmotic conditions might be a factor in the pro- duction of pain in the pulp, especially in the early stages of caries. CHAPTER XVI STRUCTURAL CHANGES IX THE PATHOLOGY OF THE PULP Because of its structural peculiarities, as well as the fact that it is a tissue of embryonal character whose function has been chiefl}' performed, the dental pulp is specially sus- ceptible to certain pathologic conditions which produce structural changes. These conditions are hyperemia, inflam- mation, suppuration, and various forms of tissue degener- ation. The dental pulp ofters specially good opportunities for a study of the tissue changes that are characteristic of these conditions, because in the normal state the tissue ele- ments are comparatively widely scattered in an almost struc- tureless intercellular substance. The changes in the blood- vessels, the passage of cellular elements of the blood through the bloodvessel walls and what becomes of them after they enter the tissue can therefore be followed more easily than in tissues which are crowded with cells. HYPEREMIA Hyperemia is defined as an increased amount of blood in a part. It is usually divided into active and passive. In active hyperemia the increase in the amount of blood is due to the enlargement of the arteries supplying the part, or the increase of blood pressure, or both. In passive hyperemia it is due to an obstruction of the veins, so that the blood is not allowed to escape as freely from the part. In case of the dental pulp the conditions which cause an active hyperemia produce, at the same time, a passive one. H^^eremias are also classified as acute and chronic. 220 STRUCTURAL CHANGES IN PATHOLOGY OF PULP Acute Hyperemia. — It is one of the most important of the pathologic conditions of the pulp, because it is one of the most common, and is often the first of a series which result in the final loss of the organ. It is this condition of the pulp which most commonly calls the patient's attention to the presence of a carious cavity. The destruction of the tooth tissue as well as the irritation of the dentinal fibrils by the acid produced, increase the irritability of the cytoplasm of the odontoblasts, and the normal sensory function of the pulp is greatly exaggerated. The response to sudden changes of temperature which constitutes the normal sensory function of the pulp is, in fact, a momentary acute hyperemia, which is immediately recovered from by the return of the normal caliber of the arteries. The reaction is brought about by the vasomotor nerves which control the arteries. As soon as the artery dilates a greatly increased amount of blood is poured into the tissue, and all of the minute capillaries are distended to three or four times their size (Figs. 164 and 165). Because of the semifluid character of the intercellular substance, the pressure is transmitted to the nerves, and a sharp lancinating pain is the result, which lasts until the dis- tention of the bloodvessel subsides, which occurs in a few seconds in normal conditions. When a tooth is exposed continuously to sudden changes of temperature, or to excessive heat, the function becomes greatly exaggerated, and changes which would ordinarily produce no effect will produce acute hyperemia. When the irritability of the fibrils and the odontoblasts has been greatly increased by the action of irritating agents, as in the progress of caries, or when the thickness of the protecting dentine has been greatly reduced, as in abrasion, or when consider- able masses of gold are separated from the pulp only by a thin layer of dentine, the same conditions result. In this stage of hyperemia the only change in the tissue that can be observed under the microscope is the distention of the capillaries and veins, and as soon as the pain has passed the tissue returns to a normal condition. It is apparently, therefore, a functional disturbance, due ACUTE HYPEREMIA 221 to the increased irritability of the cytoplasm of the fibrils, odontoblasts, and probably also of the nerve endings. The rational treatment for such conditions is the removal of the Fig. 164 .'.'l^B m r. T\ I ■''V ' ^m %^' Acute hyperemia. Fig. 165 Acute hyperemia, higher power. 222 STRUCTURAL CHANGES IN PATHOLOGY OF PULP irritation which has caused the irritabiUty, and the complete protection of the tooth from thermal change, until the rest restores the normal function. In order to observe the structural changes, the tooth must be extracted during the paroxysm of pain, and should be cracked and dropped at once into a fixing fluid, allowed to remain there for about twenty-four hours, when the pulp can be removed from the pulp chamber and embedded and sectioned. In this w^ay the injection of the bloodvessels will be preserved, and all of the capillaries and veins w^ill be found crow^ded with corpuscles, and their distention will be proportionate to the severity of pain at the time of the extraction. Acute hyperemia has two possible terminations aside from recovery: (1) If often repeated, it may pass over into chronic hyperemia; (2) if severe enough, it may end in infarction. Chronic Hyperemia. — When the paroxysms of acute hyperemia are often repeated the endothelial cells of the bloodvessel walls lose their power of contractility, and the vessels remain permanently dilated. In this condition the bloodvessels are dilated, even between the paroxysms of pain, and thin w^alls often shoW' pouches and varicosed enlargements. (See Black, American System of Dentistry, vol. i, p. 846.) During the paroxysms of pain, both red and w^hite blood corpuscles are forced through the blood- vessel walls, and areas of breaking down red blood corpus- cles may be found in the tissue. The number of cellular elements in the pulp becomes greatly increased. Whether this increase is due to the multiplication of the connec- tive-tissue cells of the pulp or to the development into tissue elements of w^hite blood corpuscles is a matter upon which opinions differ. In the study of such specimens the author has been unable to escape the feeling that many of the white blood corpuscles develop into tissue elements, w^hich, however, have not the form of the typical connective- tissue cells of the pulp. Fig. 166 is a photograph of a section of such a pulp, and INFARCTION 223 by comparison with Fig. 167 the increase in the number of cellular elements is very apparent. Chronic hyperemias are usually followed after exposure of the pulp by inflammation and suppuration, but if this Fig. 166 Chronic hyperemia, showing increase of cellular elements is prevented by the treatment of the cavity, the tissue is likely to undergo fibrous or other degeneration. Infarction. — Complete infarction of the dental pulp is rare, but partial conditions are not uncommon. The con- dition is comparable to the conditions that occur in the brain 224 STRUCTURAL CHANGES IN PATHOLOGY OF PULP and other places supplied by end arteries, without anasto- mosis, when the vessels carrying the blood from the part are completely occluded. The following clinical picture will occasionally be encoun- tered with such a history. A tooth beginning to ache suddenly, perhaps, because of a sudden exposure to change of temperature, continues to ache violently several hours, the pain being described as acute and lancinating, and so severe as to be almost intolerable. Nothing done relieves the symptoms in the least. Finally, the pain stopped almost as suddenly as it began. The next morning the tooth is more or less red in color, and by the time it reaches the operator it begins to turn dark. What has happened is represented in the following tissue changes. An extremely acute hyperemia has occurred, the dilatation of the arteries entering the apical foramina have compressed the more delicate walls of the veins so as to occlude them completely, and greatly increased blood pressure has distended all of the capillaries and veins, forcing the red and white blood corpuscles, as well as the serum, through their walls, filling the tissue. Complete stasis has resulted after a few hours in the death of all the tissue elements, at which time the pain stopped. The serum has dissolved the hemoglobin from the red blood corpuscles, filtered through the dentinal tubules, discoloring the dentine and showing through the enamel. Small areas of partial infarction are found in many specimens after severe paroxysms of acute hyperemia^ which may be recovered from entirely. The severe pain which occasionally results from the application of arsenic for the devitalization of pulps is due to the acute hyperemia which is induced. The removal of the arsenic application will not alleviate the pain, which can be subdued only by the immediate extirpation of the pulp. Inflammation. — Inflammation of the pulp occurs ordin- arily only after exposure, and follows a chronic suppurating course, progressing along the veins; the line of demarcation between the normal and inflammatory areas often being PULP NODULES 225 quite sharply marked. In the first stages the white cor- puscles are seen along the walls of the vessels, passing through the walls into the tissue in increasing numbers until the tissue becomes a solid mass of cells and serum breaking down into pus. This progresses until the entire tissue is destroyed. There is the greatest difference in the rapidity with which the stages follow each other and the extent to which the inflammation spreads through the tissues before the breaking down begins. This is probably due both to the character of the invading microorganisms and the resistance of the individual. These conditions are illus- trated in Figs. 167, 168, 169, 170, 171, and 172. The for- mation of pulp nodules is often noted in the deeper part of the tissue in which inflammation is progressing (Fig. 169). Occasionally centres of inflammation, progressing to abscess formation, are found within the substance of the pulp (Fig. 174). These are apparently true intrapulpal abscesses and present the characteristics of miliary abscesses in any other tissue. Degeneration. — The embryonal character of the pulp tissue renders it specially susceptible to degenerative changes, but the degenerative changes of the dental pulp have never been adequately studied. It is extremely difficult to obtain material. Teeth without histories are practically useless, and large numbers of specimens are necessary. Pulp Nodules. — In the cutting of large nuqabers of pulps for the preparation of class work the author has been im- pressed by the frequency with which hard nodules occur in the tissue. These are apparently of several varieties, some of which are calcified and others are not. They usually occur in the coronal portion of the pulp near the opening of the canals. They often occur in specimens in which the tissue is otherwise normal. Fig. 176 shows a section with a small, almost spherical nodule in the centre of the lower part of the coronal portion. For a number of sections the nodule was cut through as if it had a soft periphery, then it took a nick out of the razor and was pulled out of the tissue, the subsequent section showing the hole it had left (Fig. 177). 15 226 STRUCTURAL CHANGES IN PATHOLOGY OF PULP Fig. 167 Beginning inflanunation. ^^"^ - - ^— ^-->-o<^/ 'W ^. ^^$ t§M 4 Yf l^^ jtaWia^^ Cells from the growing tip of a chestnut seedling. with the growth of the cell until there is a rim of cytoplasm in contact with the cell wall, and a central mass of cytoplasm surrounding the nucleus and connected with that at the periphery by fine threads. In still further growth these threads are broken, the nucleus is pushed to one side, and the whole central portion becomes one huge vacuole. There is now a cell wall, with a layer of cytoplasm covering its inner surface, which is kept in reaction with the nucleus by streaming around and around. This flowing of the cytoplasm in plant cells may be easily observed in the dehcate stamen 240 INTERCELLULAR SUBSTANCES hairs of the ordinary Spiderwort, or in the cells of the water plants Chara or Nitella, which are easily found in most ponds. In this example it is seen that the cytoplasm remains in contact with the formed material which it produces for support, and that it is only sufficient in amount to form and maintain this material. In general histology it has already been noted that the cells of connective tissue are very similar, and that the tissues differ chiefly in the character and arrangement of the intercellular substances. It has also been emphasized that the connective tissues all originate from a common form of embryonal connective tissue, or mesenchyme, and change from one form to another in development. These mutations of the connective tissues are its most striking characteristic, and must be clearly grasped if the bone, as an organ of support, is to be understood. For instance," embryonal connective tissue is transformed into fibrous tissue; fibrous tissue becomes arranged in a definite mem- brane, and is transformed into cartilage, which is again removed and transformed into bone. All these changes take place to meet the requirement of mechanical conditions and influences. If the subcutaneous tissue of an embryo be examined in sec- tions (Figs. 184 to 199) the cells will be found to be irregular masses of cytoplasm with a nucleus in the central portion, and fine projections stretching out in all directions through an almost structureless intercellular substance. The fine pro- jections of the cytoplasm meet those of the adjoining cells and form a network holding everything together. Because of the nature of cytoplasm, however, these possess very little strength, and very soon fine thread-like fibers are found appearing in the intercellular substance in contact with cells. These unite with each other, forming continuous fibers, and very soon a strong netw^ork is produced which is entirely dependent upon the cytoplasm of the cell which has formed and maintains it. If this tissue is now subjected to pressure and strain, the cells become flattened out and squeezed between the bundles of fibers, which take on INTERCELLULAR SUBSTANCES 241 parallel directions, and so a tendon is formed. A tendon must be considered as a highly specialized form of connective Fig. 184 Fig. 185 --KK<', ^^:^- -^ ^^i-T-S Embn,'onal connective tissue in an early The same, a little more developed, stage of development, showing the cellular showing the cellular elements length- elements embedded in the ground sub- ening in a common direction, stance. Fig. 186 ^--^"-ir^^^^--- - The cells developed in spindle forms, fibroblasts with long filaments extending from either end. Fig. 187 The developed white fibrous tissue. tissue, arranged to supply tensile strength. The degree of specialization of the tissue is judged by the extent to which 16 242 INTERCELLULAR SUBSTANCES its characteristic features are developed, either in quantity or quality. In the tendon the fine strong fibers have l)een Fig. 188 ^ Older white fibrous tissue, in which the cells are no longer seen, and showing the wave like course of the fibers. Fig 189 Coarse white fibers, made up of bundles of the fine fibers, and showing the mode of division by splitting off of a portion of the fibers of the bundle. Fig. 190 Coarse fiber breaking up into fine fibers. gathered into bundles; a round nucleus would occupy too much space. It has, therefore, become elongated and more INTERCELLULAR SUBSTANCES 243 or less rod-shaped, and the cytoplasm has been squeezed out into thin leaf-like projections between the bundles. Fig. 191 Cross-sections of coarse fibers, showing some of their various forms. Fig. 192 Fig. 193 & Reticular or elastic fibers, showing the mode of division and the multipolar, or irregular, star forms of the cells at the divisions. Fig. 194 ^•/ ^ o Cross-sections of the reticular fibers, showing some of their forms. Connective-tissue cells from which reticular fibers are developed. Each cell is in contact with several fibers, and each fiber in contact with the cytoplasm of cells which have produced them. 244 INTERCELLULAR SUBSTANCES Fig 195 Fig. 196 Network of elastic fibers from the point Network of elastic fibers teased out of reflection of the mucous membrane of from elastic tendon, and showing the the lip from the gums. usual mode of division. Fig. 197 / ^s-^ ^yf^S' Elastic fibers, showing their disposition to curl up when cut or broken. INTERCELLULAR SUBSTANCES 245 It must be supposed that there is a constant reaction between the substance of the formed material and materials produced by the metabolism of the c^i:oplasm. In patho- logic conditions the metabolism of the cytoplasm is dis- turbed, and there is a consequent change in the quality of the fibers. So in some pathologic conditions a relaxation and loss of tone is found in tendons and ligaments. In inflammations of the gingivae the fibers become relaxed and stretched, so that the gingivse are everted, but return to their normal condition when the pathologic condition has subsided, and the cells regain their normal metabolism. Fig 198 Fig. 199 ■:-:>^'i-~/ir<'' . - r. ,, '^ ' ■',■■ fc?:.-; .- :: .\.:::-:^- :-, --^^ '.:'.::-.. r-^.---..f^^^-& Cross-sections of elastic fibers, showing their forms as seen in Tissue of the dental pulp, in which the de- a group passing between coarse velopment of the cells is not followed by any white fibers. considerable formation of fibers. To sum up w^hat has been said, it is apparent that both phylogenetically and ontogenetically, intercellular substances have been produced and are maintained by cells in response to mechanical influences and to meet mechanical conditions. In all higher animals certain tissues, the connective tissues, have been set apart for this purpose, and the cells have been specialized to respond to mechanical stimuli and develop an intercellular substance adapted to the condition. This makes the supposition necessary that an embryonal connec- tive-tissue cell may develop into any specialized form and that the kind of cell into which it develops will be determined by the character of mechanical stimuli which it receives. Just as the epithelial cells have been specialized to respond to the environments of light stimuli, vibration of the air, pressure, and chemical action which connect the organism with its environment, connective-tissue cells have been 246 INTERCELLULAR SUBSTANCES specialized to respond to mechanical stimuli, by the pro- duction of formed materials adapted to the mechanical conditions. These conceptions are fundamental to an understanding of bone structure and growth, and the muta- tions of connective tissue in general. In no branch of histology is a clear conception of inter- iellular substances and the relation of cells to them as mportant as in the study of the teeth and their associated structures. Caries cannot be understood unless these fundamental ideas have been appreciated, and many state- ments in dental literature would never have appeared if the nature of intercellular substance and the relation of cytoplasm to it had been understood. CHAPTER XVIII BONE Defimtion. — Bone may be defined as a connective tissue whose intercellular substance is calcified and arranged in layers around nutrient canals or spaces. The cells are placed in cavities, lacunae, between the layers, and receive their nourishment through very minute channels, canal- iculi, which radiate from them and penetrate the layers. STRUCTURAL ELEMENTS The structural elements of bone are : 1. Bone matrix, or intercellular substance, which is always arranged in layers or lamellte. 2. The bone cells or bone corpuscles which are embedded in the matrix between its layers. 3. Lacunae, or the spaces in which the cells are found. 4. CanalicuH, or the channels through the matrix by which the embedded cells receive nourishment. Bone Matrix. — The bone matrix is composed of a dense organic basis of ultimately fibrous character which yields gelatin upon boiling with water. With this inorganic salts are combined in a weak chemical union, forming the hard substance of bone. By treatment with acids the inorganic salts can be removed, leaving the organic basis which retains the form of the tissue. In this condition the rigidity of the bone is destroyed. On the other hand, by calcining at red heat the organic basis can be removed, leaving the inorganic substances which retain the form of the tissue. In forma- tion the organic basis is apparently formed first, and then the salts of lime are combined with it, through the agency of the formative cells, or osteoblasts. 248 BONE Bone Corpuscles. — Bone corpuscles are the cells lying in the laciuiie. Each cell contains a single well-defined nucleus, lying in the centre of a granular cytoplasm. The cell appar- ently completely occupies the lacunae, and from the central mass fine projections of cytoplasm extend through the Fig. 200 From a section through the bone of a roebuck. The lacunae are seen from above, and are filled with coloring matter. In places small dots are visible, which represent the cross-sections of bone canaliculi. (850 X) (Szymonowicz.) canaliculi, which brings the bone corpuscles in intimate relation with certain area of bone matrix. The processes of one cell anastomose with those of its neighbors through the canaliculi, so that there is a continuous network of living cytoplasm throughout the matrix. CANALICULI 249 Lacunae. — The lacunae are flat oval spaces about 20 microns long, 10 microns wide, and 5 or 6 microns thick. Their shape, therefore, in sections depends upon the way in which they are cut as illustrated in Figs. 200 and 201. When Fig. 201 From a section through the bone of a roebuck. The lacume are seen from the side. (850 X) (Szymonowicz.) cut lengthwise they would appear as about 20 microns long and 6 wide in profile, or as about 20 microns long and 10 wide when seen from above. Canaliculi. — These radiate from the lacunae in all direc- tions, opening into them by larger channels which branch 250 BONE and divide, becoming smaller as they pass farther into the matrix. They anastomose freely with those from adjoining lacunae. THE VARIETIES OF BONE There are three varieties of bone differing in the arrange- ment of these structural elements. These are subperiosteal, Haversian system, and cancellous bone. Subperiosteal Bone. — This form of bone must be regarded as primarily a formative arrangement and more or less transitory, in which the layers are arranged parallel with the surface, and under a formative membrane. It contains canals (Volkmann's canals) with bloodvessels (Fig. 202), con- nective tissue, etc. These penetrate the layers which are never arranged concentrically around them. It is always thin, that is, composed of comparatively few layers, and when a considerable thickness is formed it is cut out from within by absorptions beginning in the canals, and bone is rebuilt with layers arranged concentrically around the chan- nels formed. In this way subperiosteal bone is converted into the second form. Haversian System Bone.^In this variety the lamellae are arranged concentrically around canals which contain blood- vessels, nerves, and embryonal connective tissue, and from which the cells in the lacunae are nourished (Fig. 203). These canals are, in general, parallel with the surface or the long axis of the bone and anastomose with each other. A canal with the layers arranged around it constitute an Haversian system. Between the Haversian systems are remains of the subperiosteal layers (insterstitial lamellae) that were left by the absorption, and for that reason have been called fundamental lamellae. They have also been called ground lamellae. Haversian system bone is often called compact bone, and makes up the greater part of the shafts of the long bone, and the plates of the flat ones. It is never allowed to become greater in thickness than is necessary for strength, and when sufficient thickness has been formed, the deeper CANCELLOUS BONE 251 part is cut out by absorptions in the Haversian canals, con- verting them into large irregular spaces. The formation of a few layers around these spaces transforms the second type into the third or cancellous bone. Ftg. 202 Fig. 203 Subperiosteal bone, showing Volkmann's canals. Haversian system bone; a, Haversian canals. Cancellous Bone. — In this variety the lamellae are arranged in delicate plates surrounding large, irregular nutrient or marrow spaces. These are filled by embryonal connective tissue and contain bloodvessels and nerves. The plates of cancellous bone are not arranged at haphazard, as might be supposed from a casual observation of sections, but are disposed in definite arrangement, which is determined by the directions of stress on the compact bone which they 252 BONE support. (See illustrations in Chapter XVIII.) They are not permanent and unchanging, but are continually being rebuilt in new directions, in response to the mechanical con- ditions to which the bone as a supporting organ is subjected. THE ARRANGEMENT OF BONE Compact Bone. — A knowledge of the structural elements of bone can best be obtained by the study of sections ground from the shaft of a long bone. An old dry bone should be sawed across, near the middle of the shaft, in two places, so as to cut out a ring about a quarter of an inch thick. Then saw the ring through in two places with an arc of about a quarter of an inch on the outer surface. From this two slices should be sawed out, one transverse to the long axis of the bone, the other parallel with it. These are ground to not more than 8 or 10 microns in thickness and mounted in hard balsam. From a study of these two the arrangement of the lamellae, and the shape and character of the lacunse can be made out. Upon the outer surface of the transverse section will be found a larger or a smaller number of layers of subperiosteal bone which encircle the shaft, and consequently are called the circum- ferential lamellae. The number of these layers will depend upon the position from which the section is taken, and the age of the bone. If the bone is increasing in circumference at the point from which the section is cut, there will be a considerable number of layers, and they will be easily seen. If the bone has been growing smaller in circumference at the point, there will be very little of subperiosteal bone, and it will be comparatively hard to recognize. The greatest part of the section will be made up of Haversian systems, in which from two to three to five or six layers are arranged around an Haversian canal. The lacunae appear as irregularly oval spaces about 5 or 6 microns across and 15 to 20 microns in length. From them a great many minute canals radiate through the matrix both toward the Haversian canal and PLATE X K -^ f: ^ if. V- X W ■ "- ^ V "^ "^ *^» - ' ^ * 1 \ <^ M -4-rf ^ X - i i s ^ ^ , f ^ < V .' .A. < Froni a Ground Cross-section of the Diaphysis of the Human Metatarsus. (Szynionowicz. ) a, outer ground lamellae; ;>, inner ground lan:iellae; c, Haversian laniellae; d, interstitial laniellae. All canals and bone cavities are filled with coloring nnatter and appear black. (90 X) COMPACT BONE 253 away from it. The character of these canaliculi can only be appreciated by seeing them. They are filled in life by pro- jections of the protoplasm of the bone corpuscles. They are suggestive of the rootlets of plants running through soil, and as in that case the rootlets are absorbing material from the soil and reacting with it, in this case the protoplasmic con- tents of the canaliculi is reacting with the matrix, maintain- ing its quality. The portion of matrix through which the canaliculi from one lacunae extend belongs to the bone corpuscles which occupies the lacunse, as will be seen later. These cells have been enclosed in the matrix which they have formed. Between the Haversian systems will be found a few layers of interstitial or fundamental lamellae. They are the remains of layers which were formed under the periosteum and were not entirely destroyed when it was replaced by Haversian systems (Plate X). The amount of interstitial lamellae varies greatly in different specimens, as will be seen by comparing figures. The Haversian canals anastomose with each other; this will be seen in many specimens. Many Haversian sj^stems will be found imperfect in form, as, for instance, those shown in Plate X. This means that after these systems were com- pleted, absorptions occurred in a neighboring canal which attacked the layers of the system, and later a new system was formed in this space by the deposit of concentric lamellae. While bone is thought of as a hard and fixed tissue, it is con- tinually being built and rebuilt in this way. It is onty by the understanding of these possibilities that we get the ideas that bone, while hard and rigid, is a plastic tissue and is con- tinually being moulded by mechanical conditions to which it is subjected. It will be seen also that the arrangement of the lamellae becomes a record of the changes that have occurred in the formation of the tissue. The inner boundary of the section next to the marrow cavity will show a few layers parallel with the surface. These are known as the inner circum- ferential lamellae. It is a mistake, however, to think of them as surrounding the marrow cavity in the same sense as 254 BONE the outer circumferential lamellse surround the bone. If the section has been cut at a little distance from the centre of the shaft, it will have been noted that the marrow cavity is penetrated by very delicate spicules, and that in fact the marrow cavity is produced by the spaces of cancellous bone, becoming larger and larger until they become one continuous space. The inner circumferential lamellae are therefore the layers which have been formed around an enlarged nutrient or marrow space. Cancellous Bone. — The cancellous bone can best be studied in decalcified sections. A field from the central portion of a flat bone will show its typical arrangement. It is made up of delicate flattened spicules surrounding larger or smaller irregular spaces which connect with each other very freely. Each spicule is composed of a few lamellae which are arranged around the space. The structure of the spicules often becomes complicated by absorptions and rebuildings which have occurred to change their direction. The tissue which fills the spaces is a delicate, embrj^onal connective tissue in which osteoblasts and osteoclasts appear in response to mechanical conditions. It is richly supplied with blood- vessels, nerves, and lymphatics. The lacunae and canal iculi are in no respect different from those of the Haversian system and subperiosteal bone. CHAPTER XIX BONE FORMATION AND GROWTH Bone is one of the latest tissues to be formed, and is always developed from an antecedent connective tissue of less specialized character. According to the character of the antecedent tissue, bone formation is of two varieties — the formation from cartilage, or endochondral bone forma- tion, and that from fibrous connective tissue, without the intervention of cartilage, or endomemhranoiis bone formation. Endochondrial Bone Formation. — ^All of the bones of the endo- skeleton are preformed in cartilage. The transformation of cartilage into bone is rather a substitution than a transfor- mation, for the original tissue is destroyed in the process, aiKl a new and more highly specialized one substituted for it. Before ossification begins the cartilage has taken on the general form of the bone and is covered by a definite peri- chondrium. Ossification begins at separate centres and progresses through the cartilage, but the separate centres do not unite until the bone is about fully formed. In the long bone there are usually three centres — one near the centre of the shaft, forming the hypophysis, and one near either end, forming the epiphysis. These remain separated by a layer of cartilage until the length of the bone has been fully formed. The first indication of the transformation of cartilage into bone is an increase in the size of the lacunae and in the amount of cartilage matrix, which also shows changes in character, having lime salts deposited in it. The car- tilage c^lls enlarge and show signs of degeneration, the lacunse become arranged in rows, and as they increase in size, more in the direction parallel with the axis of the cartilage, the amount of matrix separating them is reduced. 256 BONE FORMATION AND GROWTH By this time the perichondrium, on the surface of the car- tilage opposite to the centre, has developed osteoblasts which begin the formation of subperiosteal lamellse upon the sur- FiG. 204 Hyaline cartilage Area of calcification ).',.} Osteogeneiic tissue Perichondral "*""" bone Capsules containing •many cartilage cells From a longitudinal section of a finger of a three-and-a-half-months human embryo. Two-thirds of the second phalanx is represented. At X a periosteal bud is to be seen. (85 X) (Szymonowicz ) face of the cartilage, and the perichondrium is transformed into periosteum. Opposite the centre osteoclasts appear, cutting into the cartilage, followed by buds of embryonal tissue. The osteoclasts dissolve away the remains of the EXDOCHOXDRIAL BOXE FORMATION 257 cartilage matrix, opening up the spaces between the lacunae and converting the rows of lacunae into irregular channels or primary marrow spaces. Upon the spicules of calcified cartilage matrix, osteoblasts arrange themselves and begin to lay down lamelhie of bone. These changes progress from Fig. 205 Cartilage cell The place marked X in the preceding figure with stronger magnification. (185 X) (Szymonowicz). the centre in both directions, and all stages, from the typical hyaline cartilage to the formation of bone, may be seen in one section. These stages are illustrated by Figs. 204, 205, and 206. From now on the bone grows by progressive transforma- tion of cartilage and by the growth of bone under the peri- osteum, which will be considered under bone groT\i:h. 17 258 BONE FORMATION AND GROWTH Endomembranous Bone Formation. — The bones which are not preformed in cartilage are formed directly from fibrous Fig. 206 Periostenm^^^] Periosteal bntl Blood-i'essris filled witJi^: blood^ Calcified |/'/^ij'r* ' /JJ cartilage'*^' '*' " Enlarged cartilage cells From longitudinal section of a finger of a four months embryo. Only the diaphysis of the second phalanx is represented. (85 X) (Szymonowicz.) tissue. This is well illustrated in the mandible. In the region of Meckel's cartilage and between it and the develop- ing tooth germs the mesenchyme begins to show signs of ENDOMEMBRANOUS BONE FORMATION 259 specialization. Delicate fibers appear in the intercellular substance. Along these the connective-tissue cells arrange themselves, and, taking on the form of osteoblasts, begin to Fig. 207 i3 C <9\0, ^'.^>'^, N. ^ ^'^-v.- -^ -^( _Primar\' marrow space. Osseous tissue. Section through the lower jaw of an embr>-o sheep (decalcified with picric acid). At a and immediately below are seen the fibers of a primitive marrow cavity Ij^ing close together and engaged in the formation of the ground substance of the bone, while the cells of the marrow cavity, with their processes, arrange themselves on either side of the newly formed lamella and functionate as osteoblasts. (Bohm, Davidoff, Huber.) (300 X) lay down bone lamellae (Fig. 207). These stretch out through the mesenchyme, forming a network of delicate spicules, until they surround Meckel's cartilage, and grow up to the 260 BONE FORMATION AND GROWTH buccal and the lingual of the tooth germ. As soon as this network of bone lamellee containing embryonal connective tissue, in its primary marrow spaces, begins to take on definite form, there is a specialization of the mesenchyme surrounding it, developing into fibrous tissue which becomes a periosteum. From this time onward the formation of bone progresses, as will be described under the growth of bone. Bone Growth. — If sections are cut transversely through the shaft of a long bone from a fetus, the surface will be found to be covered by a well-formed periosteum, which is actively laying down layers of subperiosteal bone. The central portion of the bone is made up of a network of spicules surrounding primary marrow spaces, there being no true marrow cavity. The formation of the subperiosteal laj^ers does not progress at a uniform rate at all points* on the circumference, but they are piled up at certain points forming longitudinal ridges with grooves between them. These grooves become arched across, enclosing part of the connective tissue of the inner layer of the periosteum, and contain bloodvessels and nerves. Soon after these spaces are enclosed absorptions begin in their walls, destroying a large part of the subperiosteal lamellae and forming primary marrow spaces. As soon as these spaces have reached a certain size the absorptions stop, and osteoblasts appear upon the wall of the space and begin to lay down lamellae upon its circumference, until an Haversian system has been produced with an Haversian canal at its centre. In this way the bone increases in diameter, and this process continues until a considerable thickness of Haversian system bone is formed. In all bone growth there is the alternation of formation, destruction, and rebuilding, and it must be remembered that this continues as long as the bone functions as an organ of support. As the shaft becomes larger the primary marrow spaces at the centre are enlarged by the absorption, and a few lamellae are laid down again upon their walls, until finally in the central portion of the shaft the true marrow cavity is formed. As -the thickness of Haversian system bone becomes greater, absorptions occur GROWTH OF MEMBRANE BONES 261 in the Haversian canals, cutting out large, irregular channels, around which a few lamellse are laid down, and so the Haver- sian system bone becomes converted into cancellous bone and is opened into the marrow cavity as it grows larger. Growth of Membrane Bones. — The growth of the membrane bone progresses in a very similar way. As soon as the periosteum is formed subperiosteal bone is laid down and converted into Haversian system bone, forming the compact plate of the surface, leaving the cancellous portion first formed at the centre. When a certain thickness of compact bone has been formed, absorptions occur in the Haversian canals, converting the deeper portions into cancellous bone. This process may be reversed. Absorptions may occur under the periosteum, cutting deeply into the Haversian system bone, and then a few subperiosteal layers be laid down upon it. When this occurs lamellae are laid down around the marrow spaces, converting the cancellous bone into Haversian system bone to maintain the required strength. In this way the bones are moulded into shape, adapting them to the mechanical conditions to which they are sub- jected. There is an oscillation between formation and destruction, by which the balance adapted to the mechanical conditions is maintained. It has often been noted that bones are never allowed to become more bulky than is necessary to perform their function. CHAPTER XX PERIOSTEUM^ Definition. — The periosteum is the formative and pro- tective membrane which covers the outer surface of the bone. All periosteum has certain structural characteristics in common, but because of structural differences two classes are recognized — attached and unattached — each of w^hich may be simple or complex. Periosteum may thus be classi- fied as follows: 1. Unattached simple. 2. Unattached complex. 3. iVttached simple. 4. Attached complex. Function of Periosteum. — The importance to the dentist of a knowledge of the structure and function of the periosteum can scarcely be exaggerated. It has been the knowledge of this tissue and its function that has led to all the advance- ment in bone surgery of modern time. Repair and regenera- tion of bone is largely accomplished through its agency. The periosteum forms the immediate covering of all the bones and is continuous over their entire surface except the portion covered by cartilage. Each bone, therefore, has a periosteum of its own which does not continue around the articulation to the bones with which it joins. Bones that are united by suture are, however, covered by a common periosteum. If the flesh and overlying tissues are carefully 1 In the presentation of this chapter it is impossible adequately to express my indebtedness to Dr. G. V. Black. Almost all of the illustrations are taken from The Periosteum and Peridental Membrane, published by him in 1887. I have always felt that this book had never received the attention it deserves. Only one thousand copies of it were printed, and they were not sold until the orthodontists exhausted the edition. The book is now entirely out of print and is very difficult to obtain. I have studied this book for years and have repeated almost all of the work described in it, but I have felt that it was impossible for text-book purposes to improve upon the illustrations. FUNCTION OF PERIOSTEUM 263 removed from a long bone, the periosteum will be seen as. a smooth white, lustrous membrane, having much the appearance of a tendon on most of its surface. But at some places which correspond to the positions where muscles or fascia were attached it appears ragged and dull, for the tissues had to be cut to separate them from the outer layer of the periosteum, to which they were firmly adherent. In all other places the tissues separate easily in dissection; in fact, are not attached at ah, except by the lightest of areolar tissue, which is very easily broken, and the tissues may be separated from the surface of the membrane with the finger or the handle of a scalpel. Now, if the periosteum is slit along a smooth surface with the scalpel and the handle inserted between the bone and the membrane, it will be found to separate readily from the bone over most of its surface. If the process is watched closely, little strings will be seen apparently running from the periosteum to the bone, and being broken as they are separated. These are mostly small bloodvessels which are running into canals in the bone. In this process the periosteum seems like a closely adapted sac or elastic glove, clothing the surface of the bone, as if surrounding it in a fibrous bag. If the separation of the periosteum from the bone is continued, it will be found that it does not separate as easily in all places. As the articular ends are approached it becomes suddenly fastened to the underlying bone, and the blade of the knife must be used. The periosteum now appears as a very thin, tough, and inelastic membrane, that is torn with difficulty, but it is so thin that it is difficult now to separate it from the bone without cutting it through. When this point of attachment is reached it seems that the periosteum is sinking into the substance of the bone, and from the examination of its structure it is found that this is practically what has happened. Comparing the periosteum to a sac surrounding the bone, it is found sewed firmly down at the margin of the cartilage around the articular ends. Besides the attach- ment around the cartilage, the periosteum will be found 264 PERIOSTEUM adherent in the following positions: Where muscles or fascia are attached to the outer layer of the periosteum; where it approaches the insertion of tendons or ligaments; and where the skin or mucous membrane seem attached to the underlying bone, as around the auditory meatus, the gums, mucous membrane of the nose, etc. In all such positions the periosteum is firmly attached to the bone — in fact, becomes a part of it — and through this medium the connections between muscles, fascia, etc., and the framework of the skeleton is accomplished. This feature of the anatomy of the periosteum has never been studied in the detail it deserves, especially by the dentist. It is of the greatest importance in the manage- ment of the diseases of bone, especially those involving the formation of pus, for these lines of attachment deter- mine the direction in which the pus will proceed along the surface of the bone. When pus generated within the bone reaches the surface, it will lift an unattached periosteum and run along the surface until it reaches a line of attach- ment. Here it can penetrate the periosteum more easily than it can separate it from the bone. W^hen a line of attach- ment is reached, therefore, the direction of the burrowing is de- termined by the attached areas. The pus penetrates the peri- osteum more easily than it separates its attachments from the bone, but it lifts the unattached periosteum so easily that it will often run along a line of attachment for a long distance. These factors often become of great importance in deter- mining the position in which alveolar abscesses will point. For instance, if an abscess from a bicuspid root, or the mesial root of a molar, reaches the surface of the bone above the attachment of the buccinator, it cannot pene- trate its attachment and pass downward to open on the gum, but may run out over the surface of the muscle and open on the cheek, producing the crow's foot scar so often seen. An abscess from an upper cuspid may reach the surface of the bone in the canine fossa between the attach- ments of the nasalis and canius, and lift the periosteum extending upward, and open at the inner canthus of the eye SIMPLE UNATTACHED PERIOSTEUM 265 between the orbicularis and the angular head of the quadratus labii superioris. If these abscesses had been reached with a lance, through the mucous membrane, at the proper time, a disfiguring scar would have been avoided. Accurate knowl- edge of the attached layers of the periosteum would have made it certain that they could never point in the mouth cavity without assistance. Layers of the Periosteum.— Periosteum is always composed of two distinct layers: 1. An outer or fibrous layer, which is essentially protective and to which muscles and fascite are attached. This may be either simple or complex. 2. An inner or osteogenetic layer which is essentially the vital functioning layer, and is, as its name indicates, con- cerned with the formation of bone. This may be either simple or complex. The Structural Elements. — The periosteum is composed of the following structural elements: 1. White fibers in coarse bundles (in the outer layer). 2. White fibers in very fine bundles (in the inner layer). 3. Elastic fibers. 4. The penetrating fibers, or white fibers of the periosteum, that in the growth of bone are included in its substance. 5. Embryonal connective-tissue cells. 6. Osteoblasts or bone forming cells. 7. Osteoclasts or bone absorbing cells. Unattached Periosteum. — In the unattached periosteum the inner layer is always simple, and the outer layer may be either simple or complex, depending apparently upon the requirements of protection. In general, the more exposed the position the thicker is the layer, and the larger and stronger the bundles of fibers of which it is composed. Simple Unattached Periosteum. — Where the periosteum is covered by a thick layer of muscles which are not attached to it, as in the thigh, the thinnest and simplest form of periosteum is found. An illustration, drawn by Dr. Black, of the periosteum from the femur of a kitten will illustrate its structure (Fig. 208). The outer layer is composed chiefly of 266 PERIOSTEUM bundles of white fibers, most of which run in a direction parallel with the long axis of the bone. The bundles are com- paratively small and much flattened, so as to be quite rib- bon-like. The inner layer contains a much greater number of cells lying among extremely delicate fibers. In its outer portion many of the cells are embryonal in character. In contact with the surface of the bone is a continuous layer of xl Fig. 208 :K Non-attached periosteum from the shaft of the femur of the kitten: B, bone; 0, layer of osteoblasts. In the central portion of the figure they have been pulled slightly away from the bone, displa>-ing the processes to advantage. It will be observed that the fibers of the periosteum do not enter the bone, a, inner layer of fine white fibrous tissue (osteogenetic layer) showing the nuclei of the fibroblasts and a number of developing connective-tissue cells, which probably become osteoblasts; c, outer layer, or coarse fibrous layer, in which fusiform fibroblasts are also rendered apparent by double staining with hematoxylin and carmine; d, some remains of the reticular tissue connecting the superimposed tissue -with the periosteum. (y_r immersion.) (Black.) osteoblasts which are building subperiosteal bone in the young animal, processes of their cytoplasm extending into the canaliculi of the matrix which they have formed. At one point in the illustration the osteoblasts are pulled off from the surface of the bone and show these processes stretched out of the canaliculi. ATTACHED PERIOSTEUM 267 Complex Unattached Periosteum. — In some places, espe- cially where muscles or tendons perform sliding movements over an unattached periosteum, the outer layer, instead of being simple, may be very complex. This is illustrated in Dr. Black's drawing (Fig. 209), from the periosteum of the tibia of a young pig. In this instance the outer layer is ^: Fig. 209 Periosteum from the shaft of the tibia of the pig. lengthwise section, showing the complex arrangement of fibers in the coarse or outer fibrous layer that sometimes occurs under muscles that perform sliding movements upon it: B, bone; O, layer of osteoblasts. The tissue has been pulled shghtly away from the bone in mounting the section, and part of the osteoblasts have clung to the bone, some have clung to the tissues, while others are suspended midway, their processes clinging to each, a layer of fine fibers; inner or osteogenetic layer of the periosteum; 6, first lamina of the coarse or outer fibrous layer, the fibers of which are, in this case, circumferential, exposing the cut ends. It will be observed that there are ten lamina in the make up of the outer layer, the lengthwise and circumferential fibers alternating. The ones marked / and i are very delicate ribbon-like forms, which have shifted from their normal posi- tion in the mounting of the section, so as to present their sides to view instead of their ends, thus displaj-ing their structure to advantage. The illustration shows how readily separable these lamina are. I, reticular tissue. (jV immersion.) (Black.) composed of very much flattened bundles of white fibers, arranged alternately longitudinally and circularly. Ten layers may be counted in the section. The inner layer is of the same character as in a simple specimen. Attached Periosteum. — The attached periosteum differs from the unattached by having the fibers of the inner layer 268 PERIOSTEUM arrano:ed in bundles, around which the bone matrix is deposited hy the osteobhists, embedding them in the sub- stance of the matrix and calcifying them with it. These fibers constitute the penetrating fibers. They were first described by Sharpey, and have been called Sharpey's fibers. He, however, apparently did not understand their importance or manner of formation. The fibers of the inner layer are built into the substance of the bone in this way wherever tissues are attached to the outer layer of the periosteum. ^ '^■^ - ' N ,*• Simple attached periosteum: a, bone; h, osteoblasts; c, fibers of the inner layer; D, bloodvessels of the inner layer; E, outer layer; F, muscle fibers attached to outer layer. (Black.) Simple Attached Periosteum.— Where the pull of tissues attached to the outer layer of the periosteum is in one direction, the fibers of the inner layer are inclined in the same direction (Figs. 210 and 211). As the surface of the bone Fig. 211 A ptiotomicrograph of an attached periosteum similar to Fig. 210. From the alveolai process of a sheep. (About 80 X) Fig. 212 r' a Attached periosteum from beneath the attachment of the muscles of the lower lip of the sheep: .4., bone; S, osteoblasts, with the fibers emerging from the bone between them; C, inner layer with fibers decussating and joining the inner side of the coarse fibrous layer in opposite directions (this is rather an unusual form of this layer of the periosteum); Z), coarse, fibrous layer; E, attachment of muscular fibers. (Black.) 270 PERIOSTEUM is approached the fibers are gathered into strong bundles to be inserted in the bone, the osteoblasts covering the surface of the bone everywhere between the fibers. The outer and inner layers are united by the interlacing of their fibers. At the junction of the outer and the inner layers many bloodvessels are seen. Complex Attached Periosteum. — Where the pull upon the outer layer is in many directions, the fibers of the inner layer, after emerging from the bone, break up into smaller bundles and anastomose in all directions, arching around to interlace with the fibers of the outer layer, and in this way they sustain force in all directions (Fig. 212). This is illustrated in Dr. Black's drawing of a section of attached periosteum from beneath the attachment of the muscles of the lower lip of a sheep. CHAPTER XXI THE ATTACHMENT OF THE TEETH That the teeth are not a part of the osseous system, but are appendages of the skin, supported in man by a special development of bone forming the alveolar ridges of the maxillary bones, is as well established as any fact concerning human dentition. The work of Oscar Hertwig, published in 1874, established very clearly the homology existing be- tween the teeth and the dermal or placoid scales of the ganoid, silurioid, and dipnoan fishes, both as to similarity of structure and development. Much has been written descriptive of the teeth of various animals, their modifications of form, and attachment to adapt them to modifications of function, and various classi- fications of the means of attachment have been made. Of these, perhaps the best and most logical is given by Charles Tomes in his Dental Anatomy, describing four forms of attachment: (1) By fibrous membrane; (2) by hinge-joint; (3) by ankylosis; (4) by insertion in a socket. These various forms of attachment will be taken up, and, if possible, the comparison between them and the evolution of the more complicated forms from the simpler will be shown. The study must begin with an examination of the structure and attachment of the placoid scales and the simplest form of tooth, as illustrated in the shark. Structure of Dermal Scales. — The dermal scales are composed of a conical cap of calcified tissue developed from within out- ward, by an epithelial organ, and corresponding in structure to the enamel. This cap is supported upon a conical papilla of calcified tissue formed from without inward, and corre- sponding to dentine. In the outer layer the arrangement of the fine tubules through the calcified matrix correspond very 272 THE ATTACHMENT OF THE TEETH closely to human dentine, but in the inner portions it is to be understood only by considering the formation of the dentine as progressing irregularly over the surface of the pulp and so dividing the pulp tissue into portions enclosed in large canals, from which the fine tubules radiate. The base of this partially calcified papilla has a calcified connective tissue built on to it by the derma or connective-tissue layer of the skin, which cor- responds to cementum forming the basal plate, spreading out Fig. 213 /M m Showing additions of bone of attachment to the bone of the jaw. (Tomes) more or less in the connective-tissue layer of the skin, and into which the fibers of this layer are built, so attaching the den- ticle or dermal scale to the deep layer of the coreum. This tissue very exactly resembles cementum. It is formed on the dentine as the cementum of a human tooth is, and shows the connective-tissue fibers embedded in it. In the ganoids the basal plates of adjoining scales unite, forming the armor plates of such fish as the sturgeon and gar-pike, and the dentical remains projecting from the surface of the plates. Attachment by Fibrous Membrane. — In the simplest teeth, as of the shark (Lamna cornubica. Fig. 3), which are typical ATT AC HM EXT BY HINGE JOINT 273 dermal scales, there is an exactly similar method of attach- ment, which may be taken as the simplest and most rudimen- tary, or attachment in a fibrous membrane. That is, there is no development or modification of the arch of the jaw, and the teeth have no direct attachment to the bone; in fact (Fig. 213), the jaws themselves are chiefly cartilage. Fig. 2U Attachment by hinge joint. Tooth of a hake: a, vasodentine ; h, pulp; c, elastic hinge; d, buttress to receive/, formed out of bone of attachment; e, bone of jaw; /, thickened base of tooth; g, enamel tip. (Tomes.) Attachment by Hinge Joint. — The formation of the hinge attachment as illustrated in many of the fishes (Fig. 214), may be understood as a modification of the attachment in a 18 274 THE ATTACHMENT OF THE TEETH fibrous membrane in a more highly specialized creature. These hinged teeth are found in many fishes and in the poison fangs of snakes. The jaws are calcified, and the basal plate or cementum may be considered as confined to, or specially developed on, one side of the dentine papilla, which is also more highly developed, especially in snakes. This cementum is built and calcified around the fibers of the fibrous tissue which pass directly to the bone of the jaw at that point. This bone is to be regarded as an addition to the jaw specially developed for each tooth. Thus, there is not only a modifica- tion in the arrangement of the cementum, but a development of bone for attachment of the tooth. The bloodvessels pass through the fibers of the hinge to the pulp, and are not affected by the motion of the tooth on the hinge; in fact, the pulp seems to be attached to the hinge. There are many complications of this method of attachment, but this may be taken as the type and the manner of its modification from the rudimentary conditions. The distinction, in this form of attachment, from the dermal scale consists in a modifica- tion of the arrangement of the cementum of the basal plate and a development of bone from the jaw to attach fibers which pass directly from cementum to bone. It should also be said that there are developments in the hinge teeth related to the third form of attachment, namely, ankylosis, which cannot be understood until this form is studied. Attachment by Ankylosis. — The third form of attachment, ankylosis (Fig. 215), or direct calcified union with the bone of the jaw, cannot be understood \\dthout a careful study of the nature and formation of the dentine in these rudimentary teeth. It is evident, from a study of the dentine of the dermal scales, that compared with human dentine, the tissue is rudimentary and not differentiated from other similar connective tissues. The tubules are comparatively very irregular, and resemble strikingly the tubules found in the secondary dentine formed by a degenerating pulp. The odontoblasts, or dentine-forming cells, are not like the highly specialized cells which form the primary human dentine, but resemble very closely simple spindle-shaped connective- ATTACHMENT BY ANKYLOSIS 275 tissue cells. The nucleus is larger and oval in form, and the protoplasm stretches off from it in one direction intQ a fibril Fig. 21; Tooth of scarus, showing attachment by ankylosis. (Owen.) 276 THE ATTACHMENT OF THE TEETH instead of in two directions into a spindle. The cells are much smaller than human odontoblasts and nearer the size of ordinary spindle cells of the human pulp. In fact, they look more like specially developed spindle cells than odonto- blasts. The formation of dentine begins on the surface, at the apex of a cone-shaped papilla of connective tissue, and proceeds inward. If the formation continues uniformly over the surface of the papilla, a solid layer of fine tubuled dentine results; but it often proceeds irregularly, apparently having special reference to the neighborhood of bloodvessels, so that irregular projections of dentine are found on its inner surface, dividing the pulp more or less into portions enclosed in larger channels or tubes. These may be very regular in arrangement and form around bloodvessels loops embedding the bloodvessel in the calcified tissue, producing what has been called vaso or vascular dentine; but the formation is still from the surface of the pulp until it is obliterated, except for what remains in the larger canals. As distinguished from this formation of dentine we find in the body of the dental papilla of many fishes the formation of spicules of calcified tissue, which resemble neither den- tine nor typical bone, shooting down through the substance of the pulp. They are more to be compared with the first formation of bone in membranes, or in the embryonal connective tissue of the body of the human jaw, which is afterward removed by absorption and replaced by true Haversian system bone. These calcifications contain lacunae, and have tubules or canaliculi running through them, and so, as Tomes says, are intermediate between dentine and bone. They divide the pulp into irregular spaces, and interdigitate, or perhaps actually join, the formation of dentine which has been progressing from the surface of the pulp. These spicules run down into the bone of the jaw, forming an actual calcified attachment for the tooth with the jaw; but in this view of it it is to be regarded as a calcification or rather a formation of bone in the pulp papilla interlocking with the dentine. In some of the fishes, as in Scarus, there is at the same time the remains of the cementum of the basal ATTACHMENT BY IMPLAXTATIOX IX SOCKET 277 plate formed on the outside of the dentine around the base of the cone. Ankylosis is confined to the teeth of many fishes, and may be stated as a modification from the dermal scale, resulting in the reduction or loss of the basal plate and an ossification of the pulp continuing through the connective tissue at the base of the pulp to the body of the jaw. Attachment by Implantation in Socket. — The development of the fourth form of attachment, by implantation in a socket, seems to be an evolution starting from the same point but proceeding in a different direction (Fig. 216). It Fig. 216 .-i, diagrams of transverse sections through the jaws of reptiles showing pleurc- dont (a), acrodont (6), and theodont (c) dentitions. B. a, lower jaw of Zootoca vivipara; b, of anguis fragilis. (After Leydig ) (Weidersheim, Comparative Anatomy of Vertebrates.) is associated with the very great increase in the size of the teeth and consequent necessity for a stronger attachment. The evolution of this is illustrated in the teeth of reptiles. Weidersheim classifies the teeth of reptiles as (1) resting upon a ledge on the lingual side of the jaw — pleurodont dentition; (2) resting on a slight ridge around them — acro- dont dentition; (3) lodged in permanent alveoli, as in the crocodile — theodont dentiton. These three classes illus- trate three stages in the development of the socket method of attachment. In the simplest form there is a cone-shaped tooth, attached 27S THE ATTACHMENT OF THE TEETH to the bone around its base, by the fibers being built into the cementum and bone. There is little modification of the rudimentary form, and little development of bone for its attachment. In a higher form the tooth has become long or peg-shaped, and the bone has grown up around a portion of it to support it; but it is attached to the bone by connective- tissue fibers, being built into the cementum on the surface of the tooth and into the bone of attachment on the jaw. The development of the form of the tooth to the peg from the cone may be understood as a continuing of the develop- ment of odontoblasts, and the formation of dentine (which always begins at the apex of the cone) farther and farther down the sides of the dental papillae. Then the formation of the cementum, which begins around the base of the cone and continues down on the outside of the calcified dentine, covering its outer surface, and building the connective-tissue fibers into the tooth. The development of bone accom- panies, or rather follows that of the tooth, building the other ends of these fibers into the bone which is developed to support the tooth. Summary. — To review the subject matter of this chapter, all teeth have been evolved from the simple placoid scale. In the simplest forms, as in the teeth of the shark, there is no relation to the bone whatever, but the fibers of the sub- cutaneous tissue are built into the basil plate of cementum. As the tooth becomes larger and demands more support, there is added to the bone of the jaw that which Tomes has called bone of attachment. The osteoblasts build up addi- tions to the jaw which surround and embed the fibers, so that the fibers which were originally in the subcutaneous tissue are fastened to the bone at one end and to the cementum at the other. The evolutions of attachment by hinge joint and by gomphosis are, therefore, direct evolutions from the simple attachment in membrane. The form of ankylosis is also evolved from the simplest type, but in this case the bone of attachment is associated with the pulp, and the formation of bone and dentine become interlocked and united. CHAPTER XXII THE PERIDENTAL MEMBRANE Ix one sense the peridental membrane may be consid- ered as the most important of the dental tissues, for upon it the usefulness of the teeth and their comfort to the indi- vidual is dependent. It makes no difference how perfect a crown may be, or how perfectly any damage which may have occurred to it may have been restored, unless the peridental membrane is in a healthy and fairly normal con- dition, the tooth will be useless, and the individual would be much more comfortable without it. Definition. — The peridental membrane may be defined as that tissue which fills the space between the surface of the root and the bony wall of its alveolus, surrounds the root occlusally from the border of the alveolus, and supports the gingivus. It is necessary to emphasize the three parts of the definition. The peridental membrane does not stop at the border of the bone, but continues to surround the root as far as the tissues are attached to it. In general, the dental profession has thought of the peridental membrane as only that tissue which occupies the space between the root and the wall of its alveolus. As will be seen from a study of a section later (Figs. 219 and 220), the structure of the tissue surround- ing the root between the gingival line and the border of the process is essentially the same as that in the alveolus, and quite difterent from the much coarser fibrous mat forming the submucous layer of the gum tissue. The peridental membrane also extends into the free margin of the gum and is the means of its support, holding the gingivae close to the surface of the tooth and supporting them in the interproximal spaces. The importance of this portion of the peridental 280 THE PERIDENTAL MEMBRANE membrane and the functions which it performs have been strongly emphasized in the last few years, in their relation to the extensions of caries and the beginnings of pyorrhea. Most of the diseases of the peridental membrane which result in the final loss of the teeth have their beginnings in this portion. Nomenclature.^ — The peridental membrane belongs to the class of fibrous membranes which form the covering of organs, the capsules of glands, and especially those membranes which cover the organs of support. Its closest relative is the periosteum in the attached portions, with which it has many points of structure in common, but it differs from the periosteum in any position in important respects. It has often been called the alveodental peri- osteum, but this name implies that the periosteum is folded down into the alveolus and back upon the surface of the root, which is an entirely erroneous conception of the mem- brane. This idea would imply that it was a double mem- brane having one layer covering the bone and another covering the root, the two uniting in the middle portions. But instead, the periosteum must be considered as stopping at the border of the alveolus,^ and being united with the peridental membrane around its circumference. ]\Iany writers use the word pericementum in place of peridental membrane. The author prefers and in this book will use the term peridental membrane, though the two are synon}'- mous. Divisions. — Purely for convenience in description, the peridental membrane is divided into three portions: The gingival portion, that portion of the membrane which sur- rounds the root occlu sally from the border of the alveolar process and supports the gingivae; the alveolar ])ortion, the portion of the membrane from the border of the process to the region of the apex of the root; and the apical portion, ' The student must be reminded that the word alveolus means a hole, and the alveolar process, the portion of the bone which contains the holes. In dental writing the word alveolus has often been incorrectly used in place of process or alveolar process. PLATE XI % V, V. 4' "^s -■^.■. ■ 'f> , i 4. i' ■■ ^i?**l*^iwirff Longitudinal Section of Peridental Membrane. Stained with heniatoxylin and eosin. Showing border of alveolar process. PLATE XI [ n:,i ^ '^ f • -ui \ i f J ,: t ^." \ '-S ii'^-'li^tfi.wia. -v.. te v§m Longitudinal Section of Peridental Membrane. Stained with hematoxylin and eosin. Showing part of the Ungual gingivus and border of the alveolar process. PLATE XIII % J 7 -J m S- 4'. - / .'-*'* ii4 H V / Transverse Section of Peridental Membrane. Stained with liematoxylin and eosin. Alveolar portion. FUNCTIONS 281 which surrounds the apex of the root and fills the apical space. These are illustrated in the diagram (Figs. 217 and 218). The Structural Elements. — These are: (1) White connective- tissue fibers; (2) fibroblasts; (3) cementoblasts; (4) osteo- blasts; (5) osteoclasts; (6) epithelial structures which have sometimes been called the glands of the peridental membrane; (7) bloodvessels; and (8) nerves. Fig. 217 Drawing to show the arrangement of the fibers in a labiolingiial section through an incisor of a kitten. (Black ) Functions. — ^The peridental membrane performs three functions: (1) A ph^-sical function — it maintains the tooth in relation to the adjacent hard and soft tissues. (2) A vital function — the formation of bone on the alveolar wall and of cementum on the surface of the root. (3) A sensory function — the sensation of touch for the tooth being exclu- sively in this membrane. It is necessary to emphasize the two parts of the physical 282 THE PERIDENTAL MEMBRANE Fig. 218 G -! Al Ap •! Diagram of the fibers of the peridental membrane: G, gingival portion; Al, alveolar portion; Ap, apical portion. (From a photograph of a section from incisor of sheep.) ARRANGEMENT 283 function; the peridental membrane not only supports the teeth in their relation to the bones which carry them, and sustains them against the forces of occlusion and masti- cation, but it also sustains the soft tissues in their proper relation to the teeth. The second part of the physical function is fully as important as the first, and the study of the structure of the tissue related to it and the adaptation of the form of the gingivse to the anatomic form of the teeth and alveolar process, are important considerations which should never be lost sight of in the making of artificial crowns. Classes of Fibrous Tissue. — The fibrous tissue of the peridental membrane is entirely' of the white variety, but may be divided into two classes. The principal fibers and the indifferent or interstitial tissue. The former perform the physical function of the membrane, the latter simply fill in spaces between the bundles of fibers and surround and accompany the bloodvessels and the nerves. The Principal Fibers of the Peridental Membrane. — These may be defined as the fibers which, springing from the cementum, are attached at their other extremities to the connective tissue supporting the epithelium, the fibrous mat of the gum tissue, the cementum of the approximating tooth, the outer layer of the periosteum at the border of the alveolar process, or the bone of the alveolar wall. Arrangement.— The principal fibers literally spring from the cementum, the cementoblasts building up the matrix around them and then calcifying both the matrix and the fibers, in this way attaching them to the surface of the root. In most places the fibers as they spring from the cementum appear as good-sized bundles. A short distance from the surface of the root they may break up into smaller bundles which anastomose and interlace, passing around bloodvessels and other fibers in their course and being again united into large bundles for attachment at their other extremity. To arrive at an understanding of the arrangement of the fibers of the peridental membrane, sections must be cut longitudinally, both from buccal to lingual and from mesial 2S4 TIIE PKRJJJKXTAL MEMBRANE to distal, and transversely through all portions of the mem- brane. It therefore re(iuires the study of many sections to work out a complete conception. After studying them out completely in this way one is impressed with the beautiful adaptation of their arrangement to sustain the tooth against all the forces to which it is subjected, and to support the free margin of the gum, so that it will lie closely against the gingival portion of the enamel. It is necessary, however, to remind the student that connective tissues are formed in response to mechanical conditions and stimuli, and therefore this arrangement must be considered, not as having been designed to sustain the forces, but as being the result of the forces to be sustained, and therefore beautifully adapted to them. Beginning at the gingival line, the fibers springing from the cementum pass out at a short distance at right angles to its surface and then bend sharply to the occlusal, passing up into the gingivus and uniting with the fibrous mat which supports the epithelium. These are much more strongly marked on the Ungual than on the labial gingivus, because in mastication the lingual gingivus receives more pressure of food, which would tend to crush it down. A little deeper the fibers springing from the cementum on the labial and lingual pass out at right angles to the cementum and are lost in the coarser fibrous-mat of the gum tissue. The distance which they extend before lost in the coarser fibers is always greater on the lingual than on the labial. On the proximal sides the fibers springing from the cementum at the same level, branch and interlace, passing across the interproximal space, to be attached to the cementum of the approximating tooth. These fibers are of the greatest importance, as they produce the basket work which forms the supporting framework for the interproximal gingivus. A little farther occlusally the fibers as they come from the cementum are inclined apically. A short distance from the cementum they unite into very large and strong bundles which join with the fibers of the outer layer of the perios- teum, extending over the labial and lingual border of the ARRAXGEMEXT 285 alveolar process. On the proximal sides the fibers at the same level are attached to the cementum of the adjoining tooth, or are inclined apically, to be inserted in the bone of the septum. These large bundles form a distinct la^'er, which has been called the dental ligament, and bind them together across the septum. They are the only fibers which hold the teeth down in its socket. At the border of the alveolar process, and in the occlusal third of the alveolar portion, the fibers pass directly from the cementum to the bone at right angles to the axis of the tooth. In this position the fibers are larger and stronger, and show less tendency to break up into smaller bundles in their course than in any other portion of the membrane. In the middle and apical thirds of the alveolar portion the fibers are inclined occlusally as they pass from the cementum to the bone. They spring from the cementum in compact bundles, and show a strong tendency to break up into fan-shaped fasciculi, spreading out as they approach the bone, to be attached over a larger area of the alveolar wall. These fibers literally swing the tooth in its socket and support it against the forces of mastication. In the apical region fibers springing from the cementum pass out in all directions, spreading out in the same way, to be inserted into the bone forming the wall of the apical space. If force is exer^d against the lingual surface of an incisor, the fibers on the lingual side of the root in the occlusal third will sustain part of the strain, preventing the cro\NTi from moving labially, and at the same time the fibers on the labial side of the root in the apical space will also be under strain, preventing the apex of the root from moving lingually. The general plan of arrangemenit which has been described is illustrated in Dr. Black's diagram made from a labio- lingual section of an incisor of a young kitten (Fig. 217). With this general plane of arrangement in mind individual sections may be studied, examining the arrangement and appearance of the fibers in detail. Figs. 219 and 220 show the labial and lingual gingivae from an incisor of a sheep. Notice that the labial gingivus is taller and thinner, and the 286 THE PERIDENTAL MEMBRANE fibers passing up into it are not as strongly marked. Notice also the distance to which the final fibers of the peridental membrane can be followed before they are lost in the coarser mat of gum tissue. The lingual gingivus is broader and flatter, and the fibers passing up into it form a strong Fia. 219 •\^^ Longitudinal section of the peridental membrane in the gingival portion, from a lamb (the labial gingivus). and well-defined band. Under higher magnification, fibers would be seen cut transversely in the gingivus, which pass around the tooth, helping to hold it closely against the enamel. In Fig. 221 the fibers uniting with the outer ARRAXGEMENT 287 layers of the periosteum are very well shown. Taking transverse sections in the gingival portion and remem- bering that they are cut at right angles to these through the same area, the distribution of the tissues will be bet- ter understood. Fig. 222 shows a section cut close to the Fig. 220 Longitudinal section of the peridental membrane in the gingival portion (the lingual gingivus): D, dentine; N, Nasmyth's membrane; C, cementum; F, fibers sup- porting the gingivus; Fi, fibers attached to the outer layer of the periosteum over the alveolar process; F-, fibers attached to the bone at the rim of the alveolus; B, bone. (About 30 X) gingival line. At A the epithelium on the labial surface of the gingivus is seen, and at B the epithelium lining the gingival space. On the proximal sides of the roots the fibers will be seen passing from the cementum of one tooth to that of the next. Fig. 223 is a httle deeper and shows the fibers Fig. 221 Longitudinal section of peridental membrane of young sheep, showing fibers penetrating the cementum: D, dentine; C, cementum, showing embedded fibers; F, fibers running to the outer layer of the periosteum, covering the aveolaf process; F', fibers running to the bone at the border of the process; B, bone. (About 80 X) ARRANGEMENT 289 attached around the entire circumference of the root. Begin- ning at the middle of the labial surface, the fibers will be found springing from the cementum and passing out at right angles to it, to be lost in the fibrous mat supporting the epithelium. The fine fibers of the peridental membrane can be followed for about half the distance to the epithelium before they are lost in the coarser mat of gum tissue, and a fairlv definite boundary will be seen between what should Transverse section of the peridental membrane in the gingival portion, from young sheep. The roots of two temporarj' incisors are cut across. The epitheUum lining the gingival space is shown part way around one. A, epitheUum on labial surface of gingivae; B, epithelium lining the gingival space. (About 60 X) be considered peridental membrane and the gum tissue. As the distolabial angle of the root is approached, the fibers passing from the cementum ten.d to swing around distally, and pass to the mesiolabial angle of the adjoining tooth. Along the proximal surface the network which supports the interproximal gingivus is well shown. The fibers springing from the cementum interlace and pass around bloodvessels and fibers which are passing up into the gingivus, and finally 19 290 THE PERIDENTAL MEMBRANE are inserted into the cementiim of the next tooth. In this way it will l^e seen that the teeth in the entire arch are firmly bound together by the fibers in the gingival portion. This Fig. 223 Transverse section of the peridental membrane in the gingival portion (from sheep): E, epithelium; F, fibrous tissue of gum; B, point where peridental mem- brane fibers are lost in fibrous mat of the gum; P, pulp; F' , fibers extending from tooth to tooth. (About 30 X ) explains the way in which the positions of all the teeth are affected by the loss of a single one in the arch, and the way in which the movement of one tooth will draw its neighbors after it. It also explains the separation of the central PLATE XiV Transverse Section of the Peridental Membrane in the Occlusal Third of the Alveolar Portion (from Sheep). .V, muscle fibers; Per, periosteum; .4/, bone of the alveolar process; Pd, peri- dei-ital nien-ibrane fibers; P, pulp; D. dentine; Cm, eementum. ARRANGEMENT 291 incisors when the freniim labium passes through between the teeth, and is inserted on the lingual surface of the alveolar process. If these incisors are to be held together perma- nently, normal attachment of fibers extending from the cementum of one tooth to that of the other must be secured. The fibers in this area are also well shown in Fig. 224, and Fia. 224 ■'/-'. ** s A portion of the peridental membrane between two incisors of a young sheep, showing the fibers extending from tooth to tooth. it can be understood how they form foundation upon which the interproximal gingivus rests. The first step in the sagging of the interproximal gum tissue is the cutting off of the fibers from the cementum, where it bends occlusally, following the curve of the gingival line on the proximal surface. Plate XIV shows a transverse section in the occlusal third 292 THE PERIDENTAL MEMBRANE FiQ. 225 ■«^^- y.-'.j i -uv W^ ^^^!^fA m^^ . 0% ^^4^^MP^^^K^^^9|H^^^H^^^jN^ t % '-CV" »""* 3 >« < Diagram of peridental membrane from section similar to Fig. 224. (From Maloc- clusion of the Teeth, Dr. E. H. Angle.) Fig. 22G Fibers at the border of the alveolar process (from sheep): D, dentine; C, cementum; F, fibers extending from cementum to bone; Bl, bloodvessel; B, bone. (About 80 X) A RRA N GEM EN T 293 of the alveolar portion from the incisor of a sheep. Upon the labial a few muscle fibers are seen and the periosteum covering the labial surface of the process. Notice the medul- lary spaces in the bone and the canals opening into the peridental membrane and periosteum. The light line forming the outer boundary of the dentine is characteristic. Two layers of cement um are seen, and notice the thickening of the layer where strong bundles are attached. At the middle of the labial surface the fibers pass at right angles to the cemen- tum and are attached to the bone, but as the distolabial angle of the root is approached the bundles swing distally to be attached in the bone. In Fig. 225, which was drawn very carefully from this section, the arrangement of the fibers is shown diagrammatically. Notice the way in which they pass over and under each other and around the bloodvessels which wind through them. This relation to the bloodvessels is important, and will be considered again later in connection with the blood supply of the membrane. The tangential fibers at the angle of the root hold the tooth against the forces which tend to rotate it in its socket. They are impor- tant in connection with all rotating movements in ortho- dontia. It has long been noted that rotations were the hardest movements to retain, especially if the tooth were moved in no other direction. In this case, if the tooth were turned mesially the fibers at the distolabial angle would spring the thin plate of the alveolar process as a bow is bent, leaving a condition of stress in the tissue which will tend to spring back into its old position and drag the tooth with it. Notice the greater thickness of the membrane on the lingual as compared with the labial. Figs. 221 and 226 show longitudinal sections at the border of the alveolar process. Notice that the fibers can be seen running through the entire thickness of the cementum. They are large, strong fibers and branch \ery little in their course. Note the bloodvessel that is shown in several of these sections, and the way in which it gives oft' branches passing over the border of the processes and toward the cementum. CHAPTER XXIII THE CELLULAR ELEMENTS OF THE PERIDENTAL MEMBRANE Fibroblasts. — The fibroblasts are found everywhere between the fibers which they have formed and to which they belong. They are spindle-shaped or stellate connective-tissue cells, having a more or less flattened nucleus and a body of granu- Fig. 227 Fibers and fibroblasts from transverse section of membrane: F, fibers cut trans- versely; F^, fibers cut longitudinally, showing fibroblasts. (About 80 X) lar cytoplasm, which is squeezed out into thin projections between the fibers. In sections stained with hematoxylin the cells take the stain strongly and the fibers remain clear (Fig. 227). In this way the fibers are marked out by the cells which lie between them. The number of the fibroblasts CEMEXTOBLASTS 295 in the membrane decreases with age. They are large and numerous in the membrane of a newly erupted tooth and are comparatively small and few in the membrane around an old tooth. This is, however, characteristic of fibroblasts in connective tissue generally. Fig. 227 shows a small field taken from the gingival portion of the membrane between the teeth. The magnification is low, the photograph being made with a f objective. The cells are seen as little dark dots lying between the fibers, which are clear. Where the fibers are cut longitudinally they appear spindle-shaped, but where the fibers are cut across they appear star-shaped. They will be seen better in photographs made with higher magnification, but an adequate idea of their form can only be obtained by studying sections very carefully with a J or -jV objective and using the fine adjustment to gain an idea of the third dimension of space. They are shown in many of the illustrations of the epithelial structures. Cementoblasts. — The cementoblasts are the cells which form cementum. They cover the surface of the root every- where between the fibers which are embedded in the tissue. While these cells perform the same function for the cementum as the osteoblasts do for bone, they are quite dift'erent in form. They are always flattened cells, sometimes almost scale-like, and when seen from above very irregular in out- line. This irregularity in outline is due to the projections of the cytoplasm around the fibers as they spring from the cementum, the edges of the cell being notched and scalloped to fit about them. There is a central mass of granular cytoplasm which contains an oval and more or less flattened nucleus, from which the cytoplasm extends in projections passing partly around the fibers. Isolated cementoblasts are shown in Fig. 228, drawn by Dr. Black. In order to obtain an idea of the form of the cementoblasts, sections must be cut at a tangent to the surface of the root, and just miss- ing the surface of the cementum. In this way the fibers are cut across and the cementoblasts are shown covering the entire surface between the fibers. These are shown in Fig. 229, in which the fiUers are left perfectly clear in order to 296 THE PERIDENTAL MEMBRANE outline the cells more distinctly. In sections cut at right angles to the surface of the roots (Figs. 240, 241, and 242) the cementoblasts are shown as more or less flattened, but no idea of the way in which they fit about the fibers can be obtained. Cytoplasmic processes extend from the body of the cemen- toblasts into the matrix of the cementum. These correspond to the processes of the osteoblasts which occupy the canal- iculi of bone. . They, however, are not nearly as numerous or as regular in their arrangement as the osteoblasts. Pro- cesses extending from these cells in a direction from the cementum out into the tissue of the membrane have not been demonstrated. Fig. 22S 4 ^ t^^\^ Isolated cemeotoblasts. showing the form of the cell as it fits around the fibers springing from the cementum. >■■■'■ Fig. 229 Cementoblasts as seen in a section at a tangent to the root and just miss- ing the cementum. The fibers are left white, the ceils are shaded. Cement Corpuscles. — Occasionally a cementoblast becomes fastened down to the surface and enclosed in the matrix that is formed. The}^ then lie in a lacuna and show processes radiating from them into the canaliculi. These correspond to bone corpuscles, but there is no such regularity of their disposition or arrangement with reference to the lamellae, as is shown in the case of bone. In man the cementum in the gingi\'al half of the root is usually without cement corpuscles. They often lie entirely within a single lamella instead of OSTEOBLASTS 297 between two, as is the case in bone. In general they are found where the layers are thick and the embedded fibers are not specially numerous. They are very often seen where absorptions have been refilled by the formation of subse- quent layers (Figs. 15-1 and 155). It is by the activity of the cementoblasts producing a new layer of cementum that the fibers are attached to the surface of the root. In studying many sections, places are found where the fibers, though lying in contact with the surface are not attached to the cementum. In some places it can be seen that they have been cut off by absorptions. From a study of these layers it is evident that there is a constant readjustment in the attachment of the fibers to the root during the function of the tooth, which probably adapt it to slight changes of position resulting from wear and other conditions. It is important to remember that whenever the fibers have been stripped from the surface of the cementum, they can be reattached to it only by the formation of a new layer of cementum, building the fibers into it. This is certainly possible if the conditions are properly controlled, but the cells of the tissue must be in a normal and vitally active condition, and the surface of the root must be such that they can lie in physiological contact with it. The cure of a pyorrhea case, therefore, becomes a biological problem. In this connection it is important to remember that a surface of cementum which has long been bathed in pus may be so filled ^\'ith poison that no cell can lie in contact with it and perform its functions. Osteoblasts. — The osteoblasts of the peridental membrane are exactly like osteoblasts in other positions. They cover the surface of the bone of the alveolar wall lying between the fibers which are embedded in it. Even in the young subject they are not found in every position, while in an adjoining area the surface of the bone may be covered with them. In the old subject they are generally absent or have been reduced to flattened scales, which are very difficult to demonstrate; but even in these cases areas will be found in which osteoblasts are present. These are areas of active 298 THE PERIDENTAL MEMBRANE bone formation. The osteoblasts lay down bone exactly as occurs in attached portions of the periosteum, but after Fig. 230 PdM PdB Penetrating fibers in hone. A field from jjlute X\': Pil M, peridental membrane; 0¥, osteoblasts of peridental membrane; Ob^, osteoblasts of medullary space; Pd.B, solid subperidental and subperiosteal bone with embedded fibers; Ms, medullary space formed by absorption of the solid subperidental bone with embedded fibers; H.B, Haversian system bone without fibers built around the medullary space, (About 200 X) HB PLATE XV HB PdB Border of Growing Process. Cm, eementum; Pd, peridental mennbrane; Pd. B, solid subperidental and subperiosteal bone with enmbedded fibers; Ms, medullary space Ibrnned by absorption of the solid bone; H. B, Haversian systeni bone without fibers; Per, periosteuni. (About SO X) OSTEOCLASTS 299 a little thickness of this solid peridental bone has been formed it is perforated by penetrating canals, on the walls of. which absorptions occur, forming spaces about which new Haversian system bone is formed. This is illustrated in Plate XV. In this way only sufficient subperidental bone is left to furnish an attachment for the fibers. Fig. 230 shows a higher magnification of a small area. The osteoblasts are seen between the fibers on the surface of the alveolus, and the fibers can be followed through the subperidental bone. A large absorption area has been formed which has been partly rebuilt, and the new-formed bone without embedded fibers is lighter in color. An under- standing of this building and rebuilding of bone through the agency of the peridental membrane is necessary to understand the development of the face and everything in connection with tooth movement, whether physiological or artificial. Osteoclasts. — The osteoclasts of the peridental membrane are not constant elements. They appear and disappear in response to the same conditions which lead to their appear- ance and disappearance in bone. They are always large, multinuclear cells, having from three or four to thirty or forty nuclei (Fig. 231). They may appear upon the surface of the cementum, upon the surface of the alveolar wall, or within the medullary spaces of the bone. They are formed from embryonal cells in the tissue in response to mechanical stimuli. ^Morphologically they are in no respect difterent from the osteoclasts in bone. The osteoclasts are tissue destroyers and are the active agents in the removal of any hard tissue. There is no difi'er- ence in them, whether they are destroying the fibrous tissue, bone, cementum, or dentine (Fig. 232). In order for them to act, their cytoplasm must lie in actual contact with the surface to be attacked. They do not first decalcify and then remove, but apparently by applying their cytoplasm to its surfacejthe cells destroy the intercellular substance, forming hollows in the surface, into which the cells sink. These hollows have been called Howship's lacunae. The cells 300 THE PERIDENTAL MEMBRANE usually appear in groups and spread out over the bone or cementum to be attacked, but sometimes only two or three will be found at a point on the surface of the bone, and these will burrow into the substance, forming a penetrating canal Fig. 231 Osteoclast absorption of bone over permanent tooth; Oc, osteoclasts; B, bone of crypt wall; F, fibrous tissue of follicle wall; -4, ameloblasts. (About 62 X) running through the bone (Figs. 233 and 234). In these positions the osteoclasts are usually comparatively small. As fast as the canal is formed the embryonal cells of the mem- brane multiply and grow into the space and at any point OSTEOCLASTS 301 where absorption is going on the portion destroyed is immedi ately replaced by embryonal connective tissue. Fig. 232 Osteoclasts in cancellous bone near the peridental membrane; in some portions of the field osteoblasts are seen. As bone is removed note how embryonal connective tissue replaces it. This will be noted in all the illustrations showing absorp- tions. Whenever absorption is going on formation is also 302 THE PERIDENTAL MEMBRANE going on in an adjoining area. In this way the function of the tissue is maintained until the last remnants of it are destroyed. The general statement may be made that bone formation is always accompanied by bone destruction, and bone destruction by rebuilding. The result depends upon which side the balance swings. The alternation of formation and absorption in the removal of hard tissues is well illus- trated in the absorption of the roots of the temporary teeth. Fig. 233 Osteoclast absorption forming penetrators of canal: a, bone matrix; h, bloodves- sel; c, embryonal connective tissue; d, new bone formation; e, osteoblasts, /, osteo- clasts. (Black.) The absorption does not begin at one point and spread continuously over the entire surface of the root. If it did so, all of the fibers would be cut out and the tooth would drop off with at least a considerable portion of the root. The process progresses in something of this fashion. At a point on the side of the root near the apex, where the growth of the erupting tooth produces pressure, osteoclasts appear in the membrane, cutting off the fibers, displacing the cemen- toblasts, and arranging themselves in groups on the surface OSTEOCLASTS 303 of the root. These dissolve away the cementum and sink into the tissue, perhaps cutting into the dentine for a short distance. By this excavation the pressure is relieved, the osteoclasts disappear, cementoblasts are formed in the Fig. 234 A longitudinal section through the remains of the alveolar process around the root of a temporary tooth about to be shed (sheep): C, the cementum on the remains of the tooth; B, penetrating canals cut through the labial plate of bone. embryonal connective tissue, and the deposit of cementum begins in the excavation, reattaching the fibers in this area. As the rebuilding progresses, at a point a little farther occlu- Fig. 235 Root of a temporary incisor, sho-wing absorption and rebuilding of cementum (from sheep): G, gingivus; D, dentine; Cm, cementum; Ab, absorption cavity, showing Howship's lacunae; Cm'^, new-formed cementum. (About 50 X) OSTEOCLASTS 305 sally, osteoclasts appear and begin a new excavation. In this manner the process continues. When the absorption stops in the second point, it begins again at the first, cutting Fig. 236 A transverse section through an incisor from the same jaw as Fig. 235, and at the level of Cm^, showing the refilling of the absorption cavity by new layers of cementum. much deeper into the dentine, and, oscillating back and forth, it progresses until all of the dentine may be destroyed, leav- ing the hollow cap of enamel, and even then new-forming 20 306 THE PERIDENTAL MEMBRANE cementum to maintain the attachment will be fomid around the circumference. In this way it will be seen that the function of the tooth is maintained until its successor is ready to take its place in a very short time. The importance of this arrangement will be more fully appreciated after a study of the relation of the teeth to the development of the face. Fig. 235 shows a longitudinal section through a temporary incisor of a sheep. At Ah an absorption has just been completed, for the osteoclasts have disappeared. The excavation is seen filled with embryonal tissue and rebuilding is about to begin. At Cm an older and much larger absorption space is seen which has been partially replaced by a formation of new cementum reattaching fibers. In Fig. 23G a transverse section of the root is seen which is from the same jaw cut at the level of Cm, and shows the absorption refilled. This patchwork performance goes on in the same way in the bone of the alveolar process, and its study is one of the most interesting phases of the rela- tion of the teeth to the development of the face. Without a clear idea of this it is impossible to understand how the teeth, after their roots are fully formed, can move through three dimensions of space and retain their function all the time. Epithelial Structures.— The epithelial structures of the peridental membrane were first described by Dr. Black in his volume Periosteum and Peridental Memhrane, pub- lished in 1887. At this time Dr. Black considered them to be of lymphatic character and named them endolymphatics. His conception of them was that they were lymphatic channels crowded with adenoid cells. Since then the form and appearance of the cells and the character of their reaction with staining agents has shown the cells to be of epithelial character. In the same year that Dr. Black's book was published, von Brunn^ described the same struc- tures. He considered them as epithelial remains of the outer layer of the enamel organ, growing down around the 'ArcLiv f. Anatomie, 1887. DISTRIBUTION 307 root beyond the gingival line where the formation of enamel stops. It has seemed probable to the writer that this was correct, but their histogenesis has not been sufficiently well followed, and it presents an attractive field for research. Although this is the origin of these structures, it has never seemed proper to regard them as embryonal remains, for while, like all the cellular elements, they are more numerous in young people than in old, they are persistent throughout life. They have been shown in the membrane from a man aged seventy years, and it does not seem logical to suppose that embryonal debris that was useless to the organism would persist through life. Up to the present time, however, nothing has been discovered about these structures to throw any light upon their function. Specimens have strongly indicated that they were important in some pathologic conditions. Their cells have been found dead and degener- ating in pathologic material beyond the point showing any pathologic condition in other cells. These structures have been observed in sections from man, sheep, cat, dog, and monkey. The best material for their study is a young sheep or pig. Distribution. — These structures are composed of cords or rows of epithelial cells, surrounded by an extremely delicate basement membrane (Fig. 2.37). In some cases there is a slight indication of a circular arrangement of connective tissue around them. The cords lie very close to the surface of the cementum, winding in and out anjong the fibers (Fig. 238). They anastomose and join with each other, forming a network the meshes of which are comparatively close in the gingival portion (Fig. 239), and comparatively wide in the apical portion, the cords becoming scarcer as the apex of the root is approached, but the author has seen them in sections from the apical third. A binocular microscope was used to obtain a true concep- tion of the way in which these cords wind in and out among the bundles of fibers. The cords show a marked tendency to run out into the membrane and loop back (Fig. 240), coming very close to the surface of the cementum. 308 THE PERIDENTAL MEMBRANE The ends of the loops toward the cementum often show enlargements which in some cases apparently lie directly in contact with the cementum (Figs. 241 and 242). These enlargements next to the cementum are shown in Fig. 240. The Arrangement of the Cells. — There is no definite arrange- ment of the cells in these cords. In some places there will be a ring of irregular polyhedral or rounded cells which almost exactly resemble a simple tubular gland. In other places there is a pretty definite outer ring of cells and a central Fig. 237 ^s^ M Diagram of glands of peridental membrane. (Black.) mass enclosed by them. The cells are made up of granular cytoplasm, each containing an oxoid nucleus that is rich in chromatin. The author has spent much time attempting to work out the relation of these cords to the epithelium lining the gingival space, thinking that possibly they open into it. In a few places structures appearing very much like a duct have been seen, as shown in Fig. 244, but they are apparently only unusually large cords. There is no regularity in places where they are found, and no con- A sectioEL cutting diagonally through the root, showing the network of epithelial cords, A; dentine, D; cementum, Cm. 310 THE PERIDENTAL MEMBRANE nection with the gingival space has ever been discovered. Toward the gingival, as the gingival line is approached, the cords seem to swing out away from the cementum, espe- cially on the proximal side, and to pass up into the gingivus, where they are lost among the projections of the epithelium. Fig. 239 Transverse section of the perideni the position of the epithelial cords. in Fig. 241 is seen. il membrane in the gingival portion, showing At 1 the loop shown in higher magnification Gland of Serres. — Salter, in his Dental Pathology and Sur- gery, quotes Serres, who assigns the function of a gland to the epithelium lining the gingival space. This, the writer believes, is the first reference to an appearance in the tissues that has been called the gland of Serres. It has long been noted that the epithelium lining the gingival space was lighter in struc- ture, composed of larger cells, and had no horny layer on its GLAND OF SERRES 311 surface, as is true of the epithelium on the outer surface of the gingivus. Upon the proximal surfaces the projections Fig. 240 Epithelial structures of the peridental membrane (from sheep): Fb, fibroblasts; Ec, epithelial structures; Cb, cementoblasts; Cm, cementum; D, dentine. (About 468 X) of the epithelium which extend down between the papilla^ of connective tissue, which constitute the stratum papillaris, are specially long, and in the connective tissue between 312 THE PERIDENTAL MEMBRANE them collections of small round cells are often found. It is between these projections of epithelium that the cords of epithelial cells which have been described are lost, and to this portion of the tissue Dr. Black has again called attention, Fig. 241 Epithelial structures: Ec, epithelial cord, apparently showing a lumen; Ch, cemento- blasts; Cm, cementum; D, dentine. This loop is seen in Fig. 226. as the gland of Serres. Sufficient work has not xo^t been done upon this subject to know whether this is a constant arrangement, or whether it is found only in certain animals, or even whether it may not possib'y be pathologic. The appearance is shown in Plate XVI and Figs. 245 and 246. BLOODVESSELS 313 Bloodvessels. — The peridental membrane possesses a very rich blood supply. A number of vessels enter the membrane in the apical portion from the medullary spaces in the bone. Fig. 242 Transverse section, showing the cellular elements: Fb, fibroblasts; Ec, epithelial structures: Cb, cementoblasts; Cm, cementum; D, dentine, (About 900 X) Some of these, passing through canals in the apex of the root, supply the dental pulp, others pass up through the mem- brane. As they extend occlusally they give off and receive branches which enter the membrane from the bone of the :U4 THE PERIDENTAL MEMBRANE alveolar wall. In this way the caliber of the principal vessels is maintained throughout their course in the membrane. As they reach the border of the alveolar process they give oflF Fig. 243 Epithelial structures (from sheep): /'7), fibrohhists; /w, ei)itheHal structures; Cb, cementoblasts; Cm, cementuni; D, dentine. (About 700 X) branches which anastomose with the vessels of the perios- teum and gum tissue. These are shown in Plate XVII. In the young membrane these vessels occupy a position A m: Wp^^^ \ I r > X < "^ ^^ ' BLOODVESSELS 315 closer to the bone than the cementiim, and as the membrane becomes thinner they often come to He in grooves in the bone. Vessels of any size are rarely seen close to the cementum, and the capillaries in the membrane are rather scarce, though they are more numerous than in most connective tissues of as compact a character. The anastomosis of the vessels in the membrane is quite rich. It is important to remember Fin. 244 A very large cord which was at first mistaken for a duct. that the cancellous bone of the process is richly supplied with bloodvessels, and the anastomosis with the vessels of the membrane, from the alveolar wall and over the border of the process, is important in the consideration of pathologic conditions. In alveolar abscess the vessels entering through the apical space ma}' be entirely cut ofl', but this does not disturb the blood supply of the rest of the membrane. The removal of the pulp has often been advocated in the treatment 316 THE PERIDENTAL MEMBRANE Fia. 245 Longitudinal section, cut mesiodistally, similar to Plate XVI: D, dentine; Cm, cementum which has separated from the dentine; Gs, gingival space; Ep, epithelial projection from the lining of the gingival space, Ec, epithelial cords; Rs, small round cells in the connective tissue. BLOODVESSELS 317 Fig. 24G A longitudinal section cut mesiodistally: E, epithelium of the gingivus; Gs, gingivs space; Cm, cementum -which has separated from the dentine; Ec, epithelial cords. 318 THE PERIDENTAL MEMBRANE of pathologic conditions of the membrane, on the ground that the vessels entering the pulp rob the membrane of blood supply, and that their removal made recovery more certain. No one having a knowledge of the blood supply of the membrane could advise this for this reason. In their course through the membrane the vessels wind between the principal fibers in a way that can only be appreci- ated by studying sections with a binocular instrument, and when this condition is realized it can be understood how some inflammations in the membrane are set up. For instance, when force is applied to a tooth, the principal fibers are stretched. This causes them to close some spaces and open others. The vessels in the closed spaces are constricted and the flow of blood through them partly shut off. The vessels in the enlarged spaces dilate to compensate. If the force is removed, the dilated vessels are again constricted, and the constricted ones enlarged, and the result is a literal sawing upon the walls of the bloodvessels which in a very short time will set up an acute inflammation. This is extremely important in the application of force in orthodontia, and often also in the use of the mallet in condensing gold, especially for young patients. Nerves. — The nerves of the peridental membrane enter the peridental membrane in company with the bloodvessels. Their source is the same as that of the bloodvessels. The trunks entering in the apical space contain from eight or ten to fifteen or twenty medullated fibers. Some of these enter the dental pulp, others extend up through the mem- brane, winding in and out among the fibers, generally following the course of the bloodvessels. Many trunks containing eight or ten fibers enter through the alveolar wall. In this way a fairly rich plexus is formed, from which fibers are continually given off to be lost in the tissue. They probably terminate in beaded free endings. No special nerve endings have been demonstrated. A few Pacinian corpuscles have been seen near the gingival border. These are not generally found, however. The nerves of the mem- brane give to it the sense of touch, which is the only sensory CHANGES IX PERIDENTAL MEMBRANE WITH AGE 319 function of the membrane. As has been noted in connec- tion with the dental pulp, the hard tissues and the pulp have no sense of touch. The contact of any substance with the surface of the tooth is reported to consciousness through the medium of the peridental membrane. For instance, the slightest touch of a delicate instrument produces a slight movement of the tooth which affects the nerves between the- fibers. The delicacy of this mechanism can be demon- strated by the following experiment. Lightly touch the surface of the enamel and the patient will tell at once not only which tooth is touched, but whether a steel instrument or a wooden point or some soft material was used. If, however, the finger is placed upon a surface of the tooth and firm pressure made in one direction, the contact of the point will not be recognized. The Changes in the Peridental Membrane with Age. — The teeth are formed in crypts in the bone, and when they begin to erupt the roof of the crypt is removed by absorption, making an opening large enough for the crown to pass. As the root is formed, the tooth moves occlusally and the alveolus grows up around it, beginning at the margins of the crypt. When the tooth first erupts, therefore, the alveolus is much larger than the root, and the fibers of the peridental membrane are very long. The size of the alveolus is reduced by the formation of bone, by the osteoblasts on its wall, and the size of the root is increased by the formation, layer after layer, on its surface. In this way the thickness of the mem- brane is reduced. Figs. 247 and 248 were made to illustrate this change. They were photographed with as nearly the same magnifications as possible, so as to compare the thick- ness of the layers. In the first, there are but two layers of cementum; notice the thickening of the last-formed layer, to attach the strong bundles of fibers at the angle of the root. The second is from a temporary tooth which has been in position and function for a long time; notice the thickness of the cementum and that the formation of bone and cemen- tum has reduced the thickness of the membrane to not more than one-third of its original amount. Notice also that the 320 THE PERIDENTAL MEMBRANE surface of both bone and cementum are not even, but scalloped, and that where the cementum projects toward the alveolar wall there is a depression in the bone, and where the bone projects toward the cementum there is a depression in the cementum. There is, therefore, a distinct tendency Young membranes (from sheep): D, dentine; Cm, cementum; C'mi, thickening of cementum to attach fibers at the corner; Pd, peridental membrane; B, bone forming the wall of the alveolus, (About 80 X ) for the two tissues to interlock but remain separated by a layer of fibrous tissue. The author has never seen a specimen showing a union between calcified substances of bone and cementum. Two surfaces of cementum may become united by direct calcification and the teeth fused together. This PRACTICAL CONSIDERATION 321 is illustrated in many freak specimens to be found in any dental museum. It is often stated that a tooth had become ankylosed to the bone, but to the author's knowledge no specimen has ever been shown in which the separating layer of fibrous tissue was not present. Fig. 248 Old membranes (from sheepi: D, dentiue: Cm, cemeuiuiu; Pd, pendeutal mem- brane; 5, bone forming the wall of the alveolus; P, pulp. (About 80X) Practical Consideration. — These structural facts are of the greatest practical importance, especially in the making of gold fillings for young persons. Every operator has noticed the greatest difference in the feeling of the instrument under the mallet upon difterent teeth. In one instance it will ring under the steel mallet as if the tooth were resting on an anvil; 21 322 THE PERIDENTAL MEMBRANE ill {uiotlier case it feels as if the tooth were resting on a cushion. In the first case all of the force of the blow is expended in the condensation of the gold. In the second, a large proportion is lost in the movement of the tooth. If the membrane is thin and the cementum and bone are interlocked, the tooth is firmly supported. If the membrane is thick and the fibers long, as in the first illustration, the blow is dissipated in the sag of the fibers. The tooth is jumping up and down in its socket. The force used is dissipated, the gold is not condensed, and in a very short time an acute inflammation is set up and the tooth becomes very sore to the blows. This the author believes is the explanation of the idea that gold will not preserve teeth for young children. It has often been said that children's teeth are too soft for gold fillings. The difficulty is not with the enamel and dentine, but because of the thickness of their membranes. The gold is not sufficiently condensed to exclude moisture, and the fillings fail. Serious damage also may be done to the membrane. The Museum of the North- western University Dental School contains an object lesson on this point. It consists in a bicuspid with a beautifully condensed and finished gold filling, in a mesial occlusal cavity. The history accompanying it is somewhat as follows: The operation was undertaken for a patient aged about fourteen years. The tooth became exceedingly sore under the mallet; the filling was, however, completed and polished, but a few days later the tooth was picked out with the fingers. The peridental membrane had been literally hammered to death. Stated in scientific terms, the fibers had sawed upon the bloodvessels, exciting an acute inflammation, resulting in complete stasis and the death of the tissue. In all opera- tions where gold is to be condensed in teeth with thick membranes they must be firmly supported so as to be held rigidly against the blow. CHAPTER XXIV THE IMOUTH CAVITY Mucous Membrane. — The mucous membrane lining the mxouth cavity is composed of a layer of stratified squamous epithelium supported upon a tunica propria, which is usually described as composed of two parts^ — the papillary layer and the reticular layer. The epithelium and the tunica propria make up the mucous membrane proper, which is sup- ported upon a submucous layer composed of a coarse network of white and elastic fibers, containing the larger bloodvessels. The Epithelium. — The stratified squamous epithelium is provided with a horny or corneous layer only in the por- tions covering the alveolar process and the hard palate, or, in other words, where the submucosa is firmly attached to the periosteum (Fig. 249). In these positions the horny layer consists of dead cells which have lost their nuclei and whose cytoplasm has been converted into keratin or horny material. These scale-like cell remains are closely packed into a protective layer. There is no distinct stratum lucidum separating the dead from the living cells, as there is in the skin. In the deeper portions the cells possess oval or rounded nuclei and become larger and more polyhedral as the base- ment membrane is approached. The cells of the deepest layer next to the basement membrane are tall and approach the columnar form, but are never much greater in height than width. The deep layer is often called stratum ]\Ial- pighii. The epithelium lining the gingival space and that coverings unattached portions is without the horny laj'er, and the cells are larger and more loosely placed. The poly- hedral cells in the middle portion of the layer show distinct 324 THE MOUTH CAVITY intercellular spaces across which the cytoplasm extends in intercellular bridges. Isolated cells from this region show the broken bridges projecting from their surface, and for this reason have been called "pickle or prickle cells." In these positions the thick- ness of the epithelial layer is usually greater than in the attached portions of the membrane (Fig. 250). Fig. 249 Stratified squamous epithelium covering the alveolar process: C, corneous layer; P, papilla of connective tissue. (About 400 X) Tunica Propria. — The connective-tissue layer of the mucous membrane interlocks with the epithelial layer by means of the tunica papillaris, which is composed of very delicate white and elastic connective-tissue fibers. They are usually about half as tall as the thickness of the epithelium, and about one-third as wide as they are tall. The height and character of the papillae varies greatly, however, in different positions. In the red border of the lip and in the epithelium GLANDS OF THE SUB MUCOSA 325 lining the gingival space they are very tall and narrow, and approach very close to the surface of the epithelium. Over the gums and the palate they are much shorter and wider and do not extend more than half-way through the epithelium. These papilla? contain loops of capillary bloodvessels, and in some special nerve endings are found. Fig. 250 Stratified squamous epithelium from unattached mucous membrane of the mouth. The corneous layer is absent. (About 200 X) Reticular Layer. — The reticular layer joins the papillary layer without any line of demarcation, and is composed of the same kind of tissue, the fibers being arranged in a deli- cate network. Everywhere in the tunica propria are found ducts from mucous glands which lie in the deeper layers. Submucosa. — The submucosa is composed of firm connec- tive tissue in which the white fibers are in large, strong bundles, and elastic fibers are scarce. It contains two plexuses of bloodvessels, both more or less parallel with the surface. The outer is composed of small vessels forming a smalk meshed network, the deeper of large vessels more \\idely separated. Lymphatic vessels everywhere follow the course of the bloodvessels. 326 THE MOUTH CAVITY Glands of the Submucosa. — The siibmiicosa contains a great many small tubular glands. These are distributed widely over the tongue and membrane of the cheek and lip (Fig. 251). They are branched tubular glands, sometimes simple Fig. 251 k. Mucous (jkmd' Epitlielinm [ of mucous. -.L memhrane Blood- vessplfi Epithelium of mucous membrane \/ Cross sections^ of muscle'-~-\{-~. P- '^O. Ffnir ■ ]\ ' foJIicles -yr—Fnt -f-'Epidermis f ^,(ll (1)1(1 s _ _ - - - ^ of muscle ibres Mucous membrane tvith high'' papillse " Longitudinal sections Place irhere stratum corneum begins Section through the upper lip of a two-and-a-half-j^ear-old child. (14 X) THE TONGUE 327 and sometimes compound. The body of the gland is always in the submucosa, though it may extend into the underlying muscle. Some are serous and others mucous, while many of the larger ones contain cells of both types. The secretion of these glands is probably much more important than has been supposed. Nerve Endings in the Mucosa. — Sensory nerve endings of two kinds are found in the mucous membrane. Krause's end bulbs are found in many of the papillae, and other nerves terminate in free endings lying between the epithelial cells. Fig. 252 A section from the side of the tongue: E, epithelium; Sin, submucosa; Bv, blood- vessels; Af, muscle fibers; G, mucous glands. The Tongue. — The tongue is composed of a mass of volun- tary muscle fibers arranged in complicated interlacing bun- dles, covered by the mucous membrane. The most striking characteristics of the mucous membrane of the tongue (Fig. 252) are: (1) The thinness of the submucosa, which holds 328 THE MOUTH CAVITY it closely to the mass of muscle and allows very little move- ment of it; (2) the submiicosa in the dorsal surface contains no glands, though there are glands among the muscle fibers whose ducts pass through the submucosal (3) the presence of the epithelial papilla upon its dorsal surface. The tongue is imperfectly divided vertically on the median line by a band of connective tissue forming the median raphe or septum, which causes the depression at the central line of the dorsal surface. The Muscles. — The muscles of the tongue include two groups — the extrinsic and the intrinsic. The extrinsic mus- cles comprise the genioglossus, the hyoglossus, the styloglos- sus, and the palatoglossus. These are all paired and extend from the skull or the hyoid bone into the tongue. The intrinsic muscles comprise the principal muscles of the tongue, the lingualis. A transverse section through the body of the tongue in the central portion shows a compli- cated network of muscle fibers running in three directions — longitudinally, transversely, and vertically. The longitu- dinal fibers are arranged around the outer portion, forming a cortical layer about 5 mm. thick. These constitute the chief bulk of the lingualis, supplemented by fibers from the styloglossus. The vertical fibers are mostly deeply placed in the central portion on either side of the raphe. They are chiefly derived from the genioglossus and radiate toward the dorsal surface. The transverse fibers are entirely from the lingualis except for a few from the palatoglossus. They arise from the septum and interlace with the longitudinal and vertical fibers. They break up into strands running between the longitudinal fibers of the cortical portion, and spread out to a submucous insertion. The complicated movements of the tongue are accom- plished by the contractions of these sets of muscles. When the longitudinal fibers are relaxed and the transverse fibers contracted the tongue is rolled and extended. When the transverse fibers are relaxed and the vertical fibers contracted the tongue is flattened. The division of the tongue on the median line by the septum allows each half to work inde- THE PAPILLA 329 pendently, so that when the longitudinal fibers are con- tracted on one side and relaxed on the other the tip of the tongue is moved sideways. The Papillae. — The roughness of the dorsal surface of the tongue is caused by projections of the epithelium resting upon the tunica propria, forming the papillae of the tongue. These projections are not to be confused with the connective- FiG. 253 Mucous membrane from the dorsal surface of the tongue of a kitten, showing filiform and fungiform papillse. tissue papillae in the tunica propria of the mucous membrane. They are of three kinds — the filiform and fungiform papillae, which are found over the entire dorsal surface, and the circumvallate papillae, which are limited in number and confined to the posterior portion. The filiform are much the more numerous, especially near the tip of the tongue. They are from 0.5 to 2.5 mm. in height, and often end in brush-like strands of epithelial cells. 3:^0 THE MOUTH CAVITY The fungiform papillse form the red points on the surface of the tongue, especially near the edges, because of the Fig 254 Mucous membrane from the tongue of a rabbit, showing circum vallate papills with taste buds on their sides. Fig. 255 P h s A section of a taste bud: p, pore; g, gustatory cells; ep, epithelial cells; s, sustentacular cells; h, bristles of the gustatory cells. (Schaefer.) THE TONSIL 331 thinness of their epithelium. They are low and rounded in form, from 0.5 to 1.5 mm. in height, and are named from their mushroom-like appearance. Fig. 253, a section from the tongue of a kitten, shows the form of both of these papillae. The circumvallate papillae usuall}^ number nine or ten, and are arranged in a V-shaped form near the base of the tongue, with the apex extending backward. They are from 1 to 1.5 mm. in height and from 2 to 3.5 mm. in width. They are surrounded by a depression, so that the upper surface of the papillae is not much above the general level of the membrane. The Taste Buds.— These are found chiefly on the sides of the circumvallate papillae (Fig. 254), though they are occasionally found in the epithelium of the fungiform papillae and the soft palate, and on the posterior surface of the epiglottis. They are always entirely embedded in the epithelium and extend through its entire thickness. The structures are ovoid in form, with the rounded end toward the connective tissue and the pointed end at the surface, where a small opening, the taste pore, communicates with the mouth cavity (Fig. 255). INIost of the cells are elongated and spindle-shaped, and arranged like the leaves of an onion. Four varieties may be recog- nized. The outer sustentacular cells form the outer layer and are in contact with the epithelial cells. They are elongated, with an oval nucleus near the centre. The inner sustentacular are rod-shaped cells, more slender in form, with a nucleus at the base. The neuro-epithelial cells are elongated, spindle-shaped cells at the centre of the taste bud. The nucleus is at the base of the cell, and from the opposite end a stiff bristle-like process extends through the taste pore. The basal cells are irregular in form with large oval nuclei; they communicate with each other and the sustentacular cells by cytoplasmic bridges. They form the base of the taste bud. The function of the taste buds is probably related to the function of deglutition rather than the sensa- tion of taste. The Tonsil. — In the posterior part of the tongue and the wall of the pharynx is found adenoid tissue in the form of 332 THE MOUTH CAVITY solitary follicles lying in the tunica propria and invading the epithelium. This adenoid tissue forms an organ which Waldeyer has called the lymphatic pharyngeal ring. This tissue is divided into three main masses — that lying in the base of the tongue forming the lingual tonsil, that associated with the palate and lying between the pillars of the pharynx and forming the palatine tonsil, and that situated in the pharynx or pharyngeal tonsil. .^- .^, Epithe- lium TanicK propria Lympli nodule Oblique section "j"'".- .^c;i^. of duct — p--^ - -' of raucous gland Muscle fibres cut — ii^. trans- versely \^ar-iCC>-iCQi-iCC!.-i'H.. : vr-f^T^^ .J ^ ■- */..:■ <^:.rr<«>t-_s»»n:*>*-^;r«K!-?»j»!«i»«»E«WJ8r«cra^ Ift. .. ^1- -^?.^* The lingual plate from Fig. 293. RELATIOX OF THE TEETH TO THE BOXE 189 be studied in this connection, remembering that the bone has been formed and shaped by formation of subperiosteal bone on its surface and subperidental bone at the border of the process and their transformation into Haversian system and cancellous bone. Fig. 293 is cut transversely. Notice that the gingival section has been turned over in mounting. Observe the cribriform plates forming the walls of the alveoli, and the way these are braced against each other and the cortical Fig. 299 The bone between the alveoli cf the mesial and distal roots of the 6rst molar, from Fig. 293. plates by bands of cancellous bone. In accordance with the principles noted, the buccal plate is thin and very compact, while the hngual plate is much thicker, but more open in structure, and the direction of growth has been toward the buccal as the arch of the jaw increased in size. Fig. 297 shows the buccal plate with higher magnifications. Fig. 298 the lingual plate, and Fig. 299 the bone separating the alveoli for the mesial and distal roots of the molar. The third figure of this series shows only the tip of the distal root of the 390 THE TEETH AND DEVELOPMENT OF THE FACE molar, but the arrangement of plates of cancellous bone between the cortical plates is nicely shown. The Maxilla. — In the maxilla the arrangement is exactly on the same plan, the details being different because of the difference in the shape of the bone. THE GROWTH OF THE JAWS It has long been noted that at birth the mandible is straight, and with the eruption of the teeth the ramus develops and the body increases in size. In this process the thickness of the bone is increased from the mental foramina to the alveolar border, and the body of the bone approaches a right angle with the ramus. When the teeth are lost or lose their function the alveolar process is destroyed and the bone reduced in thickness from above downward until the mental foramen comes to lie on the upper surface of the bone. The mandible performs two functions, a respirator}^ and a masticatory function, and it should be remembered that these are influential in its development. The object of this section is to give some conception of the direction of growth in the development of the bones of the face and the way in which the changes are brought about. This can best be done by studying the series of skulls from childhood to old age, in which the outer cortical plate has been removed so as to show the developing teeth in their crypts and the relation of the forming teeth to those already in occlusion (Figs. 300 to 314). At birth all of the teeth except the second and third molars have begun to develop, and their tooth germs are lying embedded in the cancellous substance of the maxilla. In the upper jaw they occupy almost all of the space to the floor of the nose and orbit, and there is little if any indication of the maxillary sinus (Fig. 300). Each tooth germ is enclosed in a separate crypt, the wall of which is formed by a cribriform plate. The walls of the crypts are braced against each other and the cortical plates of the maxillae by spicules of cancellous THE GROWTH OF THE JAWS Fig. 300 391 MaxilliB at about eight inonlhs after birth, showing the unerupted tooth. 392 THE TEETH AND DEVELOPMENT OF THE FACE hone surrounding medullary spaces. As the tooth develops within its crypt, pressure is exerted and the crypt wall is pushed backward through the cancellous bone. Growth Force.^ — The force exerted by the growing tooth is the result of the multiplication of cells in the tooth germ, and is exactly comparable to the forces exerted by multi- plication of cells in any position. For instance, the force exerted by the multiplication of the cells in the rootlet of a Fig. 302 Maxillae at about one year. plant is sufficient to force pebbles aside and make an opening through hard packed earth. Some attempts have been made to measure the amount of force, but we can only say that it appears to be considerable, acting through short range. How this force is generated has been a matter of much speculation and investigation It shows some points of similarity with the swelling of wood fibers when water is added. It apparently is related to osmosis, and has some THE GROWTH OF THE JAWS 393 direct relations to blood pressure. It is certainly a very complicated matter, with chemical affinities at the bottom of it. Forces Influencing Bone Growth. — While the growing tooth germs are producing force which causes conditions of stress of the cortical plates, the growth of the tissues within the mouth — the tongue and the associated organs — is exerting Fig. .-303 Maxillae at one and one-half years. pressure upon the lingual surfaces of the bone. The muscles attached to their surfaces transmit force to the bone through the periosteum, and the functions of mastication, deglu- tition, and respiration are acting upon them. All of these are mechanical stimuli, to which the connective-tissue cells respond. In all the process of development the growth is the result of all the forces to which the bones are subjected, perfectly distributed through the substance of the bone bv 394 THE TEETH AND DEVELOPMENT OF THE FACE the agency of normal occlusion. Any lack of harmony in the proportion of these forces may allow the teeth to meet, when they erupt, outside of the normal influence of their cusps, causing the beginning of malocclusion. Any mal- occlusion disturbs the balance in the distribution of forces, Fia. 304 Maxillae in the second year, showing the relation of the erupting teeth. Note the relation of the crypt of the second molar to the inferior dental canal. and results in a disturbance of the development of bone, which progresses during the entire period of development. This must result in the lack of balance in the proportions of the features which will be proportionate to the mal- occlusion. THE GROWTH OF THE JAWS 395 It has been natural and almost inevitable, because of their hardness, to think of bones as sohd and unchanging. In the study of these skulls the bones of the face must be viewed not as solid and rigid, but as containing millions of active cells which are continually building and rebuilding their substance. Fig. 305 The complete temporary dentition (about three years), showing the relation of the developing permanent teeth. Usually somewhere between the seventh and ninth months after birth the growth of the central incisors causes the absorp- tion of the roof of their crypts, and the tooth moves occlusally, cutting through the soft tissues (Fig. 301). The formation of cementum on the surface of the root and of bone on the wall of the crypt attach the connective-tissue fibers and form the beginnhig of the peridental membrane. As the tooth moves occlusally the bone grows up around it from the circumference 396 THE TEETH AND DEVELOPMENT OF THE FACE of the crypt wall, converting it into the wall of the alveolus. The root is not fully formed and the conical pulp filling the funnel-like end exerts force by the multiplication of cells and the blood pressure, which cause the tooth to move occlusally and the bone to grow in that direction. At the same time the pressure of tongue and lips exerts pressure on Fig. 306 The complete temporary denlition and the first permanent molar. Note the relation of the bicuspids to the temporary molars. (In the seventh year.) the surfaces of the tooth and bone, influencing the direction of bone growth. The jaw increases in thickness in the occlusal direction and grows forward and outward. At the same time the growth of each successively distal tooth is exerting pressure upon those already erupted, causing them to move farther in the occlusal direction. In Figs. 303 and 304 notice the way in which the crypt walls are pushed downward by the THE GROWTH OF THE JA^yS 397 development of the tooth root until the inferior dental nerve lies between the floor of the crypt and the cortical plate of the lower border. In this way enough pressure may be produced Fig. 307 Front view of the skull shown in Fig. 303. Note the relation of the permanent incisors and cuspids to each other and the roots of the temporary teeth. 398 THE TEETH AND DEVELOPMENT OF THE FACE to cause reflex nervous symptoms, which commonly precede the eruption of the temporary molars, and so development continues until all of the temporary teeth are in place. About Fig. 308 Dentition in the eighth year. Note the position of the cuspids and compare with Fig. 310. THE GROWTH OF THE JAWS 399 the sixth year the first permanent molars take their place at the distal of the temporary teeth and their cusps interlock (Fig. 305). The importance of these teeth can scarcely be Fig. 309 The left side of the skull, shown io Fig. 30S 400 THE TEETH AND DEVELOPMENT OF THE FACE overstated. They are not only to be the chief means of mastication during the period in which the temporary teeth are lost and replaced by their successors, but they are to Fig. 310 Dentition in the eleventh year. Note the growth of the cuspids and bicuspids. The second molar is about to erupt. THE GROWTH OF THE JAWS 401 maintain the relation of the jaws to each other. The way in which these teeth lock determines the balance between the forces exerted by the action of the muscles attached in the region of the ramus, and those in the region of the svmphvsis (Fig. 306). Fig. 311 Dentition in the thirteenth year. Note the relation of the bicuspid crown to the roots of the lower temporary molar. A deviation from the normal relation of these teeth will entirely change the direction of the forces, and will be mani- fested by a modification in the development in the bone. In the skull at this period the bicuspids are seen lying below the temporary molars, and the second molar developing at 26 402 THE TEETH AND DEVELOPMENT OF THE FACE the distal of the first. Their growth is transmitted through the teeth to the alveolar process, and the addition of bone results. The same skull viewed from in front (Fig. 307) shows Fig. 312 The dentition of a young adult. The third molars have not erupted. (About fifteen years.) THE GROWTH OF THE JAWS 403 the relation of the permanent incisors and cuspids to the tem- porary ones. In the lower jaw the temporary centrals have been lost and the permanent ones are forcing their way between the temporary laterals. The crowns of the centrals Fig. 313 Adult dfentition. Note the distance from the apices of the incisors to the lower border of the mandible and the floor of the nose. are wider than those of the teeth that were lost, and they consequently exert pressure upon the mesial surfaces of the laterals, pushing them apart and carrying them upward and forward. Studv the relation of the lower centrals, laterals, and 404 THE TEETH AND DEVELOPMENT OF THE FACE cuspids in the development of the arches at from six to ten years. Notice that the roots of the central are not fully formed, that the lateral lies to the lingual of the temporary lateral root, and with its mesio-occlusal angle below the distal surface of the central. The development of the cuspid has pushed the crypt floor through the cancellous bone until it has reached the solid cortical plate, and still the for- mation of the crown is not quite completed. The six teeth Fig. 314 Edentulous jaws, showing loss of alveolar process. form a triangle of which the centrals are the apex, and the cortical plates from cuspid to cuspid the base. The com- pletion of the roots of these teeth will carry the temporary teeth, alveolar process and all, upward, forward, and out- ward, thus increasing the distance from the mental foramen to the symphysis and enlarging the arc of the law from cuspid to cuspid. In the same skull notice the relation of the upper incisors and cuspids to the corresponding temporary teeth. They lie THE GROWTH OF THE JAWS 405 to the lingual of the roots of the temporary teeth, the lateral a little to the lingual of the central and cuspid. The cuspid has pushed back the floor of its crypt until it is braced against the solid bone at the base of the malar process. The growth of these teeth will first cause the temporary teeth to move occlusally, the bone growing from the border of the process to follow them. In this growth the distance from cuspid to cuspid is increased and spaces appear between the tempo- rary incisors some time before they are lost. If such spaces do not appear, the development is not progressing normally, and artificial force should be applied to stimulate bone growth. If this is not done the permanent teeth are sure to come in more or less rotated and out of position. In Figs, 308 and 309 the incisors have been pushed ofi' and the permanent ones are beginning to move occlusally. Notice the relation of the floor of the crypt to the floor of the nose, and the root has scarcely begun to develop. In the adult skull (Fig. 302) there is almost as much space from the apex of the root to the floor of the nose as there is now from the border of the alveolar process to the floor of the nose. The result of the growth of the cuspids' roots is shown by comparing Figs. 307 and 308 with Fig. 310. The Importance of Proximal Contact. — The proper contact of the teeth upon their proximal surfaces is necessary for this development. If, for instance, the mesial angle of the lower lateral fails to engage with the distal surface of the central, but slips by to the lingual, the growth of the cuspid will push it farther and farther past the central instead of enlarging the arch. One of the cogs in the mechanism has slipped, and the growth of bone cannot later be expected to make room for the crowded teeth. In the next stage of groT\i:h the increase in size is from the mental foramen to the ramus, and is largely influenced by the development of the roots of the bicuspids and the second molars. Figs. 309 and 310 show the relation of the second molar to the distal surface of the flrst, and it will be seen that its growth exerts force upon the first 406 THE TEETH AND DEVELOPMENT OF THE FACE Fig. 315 Fig. 316- Figs. 315 and 316 were photogiaphed in the same relative size, to show the amount and direction of growth, with the development of the full permanent dentition. THE GROWTH OF THE JAWS FiQ. 317 407 Fig. 318 Figs. 317 and 31S -were pliotographet: in tlio same relative size, to shew the amount and direction of growth, with the deveiupmeut of the full permanent dentition. I'la. 319 Fig. 320 Figs. 319 and 320 were photographed in the same relative size, to show the amount and direction of growth, with the development of the full permanent dentition. Fig. 321 Fig. 322 Figs 321 and 322 were photographed in the same relative size, to show the amount and direction of growth, with the development of the full permanent dentition. 410 THE TEETH AND DEVELOPMENT OF THE FACE molar, and this is transmitted through the arch by means of proximal contact. Notice the inclination of the bicuspid roots, which help to carry the growth in the same direction. After the second molar is in place the growth of the third should exert the same force and room be provided for it (Fig. Fig 323.— Two years. Fig. 324. — Three years. Fig. 325.— Six years. Fig. 326.— Ten years. Maxillae photographed from the median line in the same relative size, to show the amount and direction of growth. THE GROWTH OF THE JAWS 411 Fig. 327.— Twelve years. Fig. 328.— Adult. Maxillae photographed from the median line in the same relative size, to show the amount and direction of growth. 311). The muscular action of the lips and tongue are specially important in these last stages of growth, and particularly the forces that are generated by the action of the muscles in respiration and deglutition. The activity of the connective- tissue cells in the bone require mechanical stimuli for their maintenance, and as the muscular action is vigorous or deficient, the growth of bone mil be full and normal or imperfect and unbalanced. It appears often that the bone activity becomes so sluggish that the growth of the third molar cannot produce the effect it should, and it remains impacted. A comparison of figures will show that while room has been made for the third molar, all of the upper teeth have moved downward, forward, and outward, and the lower ones upward, forward, and outward. Compare the distance from the apex of the incisor roots to the floor of the nose and the lower border of the mandible in Figs. 312 and 313. This process may be more fully realized by comparing 412 THE TEETH AND DEVELOPMENT OF THE FACE the front views of the skulls (Figs. 315 to 322). They were all photographed with the same lens and bellows length, so as to make the pictures of the same relative size as the skulls. Notice the increase in distance from the floor of the nose and the floor of the orbit to the edges of the upper Fig. 329 Bone from the buccal plate of the mandible of a young sheep, showing transforma- tions of bone: 1, subperiosteal bone; 2, Haversian system bone; 3, Haversian system bone, becoming cancellous. incisors, and from the lower border of the mandible to the edge of the lower incisors. It will be seen that if the infant mandible were placed in relation to the adult mandible it would lie entirely within the arch and in the mouth cavity, while in the upper the temporary incisors in Fig. 322 would be some place in the nasal cavity. In all of this growth the THE GROWTH OF THE JAWS 413 size of the air spaces increases with the movements of the teeth, the floor of the nose and palate growing downward and developing. This may be shown in Figs. 323 to 328, in Fig. 330 The record in the arrangement of the lamellae of the growth of the mandibles. A decalcified section from near the lower border of a human mandible. which the right half of the maxilla has been removed from dissected skulls and photographed from the median line. Tissue Changes in the Physiologic Movements of the Teeth. — All that has been said in regard to bone growth must be 414 THE TEETH AND DEVELOPMENT OF THE FACE recalled in order to obtain a conception of the manner in which these movements of the teeth and the development of the bone are accomplished. Bone laid down under FiQ. 331 A decalcified section from the lingual vertical plate of a human mandible, showing the arrangement of lamella as a record of growth. the periosteum and the peridental membrane has been transformed into Haversian system bone and then made cancellous, as illustrated in Fig. 329, which is taken from the buccal plate of the mandible of a young sheep. THE GROWTH OF THE JAWS 415 Reversed changes have also been going on, the periosteum cutting into the Haversian bone by absorption and the cancellous bone being condensed into Haversian system bone. These changes leave a record in the arrangement of Fro. 332 Cancellous bone from a decalcified section of a human mandible, showing reconstructions to change the direction of the spicules. the lamellse, and may be studied in decalcified sections (Figs. 330 to 333). Even the direction of the spicules of cancellous bone are being constantly changed by absorptions and rebuilding to adjust them to changes of stress. 416 THE TEETH AND DEVELOPMENT OF THE FACE While the temporary teeth are moving occlusally, bone is laid down under the peridental membrane at the border of the alveolar process, which is at once cut out by absorptions and replaced by Haversian system bone (Fig. 217). The alveolar process becomes a veritable patchwork, as shown in Figs. 234 and 335. The permanent tooth developing in its crypt produces conditions of pressure, and osteoclasts appear in Fici. 3;33 Decalcified section of cancellous bone from a human mandible, showing absorptions and rebuildings, changing the direction of the spicules. all the medullary spaces, around and above the crypt, and through the alveolar process, as well as on the crypt wall. They are more active in the medullary spaces, cutting away the spicules of bone, thinning and cutting apart the crypt wall, and allowing it to be bent and pushed back. Fig. 336 shows the alveolar process on the lingual side of the temporary incisor, and illustrates the enlargement THE GROWTH OF THE JAWS 417 of the medullary spaces preparatory to the eruption of the permanent tooth. Fig. 337 shows the labial plate of the process, and notice that the bone is being formed under the Fig. 334 A longitudinal section through the tip of the alveolar process of a temporary tooth about ready to be lost: D, dentine; Cm, cementum, showing absorption and rebuild- ing; Pd, peridental membrane; B, bone growing occlusally at the border of the process; Hb, rebuilt Haversian svstem bone. 27 FiQ 335 A longitudinal section through the temporary alveolar process, which is growing occlusally to follow the temporary tooth. It is from the same series as Fig. 334, but shows more of the bone. Study the absorptions and rebuildings, as shown in the arrange- ment and character of the lamellae. Pd, peridental membrane: Po, periosteum. THE GROWTH OF THE JAWS 419 420 THE TEETH AND DEVELOPMENT OF THE FACE THE GROWTH OF THE JAWS 421 422 THE TEETH AND DEVELOPMENT OF THE FACE periosteum and at spots under the dental membrane, while the substance of the bone is being destroyed. When the tooth is finally pushed off from the gum all but a few bits of the alveolar process have been destroyed, and as the permanent tooth comes through, bone formation begins at the border, patching on to the remains of the old process (Fig. 338). In studying the absorption of bone around the crypt walls, it has been noted that the osteoclasts appear first in the cancellous bone (Figs. 218 and 219), surrounding the crypts and outside of it. Absorptions here remove the spicules which brace the crypt wall, and cut through the wall in such a way as to allow it to be pushed back through the weakened substance. In the same way in the movements of the teeth, absorptions appear first in the spaces outside of the cribri- form plates of the alveoli, until the remaining bone is weak- ened sufficiently to spring under the pressure. All of the sections of the mandible should be studied as a record of these bone transformations, and especially in orthodontia it should be remembered that appliances are used not to push the teeth through the bone as a post would be pushed through the mud, but to supply mechanical stimuli to living cells whose activity will result in bone growth, carrying the teeth into their proper positions, and finally that teeth will remain only in the position in which all of the forces to which it is subjected are balanced. PAET II DIRECTIONS FOR LABORATORY WORK (TWENTY-FIVE PERIODS IN THE LABORATORY) INTRODUCTION It is assumed in this work that the student has had a course in general histology, including laboratory work, that he is familiar with the technique of handhng the microscope, the technique of staining and mounting sections, and that he is able to recognize at once the elementary tissues. The same outfit is required as for general histology, including slides and blank labels for them; cover-glasses; teasing needles; forceps; section lifter; a tube of balsam; a funnel; pipette; filter paper and lens paper; 6 one-ounce reagent bottles containing xylol, absolute, 95, and 70 per cent, alco- hols, hematoxylin, and eosin; at least two chip butter dishes that can be used for staining; a box for the slides; a notebook; a hard and a soft drawing pencil; a good eraser; and a piece of clean, soft linen for wiping slides and cover- glasses. Teeth for Grinding. — It is difficult to obtain satisfactory teeth for the grinding of microscopic sections, and the student should bring to the laboratory a number of suitable teeth from which selection can be made. Old, dry teeth are absolutely useless for the purpose, however perfect their structure may have been. When a tooth has been extracted for some time the tissues dry out, giving up a considerable amount of water, and consequently shrink. The shrinkage of dentine and enamel is unequal, and the result is a cracking 424 DIRECTIONS FOR LABORATORY WORK of the tissue. The observation of the teeth in any skull will reveal cracks in the enamel that may be seen with the naked eye, the tooth often splitting lengthwise. Besides the cracks that can be seen, the tissue is full of microscopic cracks. When the grinding of sections from such teeth is attempted, before the section is reduced to sufficient thinness for micro- scopic observation, the enamel will break to pieces and be lost. A tooth that is to be used for grinding must be placed in solution as soon as it is extracted, and never at any stage of the process be allowed to dry, until ready for mounting. Any solution that will prevent decomposition will do for this purpose. The best that I have found is a 4 per cent, formaldehyde in 50 per cent, alcohol. This may be roughly prepared by diluting 95 per cent, alcohol with an equal volume of water and adding one part of formalin to nine parts of the diluted alcohol : Alcohol 45 c.c. Water 45 c.c. Formalin 9 c.c. This solution not only prevents the drying, but has a hardening action on the organic matter, which facilitates the grinding. Teeth may be preserved in this indefinitely. Teeth Required. — From his collection the student should select for grinding an incisor or cuspid, a bicuspid, and a molar. The teeth should be free from caries and their crowns as perfect as possible. The Relation of the Section to the Crown. — The practical value of the study of ground sections depends upon obtaining from them a knowledge of enamel rod directions in relation to the tooth crown as well as the section. In operating the teeth are looked at from their outside surface, but the operator needs to see in the enamel not simply a hard and extremely dense tissue, but a tissue made up of minute rods whose general direction he knows beforehand. If a tooth is selected and a section cut from it in a known position, and the relation of the section to the crown remembered, the direction of enamel rods can be placed in relation to the PREPARATION OF GROUND SECTIONS OF TEETH 425 entire crown as well as to the section. This is one of the objects to be sought in the making of the outline drawings. Location of the Section. — Having selected the teeth for grinding, the next step is to locate the position and direction of the section. This must be so placed as to cut the enamel rods in their length. The section from the incisor or cuspid should be ground labiolingaally, but the section from the molar and bicuspid may be ground either buccolingually, mesiodistally, or diagonally. The surface of the tooth should be considered, and the section placed in an area in which the student desires to discover the enamel rod direc- tions and the structure of the tissue. The line of the section should now be marked on the tooth with India ink and a fine pen. The Drawings of the Teeth. — After marking the position of the section the tooth should be carefully and accurately drawn, showing the position of the section as seen from the axial and occlusal surfaces. Grinding of the Section. — Every institution should have a machine for the preparation of ground sections, but such a machine is too delicate an instrument to be handled by students. In the appendix will be found a chapter written by Dr. Black describing the grinding machine and the technique of its use. If one is available, the student may have his sections ground for him and returned ready to mount, or he may grind them himself, using the following technique: Preparation of Ground Sections of the Teeth.— For this work the student should have two large corundum stones not less than four inches in diameter, one of "C" and one of "E" grit. Corundum is very much better than carborundum for this purpose. In grinding the stone should be kept revolving slowly and moistened with a stream of water. Holding the tooth against the flat side of the coarse stone with the fingers, the tissues should be rapidly ground away until the position marked for the section is reached, when the fine stone should be substituted and the grinding continued just enough to remove the scratches. The surface should now be polished on the Arkansas stone until a very perfect surface 426 DIRECTIONS FOR LABORATORY WORK has been obtained. Wash the specimen clean and immerse in several changes of 95 per cent, alcohol, and leave in absolute alcohol in a closed bottle for several hours or over night. Harden a drop of balsam on the centre of a clean slide by warming it over a Bunsen burner to evaporate the xylol. When the slide is cool the balsam should be neither sticky nor brittle. Now remove the tooth from the alcohol, wipe it dry, and, placing it on the balsam with the polished surface next to the glass, gently warm the slide until the balsam is thoroughly softened, and press the tooth down against the glass and clamp it firmly in position, using a spring clip. Set it away to harden thoroughly, when the grinding may be continued. Holding the slide parallel with the surface of the coarse stone, the tissues may be rapidly removed until the section is about as thin as a calling card, when the fine stone should be substituted and the section reduced to the required thin- ness. It should not be more than twenty microns in thick- ness. In the final stages progress of the grinding may be followed with a hand magnifying glass. Finally the surface should be polished on an Arkansas stone. The specimen should now be washed with alcohol, the balsam removed with xylol, and brought to the laboratory in 95 per cent, alcohol, where it is to be etched and mounted according to the directions. Every step in the above technique is important and must be followed with minute care and accuracy. Not least important is the cleaning of the slide. It sometimes happens that the section will be loosened from the glass before the grinding is completed. This is usually due to some fault in the technique. When it happens it is best to finish the grinding without attempting to refasten the section to the slide. To do this the section should be held against the flat side of the stone, using a fine-grained cork, a piece of box- wood, or some similar material. The danger of breaking the section, however, is much greater. The Preparation of Transverse Sections of the Root. — For this purpose one of the flattened roots furnishes the best DRAWINGS 427 material, as, for instance, the mesial root of a lower molar, the root of a lower bicuspid, or of an upper second bicuspid. Holding the root in a vice by the remains of the crown, with a metal saw, saw off the tip of the root, removing an eighth of an inch or less. Then saw off as thin a slice as possible. In the same way saw^ out at least two other sections, one from the gingival and one from the middle third of the root. These should be dropped into a bottle of formalin-alcohol until the grinding is completed. The grinding is easily accomplished on the flat side of the corun- dum stone, holding the section on the finger or under a cork. The last grinding should be done on the fine Arkansas stone. Transverse sections of the root are easily ground and can be made very thin. Manner of Working in the Laboratory. — In no place in the world can time be wasted more easily than in the histological laboratory. The student should take the attitude of an original investigator and study out the material for himself as far as possible, remembering that he has a far better opportunity than the man who worked out the details of these structures. He must constantly try to picture the structure, and imagine how it would appear if sectioned in another direction. Drawings. — Drawings from the microscope are made not simply to occupy the student's time, nor as a record of what he has done, but to make observation more accurate and detailed, and to fix the impressions of structure more per- fectly in mind. Many students excuse themselves for careless and slovenly work by saying that they are not artists. Anyone without any knowledge of the principles of art can in a very short time acquire the ability to make excellent microscopic drawings. A few principles of pro- cedure will help greatly. The first of all is that a light line can always be made darker, therefore the drawing should always be kept light until the later stages. After selecting a field, draw lightly the outline of the principal masses and then the outKnes of the smaller ones. 428 DIRECTIONS FOR LABORATORY WORK In this way the proportion of objects in the field and their relation to each other can be maintained. Never draw any detail such as individual cells, nuclei, etc., until all of the outlines are completed. Then work in the details in the darker colored areas. The making of the outlines is by far the most important stage in the drawings. Each outfit should contain a 6 H and an H-B pencil and a good eraser, which must be kept clean. The pencils should be kept sharp and always used with a light touch upon the paper. The beginner always tends to start his drawing by making a circle. This should be avoided, for it is objects that are being studied, not fields, and in many cases the object cannot be bounded by a circle. There is also a ten- dency to represent the object smaller on the paper than it appears in the field. The prime qualities in a microscopic drawing are accuracy and correctness of detail. The drawings are made to show all the detail of structure that can be observed. It often happens that a drawing that looks very well shows very little knowledge of the structure of the tissue which it represents. Stencilled Laboratory Notes. — In fifteen years of teaching the author has found stencilled notes on the daily work in the laboratory of very great assistance. There are always variations in the appearance of the material which cannot be anticipated before the sections are cut. Very often some- thing will be seen unusually well that would not be men- tioned in the text-book. Difterent stains may have been used which would change the appearance of the tissues, and for all of these things and many others daily notes are very convenient. USE OF DIRECTIONS FOR LABORATORY WORK At the beginning of the laboratory period the first thing to be done is to read through the directions for the day's work. The amount of work for the dav is then clearlv in mind, and ETCHING AND MOUNTING OF GROUND SECTIONS 429 all the steps in any procedure that is to be undertaken are understood at the beginning. It is necessary to divide the time available, so as to accomplish the work indicated for the day. PERIOD I Drawings of Tooth Surfaces Showing the Position of Sections. — The object of these drawings is to show the relation of the section to the crown from which it is ground, so that in studying the enamel rod directions as seen in the sections, they may be referred to the entire crown. The drawings should be made from five to ten times natural size, and must be made accurately to scale (Fig. 339). Measure the length and the breadth of the tooth and lay out a rectangle, say eight times these dimensions, to serve as a guide in drawing. If the tooth is marked for a buccolingual section, stick the apex of the root on a bit of wax and place the tooth on the table with the buccal surface toward you. Do not change its IDOsition until the drawing is completed, for to do so would change lights and shadows. After getting the outline accur- ately, work in the shadows so as to give the drawing round- ness. Remember in doing this that you can always make it darker, but you cannot erase without injuring the neatness of the drawing. When the drawings are completed the sec- tion is ready for grinding, which must be done outside of the laboratorv, following the directions in Introduction to Part II. PERIOD n Etching and Mounting of Ground Sections. ^ — At the desk will be found 1 per cent, hydrochloric acid, dilute ammonia, and vaseline, which are the only reagents not included in the outfit and required for this work. The sections are brought to the laboratory ground and ready to mount. Fill one of the dishes with water and carefully wash the specimen free from all debris of grinding. Dry the section between filter 430 DIRECTIONS FOR LABORATORY WORK Fig. 339 D/ST/IL SURFACE BUCCAL OCCLUSAL SURFACE BUCCAL MARGIN DISTAL M. MESIAL M. LING LINGUAL M. 8 DIAMETERS Drawings of occlusal and axial surfaces of a tooth, to show the relation of the section to the tooth. (Drawn by W .A. Offil, 1910.) ETCHING AND MOUNTING OF GROUND SECTIONS 431 papers, so as to remove all moisture from the surface. Fill one dish with 1 per cent, hydrochloric acid and the other wdth dilute ammonia. Put a very little vaseHne upon the tip of the finger, and holding the section by the root portion, cover one surface of the crown portion with a very thin layer of it. In doing this the vaseline should be wiped from the centre toward the edges of the section, so as to prevent it from running over on to the other surface. The vaseline is to confine the action of the acid to one surface of the enamel. Holding the section by the root portion, immerse the crown in the dilute acid for thirty seconds, or until minute bubbles can be seen forming upon the surface. Remove and immerse at once in the dilute ammonia for a minute. Remove the vaseline by carefully wiping the section vriih absolute alco- hol or ether, and immerse in 95 per cent, alcohol. In this it should remain while the slide and cover-glass are being prepared. Obtain from the desk a cover-glass long enough to cover the entire section and carefully clean both slide and cover-glass. On the centre of the slide place a drop of balsam that is as long as the section. Holding the slide over a Bunsen burner or alcohol flame, warm it gently so as to evaporate the xylol. In this process the drop will spread out over the slide and the direction of spreading may be guided by the heat. Allow the slide to cool and test the hardness of the balsam with a teasing needle or the finger nail. When cold the balsam should be just soft enough to take the imprint of the needle or nail, but not be sticky. If it is sticky it must be reheated; if, on the other hand, it is brittle enough to chip, it must be scraped oft' from the sUde and the process tried again. In the same way prepare a film of balsam on the cover-glass. Remove the section from the 95 per cent, alcohol and dry it for a few minutes in the air (after wiping with filter paper). Place the section, etched side up, upon the balsam on the slide, and place the cover-glass on it balsam side doicn. Warm the slide gently over the flame, while pressing the cover-glass down with the handle of a teasing needle. As the balsam is warmed, the slide and cover-glass are brought together, forcing the balsam 432 DIRECTIONS FOR LABORATORY WORK out to the edge of the cover-glass in all directions. All excessive balsam should be squeezed out at the edges. Place on the cover-glass a small piece of blotting paper or a layer of cork, adjust some sort of a spring clip and put the section away until the balsam is entirely hard. When the balsam is entirely hard the excess may be removed by gently scraping with a knife blade and wiping with xylol. The section should now be labelled with the name of the tooth, the direction and position of the section, the student's name and number, and the date. The mounting in hard balsam greatly improves the value of the section, for the dentinal tubules and the lacunae of the cementum are left filled with air and can be more easily studied. Sections may, however, be mounted in the ordinary way, in soft balsam. If the section is broken or extremely thin, soft balsam should be used. PERIOD m Outline Drawings from Ground Sections. — The object of the outline drawing is the study of the dental tissues, their distribution, portion of the tooth formed by each, their relation to each other, and the coarser points of their struc- ture. To get the value from this work the drawings must be made very accurately to scale and as large as the note book page will allow. With the bole gauge or a millimeter rule measure accurately the length of the section, multiply this by eight or ten, and mark the length on a page of the draw- ing book. Measure the width of the section at the point of the greatest diameter and multiply this by the same factor. Using this for the width and the previous measure- ment for the length, lightly draw a rectangle, which is to be used as a guide in the construction of the dra^\^ng. The success now of the drawing depends on the accuracy and number of the measurements. First measure the vertical distance from the incisal edge to the gingival line on one side of the section, and then on the other, and mark these on the sides of the rectangle. OUTLINE DRAWINGS FROM GROUND SECTIONS 433 This will give the relative length of root and crown and the difference, if any, in the position of the gingival line on the two sides. Measure the vertical distance from the most prominent point on the axial surface to the incisal edge or the tips of the cusps. And so on, making every measurement that can help in the formation of the drawing. In this way the outline of the section should first be traced inside the rectangle, then the dento-enamel junction, then the pulp chamber is shown, and finally the cementum. Before drawing the outline of the cementum, the section should be placed under the microscope, using the low power, and the cementum should be observed studying it from the gingival line on one side of the section to the gingival line on the other. It would be a waste of time to attempt to fill in the structure of the tissue of the entire outline, and ofily certain things are to be shown in these drawings. For that reason fill in three portions of enamel and dentine and three portions of cementum and dentine, using the low power objective. Study first the bands of Retzius (page 60), and lightly indicate their direction. Study the enamel rod direction, beginning at the gingival line at one side and following it around the crown to the other side. In a portion at the incisal edge, or on the occlusal surface, indicate the rod directions, and in the same way show them in a portion near the centre of the axial surface on one side and near the gingival line. Follow the dentinal tubules which end next to the portions of enamel which have been filled in to the point where they open into the pulp chamber, and indicate their direction (page 171). In the same way fill in three portions of the cementum and the dentine under them — one in the gingival line, one near the middle of the root, and one in the region of the apex (Fig. 340). If any portion of the section has been lost in grinding, that portion should be indicated by dotted lines, and in the same way, if a portion of the crown has been lost by wear, the original form may be added in dotted fines. OutUne drawings should be made from each of the three classes of teeth — one from the incisor or cuspid, one from a 28 434 DIRECTIONS FOR LABORATORY WORK bicuspid, and one from a molar, and a laboratory period should be devoted to each drawing. Fig. 340 ENAMEL ---ENAMEL RODS 'M~ DENTINE {TUBULES) -PULP CHAMBER CEMENTUM Outline drawing of longitudinal section, made as a study of the dental tissues. (Drawn by E. J. Schnaidt.) MINUTE STUDY OF ENAMEL AND DENTINE 435 PERIOD IV Isolated Enamel Rods. — Obtain from the desk a fragment of enamel which has been broken in the direction of the rods. Place a drop of distilled water or glycerin on the centre of a clean slide. jNIoisten the broken surface with a drop of water and lightly scrape it with the blade of a broad, sharp, chisel, holding the edge parallel with the surface and the shaft at right angles to it. Dip the edge of the chisel in the drop of liquid on the slide, and the scrapings will be left. Cover with a cover-glass and study with the high power, using a small diaphragm. Fragments of enamel will be found made up of broken rods, some single and others in groups. Note the diameter of the rods and the appearance of the cross-markings, which will be seen if the light is properly adjusted. Draw as seen with the high power. Repeat this operation, using enamel that has been immersed in 1 per cent, hydrochloric acid foi a number of hours. Com- pare the appearance of the rods with those of the former specimen and make a drawing as seen with the high power. Find an old tooth with a large carious cavity, remove the softened dentine without touching the enamel if possible. Lightly scrape the whitened inner surface of the enamel next to the cavity and mount the scrapings as before. Compare the appearance of these rods isolated by the action of caries with those of the previous specimen. Notice that the cross- markings are more distinct and the expansions and con- stiictions of the rods more prominent. Draw a few of the rods as seen ^\'ith the high power, using the small diaphragm . PERIOD V Minute Study of the Enamel and Dentine. — Select a field from one of the ground sections where the specimen is very thin, and, if possible, where the entire thickness of the enamel plate can be seen in one field with the f objective. 436 DIRECTIONS FOR LABORATORY WORK To select this field all of the enamel in the three sections should be carefully studied with the low power, and the one Fia. 341 DENTO-ENAMEL ..^ni\?\ Sr«P>GHT ROI '•;t' pEI'lNPu TL'SULI High-power drawing of the enamel. (Drawn by A. B. Hopper. 1902-03.) MINUTE STUDY OF CEMENTUM AND DENTINE 437 chosen in which the rods can be seen best and can be most easily drawn. Having selected the field, study the enamel with the high power, beginning at the dento-enamel junction. Note the form of the dento-enamel junction and the relation of the two tissues at this point. Note the diameter of the enamel rods and estimate it, using a red blood corpuscle as a standard of measurement. Note the striation of the enamel (page 57). Using both the low and the high power, draw as accurately as possible the enamel from the surface to the dento-enamel junction, showing all the details of structure that can be made out. The drawing should be made as long as the page will allow, and need not be more than an inch wide, and should include just enough of the dentine to show the dento-enamel junction and the character of the dentine at that point (Fig. 341). Notice the diameter of the dentinal tubules, comparing them with the red blood corpuscles and the enamel rods. Note the amount of matrix that separates the tubules. Observe the forking and the anastomosis of the tubules as they approach the enamel, and follow them as far as possible. PERIOD VI Minute Study of the Cementum and Dentine. — With the low power study the cementum in the three specimens, looking for all the details of structure that can be made out (see page ISl). In the gingival portions and often well toward the apex, especially if the tooth is from a young person, the cementum will be very thin and almost structureless in appearance. ^Yith the high power, fine lines parallel with the surface may be seen, which indicate the lamellae. In the apical portion the cementum becomes much thicker, and it will be seen that each layer is thicker and consequently more easily seen. Little black spots looking like spiders will be found in larger or smaller numbers. These are the lacunae \\dth the canaliculi radiating from them. They were filled in life by cement corpuscles. Look for embedded 438 DIRECTIONS FOR LABORATORY WORK fibers of the peridental membrane. In all of this work each field should be studied with both the low and the high power. <*M- A.e Hc\s yr Fig. 342 — CEMENTUM 4 \ ^^<^y^^ 't^ - GRANULAR LAYER OF TOMES DENTINE PULP CHAMBER Cementum and dentine. (Drawn by H. J. Lund and A. E. Hopper.) DRAWINGS OF TYPICAL CAVITY WALLS 439 The inner layer of the cementum next to the dentine is clear and structureless, and the dentine adjoining it appears with the low power as a granular layer known as "the granu- lar layer of Tomes." Studied with the high power, the appearance will be seen to be caused by irregular spaces in the dentine matrix communicating with the dentinal tubules and filled in life with protoplasm of the fibrils. Compare the dentine in the root with that in the crown (page 171). After studying all the cementum in the three sections, select three fields, one from the gingival, one from the middle, and one from the apical portion of the root, and draw the tissues from the surface of the root to the pulp chamber. Show all the details of structure that can be made out with both low and high powers (Fig. 342). With the high power search the cementum for the record of absorptions which have been refilled by cementum. PERIOD VII Drawings of Typical Cavity Walls. — From the molar or bicuspid section select a field in the region of a groove or pit. Imagine a cavity to be prepared in this position. To help in this, an ink line may be made on the cover-glass by using a fine pen and Indian ink, or ordinary ink to which a little sugar has been added. Xow, using both the high and the low power, study the direction of the enamel rods as they appear in the line of the cavity wall, and make a drawing showing the structural requirements for a good wall in this position. From any one of the three sections select a field in the gingival third of the labial or buccal surface and indicate the line of a cavity wall in the same way. Study with the low and the high powers the direction of the enamel rods as they appear in the line of the walls of the cavity, and make a drawing showing the structural requirements for good walls in these positions (page 80). 440 DIRECTIONS FOR LABORATORY WORK PERIOD vm Outline Drawings from Transverse Sections of the Root. — The ground sections of the root have been prepared and should be brought to the laboratory in solution, ready for mounting. The three sections should be mounted together under one cover-glass, using balsam about the consistence of molasses. The sections may be studied at once, but after the day's work upon them they should have a spring clip adjusted to the cover-glass and be put away until the balsam is thor- oughly hard, otherwise they may work out to the edge of the cover-glass. With the millimeter gauge measure the length and breadth of each section, multiply the measure- ments by twenty, and lay off a rectangle as in making the longitudinal drawings. Draw the outline of the section and the pulp chamber as accurately as possible before studying the section with the inicroscope. With the low power follow the dentocemental junction around each section and draw it into the outline. Fill in half of each section, showing the direction of the dentinal tubules, the position and char- acter of the granular layer of Tomes, the number and posi- tions of the lacunae, and the other structural characteristics of the cementum. In this study the record of the reduction of size of the pulp chamber which may be noted by changes in the direction and the character of the dentinal tubules (page 185). Label the section with the name of the root from which it was ground, your name, and the date. PERIOD IX Study of Secondary Dentine and Cementum. — With the low power find a field where there is a distinct demarcation between dentine of earlier and later formation, and draw it accurately with the high power. Compare the size of the tubules, their number, their direction, and their diameter in the earlier with the later formed dentine; is there any con- GROUND SECTIONS OF BONE 441 nection between the tubules of the two portions? Find a similar field from a longitudinal section and study in the same way, making an accurate drawing. Search all of the ground sections with the low power until a field is found where the dentinal tubules are cut trans- versely. Adjust the high power objective and study the field. Notice that by focussing up and down with the fine adjustment the tubules seem to move in a circle, showing the spiral course through the matrix. Using a red blood corpuscle as a standard, note the size of the tubules, their distribution in the matrix, and the amount of matrix sepa- rating them. Look for the appearance of Newman's sheath, which is that portion of the matrix forming the immediate wall of the tubule. Draw accurately one field as seen with the high powder. Study the cementum from all the ground sections for an area showing absorption and rebuilding, and if found, draw one field with the high power. Draw five or six lacunae with their canahculi as seen with the high power, selecting as great a variety of forms as possible. PERIOD X Ground Sections of Bone.— From a shaft of a femur or humerus saw a disk about one-quarter of an inch thick. In doing this notice the appearance of the marrow cavity especially as you look into it toward the articular ends. Saw the disk into sectors with an arc of about a quarter of an inch on the outer surface. From this piece saw two thin sHces — one at right angles to the axis of the bone, the other parallel with it. These should be ground as directed in the introduction for the grinding of transverse sections of the root, and be brought to the laboratory ready to mount. They should be mounted in hard balsam as described in the mounting of longitudinal sections of the teeth. Label the slide with the name of the bone from which the section is taken and the direction in which it is cut. Study the transverse section with the low power, working out the 442 DIRECTIONS FOR LABORATORY WORK arrangement of the lamellae and the distribution of the subperiosteal and Haversian system bone (p. 252). Draw the tissue from the surface of the bone to the marrow cavity. This drawing should be not more than an inch wide and the full length of the page. With the high power objective and low power eyepiece, draw one or two Haversian systems. Study the arrangement of the Haversian canals as seen in the longitudinal sections. With the high power draw at least three lacunae, showing one cut lengthwise, one trans- versely, and one as seen from above. PERIOD XI Decalcified Bone. — One of the bones from a small animal has been decalcified, embedded, sectioned, and stained with hematoxylin and eosin. Receive from the desk two sections, one of which is cut longitudinally, the other transversely. Mount in balsam in the usual way. Label the slide with the name of the animal, the bone from which it is cut, and the direction of the section. Study the transverse section with the low power, noting the bone corpuscles in the lacunae, the tissue in the Haversian canals, and the marrow. With the high power draw one field showing two or three Haversian systems, one of which has been partially destroyed in the building of another. Draw with the high power one field from the marrow cavity. From the longitudinal section draw, with the high power, one field showing osteoblasts in a medullary space. PERIOD XII Comparative Study of Subperiosteal Bone and Cementum. — For this day's work the previously mounted sections must be used, the longitudinal sections of the teeth, the transverse sections of the root, the ground and decalcified sections of bone. Study the cementum and the subperiosteal bone as shown in these sections and make one drawing of cementum DENTAL PULP FROM UNERUPTED TOOTH OF SHEEP 443 and one drawing of subperiosteal bone to show the com- parison in structure. Compare the regularity in form and arrangement of the lacunae in the bone with the irregularity in form and position of the lacunae in cementum. Note that in the bone the lacunae lie between the layers ; in the cementum they may be between the layers or entirely within a single layer. Compare the regularity in the arrangement and thick- ness of layers with the corresponding irregularity in cementum. Note the size, number, and arrangement of the canaliculi radiating from the lacunae in bone, and compare them with the canaliculi of the cementum. PERIOD xm Dental Pulp from the Unerupted Tooth of a Sheep. — An unerupted molar or premolar of a yearling lamb was removed from the lower jaw by splitting the bone. The pulp was pulled out of the partially formed dentine embedded in paraffin, sectioned, stained with hematoxylin and eosin. Bring to the desk a clean slide with a drop of balsam upon the centre of it and receive a section. Label the slide: " Pulp from unerupted tooth of sheep, stained with hematoxylin and eosin." Study first with the low power. Upon the circumference of the section the layer of odontoblasts may or may not be shown, depending upon whether in the removal of the pulp the fibrils have pulled away from the dentine, or the odontoblasts have been pulled oft' from the surface of the pulp. They are usually present, at least in spots. Note the number and arrangement of the blood- vessels and the distribution of the connective-tissue cells. With the low power draw a portion from the surface to the centre, showing the layer of odontoblasts if present. With the high power draw one field showing a bloodvessel and the connective-tissue cells, taking particular pains to repre- sent their forms correctly. If there are any odontoblasts present draw one field showing them and the layer of Weil (see page 209). 444 DIRECTIONS FOR LABORATORY WORK PERIOD XIV Dental Pulp, Normal Human. — A number of human teeth were cracked immediately after extraction and the pulps removed from the pulp chambers. They were embedded in one block of paraffin, sectioned, stained with hematoxylin and eosin, and are ready to be given out. Bring to the desk a clean slide with a drop of balsam on the centre and receive a section. Label the slide: "Transverse section of pulp from human teeth." There will be several sections in this specimen, each from a separate pulp. With the low power follow the circumference of each section, looking for places where odontoblasts are present. Find the best field in the speci- men and draw the layer of odontoblasts as seen with the high power. Notice the fibrils which have been pulled out of the dentinal tubules projecting from the ends of the odontoblasts. If the section is parallel with the long axis of the cells, they will appear as tall columnar cells with a nucleus in the deeper end. If it is oblique to their axis the layer may appear as two or three layers of oval cells. Just beyond the odontoblasts the layer of Weil Avill be seen, usually appearing as a clearer layer containing few cells and about half as wide as the odontoblasts. Beyond this the connective-tissue cells are thickly placed for a short distance, and still deeper they are more widely scattered and about evenly distributed in the rest of the pulp. With the high power draw one field to show the form of the connective-tissue cells of the pulp. With the low power study the distribution of the bloodvessels in all of the sec- tions. Select the best section and draw the entire section, to show the size, number, and arrangement of the large bloodvessels. With the high power draw a single field, to show accurately the structure of a bloodvessel wall. ENDOCHONDRIAL BONE FORMATION 445 PERIOD XV Dental Pulp, Pathologic Human. — By the cooperation of the man in charge of the extracting room, or an extracting speciahst, teeth with hving but inflamed or h\7)eremic pulps were dropped as soon as extracted into a fixing fluid. The teeth were afterward cracked and the pulps removed, embedded, and sectioned as before. Bring to the desk a clean slide with a drop of balsam on its centre and receive a section. Label the sHde: "Pathologic pulp from human tooth stained T^'ith hematoxylin and eosin." Follow the same routine in studying these specimens as in the case of the normal pulp. It is impossible to tell just what condi- tions will be present. Compare the size and number of the bloodvessels with those in the normal tissue, and the char- acter and distribution of the cellular elements. Look for nodules of calcoglobuU, especially in the inflammatory speci- mens, and make a diagnosis of the condition, as shown in the specimen. See the chapter on the Structural Changes in the Pulp and Pathological Conditions for fuither assistance on the work in this material. PERIOD XVI Endochondrial Bone Formation. — A forming bone from a human fetus has been embedded, sectioned, and stained with hematoxylin and eosin. Receive a section from the desk and mount as usual. Study the specimen -^dth the low power, identifying first the general arrangement of the tissues, following from the unchanged cartilage to the development of bone. Notice the subperiosteal layers on the surface. Make a sketch of a sufficient part of the section to show the changes from the typical hyaline cartilage to the young bone. With the high power draw one field from a primary marrow cavity, showdng osteoblasts laying down lamellse on one of the spicules, and one field showing osteoclasts. 446 DIRECTIONS FOR LABORATORY WORK PERIOD xvn Bone Growth. — A piece of a long bone from a very young animal has been embedded and sectioned transversely to the shaft. Sections have been stained in hematoxylin and eosin, to be mounted as usual. Label the slide: "Growing bone cut transversely, stained with hematoxylin and eosin." Study first with the low power. On the surface of the section will be seen the periosteum, in which the fibrous and osteo- genetic layers can be easily recognized. Bone formation is actively going on, laying down lamella^ under the periosteum which are being transformed into Haversian system bone. With the low power draw a portion of the section from the periosteum to the centre of the bone. With the high power draw a field showing the osteoblasts of the peri- osteum, a field showing the absorption of subperiosteal bone to form a medullary space, and a field showing osteoblasts in a medullary space. PERIOD XVIII Periosteum from Attached Portion. — From a young kitten a portion of a bone in a region to which muscles are attached to the periosteum was carefully dissected out, removing the attached muscle, and the tissue embedded in celloidin, the sections cut parallel to the axis of the bone and perpendicular to its surface. They have been stained in hematoxylin and eosin, and are ready to be given out. Receive a section and mount as usual. Label : " Periosteum from attached portion, stained in hematoxylin and eosin." Study the specimen first with the low power. The outer fibrous layer of the peri- osteum will be seen with the muscle fibers attached to it and the osteogenetic layer with the greater number of cells taking the stain more deeply. Draw with the low power, showing the tissues from the surface of the periosteum well into the substance of the bone. With the high power study the attachment of the muscle fibers to the outer layer of the periosteum, the character and arrangement of the fibers of GINGIVUS AND GUM TISSUE 447 the outer layer, the interlacing of the fibers of the outer and inner layer, the cells, and especially the osteoblasts of the inner layer and the penetrating fibers that are built into the bone. Draw the thickness of the periosteum as seen with the high power, showing the details of structure as accurately as possible. PERIOD XJX Gingivus and Gum Tissue. — The gingivus and gum tissue covering the alveolar process down to the point of reflection on to the cheek was dissected away from the teeth and jaw of a sheep. The tissue was embedded in paraffin and sec- tioned parallel \^'ith the long axis of the tooth. The sections have been stained with hematoxylin and von Gieson, and are ready to mount. Bring to the desk a clean slide ^^'ith a drop of balsam on the centre and receive a specimen. Label the section: "Gingivus from a sheep, stained with hematoxylin and von Gieson." By this staining the cellular elements will have a brownish color, the nuclei dark, the protoplasm lighter, the white fibers should be bright red, and the elastic fibers yellowish. It is a specially good stain for connective tissue. Study with the low power. The epithelial will be stained a brownish yellow or purple. It is a stratified squamous epithelium made up of many layers of cells and with a distinct horny or corneous layer on the surface from the crest of the gingivus to the point where the mucous mem- brane is reflected on to the cheek, or where it ceases to be attached to the gum. This layer is yello\\Tsh in color, and is made up of closely packed scales having no nuclei. They are the remains of epithehal cells from which the protoplasm is gone, leaving only the horny material which it had produced. The portion of the epithelial lining the gingival space has no corneous layer, nuclei being seen in the cells at the surface. The cells are larger and more loosely placed. The connective- tissue papillffi and the projections of epithelium which are between them are extremely long. In the epithelium covering the alveolar process the connective-tissue papilla? 448 DIRECTIONS FOR LABORATORY WORK are broader and not so deep, and the cells are much more compactly arranged. At the point of reflection on the cheek the epithelium changes its character abruptly, the corneous layer disappears, the surface cells showing nuclei, the epi- thelial layer is thicker and made up of larger and more loosely placed cells. This change in the structure explains why the epithelium is easily broken where a movable portion of the membrane passes over the edge of an artificial denture. When an infection reaches the connective tissue a sore is produced that requires some time to heal. Study the connective tissue, which is made up of coarse, wavy bundles of white fibers taking the red stain. In the gum tissue, that is, the portion of the section covering the alveolar process, the bundles are very large and form a very coarse network. Beyond the point of reflection the bundles are finer and more delicate in their arrangement. Elastic fibers take the yellowish stain. Notice the bloodvessels in the connective tissue and the capillaries in the papillae. With the low power draw the entire section so as to show the character of the epithelium and the fibrous tissue in the three parts. With the high power draw the thickness of the epithelium lining the gingival space and at the point where the mem- brane is reflected to the cheek. PERIOD XX Peridental Membrane, Transverse Gingival. — The lower jaw of a young sheep was sawed through between the teeth, cutting the jaw into blocks each containing two teeth. The crowns were broken off or opened so as to admit the fluids to the pulp tissue. The tissues were decalcified, embedded, and sectioned at right angles to the axis of the tooth. They are cut from the gingival portion, and have been stained with hematoxylin and eosin. Receive a section and mount as usual. Label the slide : " Peridental membrane, transverse gingival, stained with hematoxylin and eosin.'' A similar block of tissue preserved in alcohol will be found at the desk. PERIDENTAL MEMBRANE 449 This should be observed so as to study out the relation of the section to the gross appearance of the tissue. Holding the section to the light, observe the distribution of the tissue. Two roots '^'ill be seen cut across. Observe the epithelium on the labial and the lingual, and possibly also that lining the gingival space lying next to the root of one of the teeth. By the aid of the low power sketch the outline of the entire section to show the distribution of the tissues. Note the demarcation where the finer fibers of the peridental membrane unite with the coarser mat of gum tissue. Begin- ning at the centre of the labial surface, follow the fibers springing from the cementum to where they are lost in the gum tissue or attached to the approximating tooth. Draw the portion of the membrane between the two roots, accu- rately representing the arrangement of the fibers. The epi- thelial structures wall be seen lying between the fibers close to the cementum, and should be showm in the drawing (p. 308). With the high power study the cementoblasts and the epithelial structures. Make a dra-^ing of one field, showing all the details of structure as accurately as possible. With the high power draw one field showing the fibrous tissue between the roots and the relation of the fibroblasts to them. This field should include a bloodvessel. PERIOD XXI Peridental Membrane, Alveolar Portion, Transverse. — The sections for this work have been cut from the same block as the preceding, but are in the occlusal third of the alveolar portion and as close to the border of the alveolar process as possible. Receive a section. Mount as usual and label the shde : " Peridental membrane, alveolar portion, transverse, stained with hematoxylin and eosin." Study the general arrangement of the tissues and make a sketch as in the case of the previous specimen. Note the muscle fibers from the muscles of the lip attached to the periosteum on the labial surface of the process, the bone of the labial plate, the septum separating the alveoli, the peri- 20 450 DIRECTIONS FOR LABORATORY WORK dental membrane filling the space between the bone and the surface of the root, the layers of the cementum, the dentine and the pulp. After studying the specimen with the low power as care- fully as possible, draw the peridental membrane surrounding one root, including the thickness of the labial plate of bone with its periosteum and a part of the lingual plate. In this drawing represent accurately the fibers of the peridental membrane, their arrangement in the bundles, and the relation of the bundles to each other and the bloodvessels. To do this the fine adjustment must be used to obtain ideas of the third dimension of space. With the high power draw one field from the wall of the alveolus, showing the attachment of the fibers to the bone, the osteoblasts on the surface of the bone, and the other cellular elements. This field should include a bloodvessel. With the high power draw the thick- ness of the cementum at some point where a specially strong bundle of fibers is attached. This should show the fibers embedded in the cementum, cementoblasts on the surface, and the branching and interlacing of the bundles. PERIOD xxn Longitudinal Section of the Peridental Membrane. — The lower incisor of a young sheep was removed from the jaw by sawing through between the teeth, leaving two teeth in each block. The crowns of the teeth were broken off near the level of the gum so as to admit the reagents to the pulp chamber. The tissues decalcified, embedded in celloidin, and sectioned. They were cut through from labial to lingual, and only the ones from the central portion used. They have been stained in hematoxylin and eosin and are ready to mount. Mount the section as usual and label the slide : " Longitudinal section through the peridental membrane of a sheep, labiolingual, stained in hematoxylin and eosin." First hold the section up to the light and note the relation of the tooth to the bone and the soft tissues. Study the section with the low power and make a sketch showing the TOOTH GERM 451 general distribution of the tissues. Show the pulp chamber, dentine and cementum, bone, periosteum, gum tissue, and epithelium. Do not attempt to fill in the drawing more than diagrammatically, for it would require too much time. The object of the drawing is to get the general relation of the tissue before studying parts of it in detail. Compare the form of the labial and the lingual gingivus and make a draw- ing of the lingual, showing the details of structure as far as the border of the process and as accuratel}^ as possible. With the high power draw the thickness of the epithelium lining the gingival space. Study the fibers in the occlusal third of the alveolar process and make a drawing to represent them accurately, showing the cementum at one side and the bone at the other. The entire length of the root can seldom be got in one section on account of the curve of the tooth, so that the fibers can probably be studied to advantage in the occlusal third of the alveolar process only. Draw one field with the high power showing the bloodvessels. PERIOD xxm Tooth Germ. — The head of an embryo pig was embedded in paraffin and sectioned at right angles to the snout. The sections begin in the region of the incisors and far enough back to cut through the nose cavity. They have been stained in hematoxylin and eosin. Bring to the desk a clean sHde and receive a section. Label the shde: "Tooth germ, stained with hematoxylin and eosin." The general form of the section will depend on the position of the section through the head. At the desk is the head of a similar embrj'o preserved in alcohol. This should be observed so as to determine from the section its relation to the head. By holding the section to the light and the use of the low power, make a sketch of the entire section. Note the epiblast covering the outer surface and lining the nose and mouth cavity. The mass which is to form the tongue lying between the roof of the mouth and the mandibular arch. If the section is in front of the angle of the mouth 452 DIRECTIONS FOR LABORATORY WORK there will be no connection between the upper and lower parts of the section. Notice the separation of the nose cavity into right and left by a septum containing cartilage, and the projections of cartilage from the side walls which will form the turbinate bones. On either side of the septum where it joins the palate will be seen little structures known as Jacobson's organ, which later disappear. Notice Meckel's cartilage in the mesodermic mass of the mandible. In the epiderm of the outer surface the beginning of the formation of hairs are to be seen. With the low power follow the epiderm lining the mouth cavity and look for the tooth germ. In each section there are four chances for tooth germs, one on either side in the upper and lower arches. Select the best one and draw it as seen with the low power. The appearance will depend entirely upon the stage of development. With the high power draw enough of the enamel organ to show the arrangement of the cells in the outer and inner tunics and the stellate reticulum. PERIOD XXIV Tooth Germ. — Sections have been prepared in the same way as in the preceding, but from the head of an older embryo, in which the tooth germs are completely formed and calci- fication is ready to begin. Receive a section, mount, and label as before, and draw the outline of the entire section. Note the changes in form and in the tissue elements from the previous section. Bone formation has begun both in the mandible and the maxilla. The amount and distribution of this should be carefully studied. With the low power draw the entire tooth germ, selecting the most typical one in the section. With the high power draw one field showing ameloblasts, odontoblasts, and a portion of the papillae. Find a field in which bone formation is going on and draw it accurately with the high power. APPENDIX CHAPTEPv I THE GRINDING OF MICROSCOPIC SPECIMENS, USING THE GRINDING MACHINE By G. V. BLACK, M.D., D.D.S., Sc.D., LL.D. The Machine. — The basis of this machine is the larger watchmaker's lathe known as No. 2. It must swing 4 inches, the length of the bed must be 12 inches, and be good and solid. A test should be made of the alignment of the lathe head to see that this is exact. If there is any inac- curacy, another lathe should be selected. The power should consist of one of the largest and strongest electric lathes, or motors, made for the use of dentists. This power should be transmitted to the lathe through an overhead shaft of a length that will give good room to operate the lathe with- out the motor being in the way. A pulley may be placed on the left end of the shaft of the motor on one of the brass carriers for grinding wheels. This pulley should carry a good quarter-inch round leather belt. Its diameter should be 2| inches. The pulley on the right hand end of the shaft above should be 5 inches. This will reduce the speed one- half and double the power. On the left end of the shaft should be placed a copy — reversed — of the pulley on the lathe-head, which has 4 grooves. This gives good varieties of speed with each speed of the motor. Another small pulley will be placed near the centre of the length of the overhead shaft, the purpose of which will be explained later (Figs. 343 and 344). The grinding apparatus is built upon a base fitted to the lathe bed in the same wav as the lathe head, or tailpiece. 454 APPENDIX CHAPTER I Fig. 343 Figs. 343 and 344. — A general view of the grinding machine, showing particularly the arrangement for transmitting the power from the electric motor to the machine that does the work. All of this may be made out by reference to the picture while following the text. The bed of the little lathe on the left hand is \2\ inches long, which gives a good idea of the general dimensions. The water is delivered to the grinding stone from a rubber bag or bucket hung on the frame above through a rubber tube to the metal tube on a movable stand, which may be so placed as to bring the brush at its end against the stone. This stand and brush are better seen in Fig. 344. THE GRINDING OF MICROSCOPIC SPECIMENS 455 Fig 344 456 APPENDIX CHAPTER I It has one main shaft parallel with the lathe bed, in good and sufficient bearings to maintain accuracy of alignment and perfect steadiness for long continued usage (see Figs. 343 and 344). This shaft moves freely lengthwise, or back and forward, while turning slowly in its bearings. On the end of this shaft next to the lathe head — the forward end — there is a larger portion,, or ring, and this end terminates in a threaded nipple, upon which the removable grinding disks are screwed firmly against the face of this larger ring, to secure accuracy of adjustment. The use of these disks will be more fully explained later. On the rear end of this shaft, just back of its rear bearing and abutting against it, a large movable nut is placed. This is provided with a thumb screw by which it is made fast at any point desired. Turning this forward pulls the shaft back from the grinding stone. Turning it backward allows the shaft to move forward against the stone. It has also a finger reaching back over a graduated disk just to its rear. This disk is made fast on the shaft, and the two together constitute the micrometer, by which the thickness to which specimens are ground is measured. The movable nut has 40 threads to the inch. The graduation of the disk is on the same principle as that on the screw calipers used by machinists for fine measurements — one-thousandth of an inch — but as this disk is li inches in diameter, the gradu- ations of thousandths are so wide that one-quarter of one- thousandth may readily be used. It differs in plan, in that both the graduation and the parallel lines are placed upon this disk. On the machinist's micrometer the lines are placed on the shaft and the graduations on the nut. The graduation is read from the side of the finger on the movable nut, and the lines are read from its end. It is a very perfect micrometer (Figs. 345 and 346). The forw^ard movement of the shaft when grinding, and also the pressure exerted upon the stone, are furnished by a tailpiece placed behind it and attached to the lathe bed. This has a plunger actuated by a spiral spring, which pushes the shaft forward against the stone. The amount of pressure THE GRINDIXG OF MICROSCOPIC SPECIMENS 457 exerted in the grinding is controlled by the amount of com- pression of this spring in fixing the piece to the lathe bed. It may be much or little, as desired. Usually very little pressure is used. When the movable nut has come against the frame in which this shaft turns, the machine may con- tinue to run, but the forward movement of the shaft stops and the grinding ceases in consequence. Therefore there is no danger of grinding a specimen thinner than the measure- ment fixed upon. The further arrangement for finding this measurement will be described later. On the rear portion of the graduated disk, or wheel, a portion or space is toothed, and connected with a worm pinion or threaded shaft by which the main shaft is turned in its bearings. A belt is attached over a wheel on the end of this worm shaft, and extends to the third wheel, previously mentioned, on the overhead shaft. When this belt is ad- justed and the motor started, it causes the main shaft in the grinding machine proper to turn slowly on its axis, while being pressed against the stone by the tailpiece. By this arrangement every part of the specimen fixed on the grind- ing disk is brought successively against every part of the rapidly revolving stone, and is cut perfectly level in all of its parts. The Grinding Disks. — The grinding disks are of brass, accurately turned | inch thick, and If inches in diameter. They have a threaded hole i inch deep in the back to fix them to the nipple on the forward end of the shaft of the grinding machine. A machine should have a half-dozen or more of these, lettered or numbered on the edge, so that records of each may be made when measuring preparatory to mounting specimens for grinding. As the mounting of specimens on others of these may proceed while the grinding on one is going on (for the machine, being automatic, needs little attention), this number at the least is necessar}^ for rapid work. The machine may be stopped and the disk removed from the shaft by a few backward turns, the progress of the grinding examined, the disk returned for further grinding, 458 APPENDIX CHAPTER I Fig. 345 Figs. 345 and 346. — The lathe with the grinding machine mounted upon it in position for work. On the left next to the lathe head is the grinding stone surrounded by the spatter guard, which gathers all of the water from the wheel and dehvers it through its hollow post into a rubber tube below the lathe bed, which conveys it to a conveniently placed receptacle. The water comes from a rubber bag or bucket hung on the overhead frame (see Fig. 343) through a rubber tube to the metal tube mounted on a movable stand so that the brush through which it passes may be placed against the stone. The grinding machine proper is secured to the lathe bed by the larger thumbscrew seen below. The point-finder is seen at the foot of the spatter guard, and is secured by the middle thumbscrew seen below the lathe bed. The shaft of the grinding machine (6 inches long) runs through its whole length, but is completely covered in by its housings to protect its bearings from grit, except at its forward end (next to the grinding stone). This part is protected by a swaddle held by a ring, which keeps the working bearing clean. On this end the grinding disk is seen almost touching the stone. The micrometer is on the other end of the shaft next back of the frame of the grinding machine. Next back of this is a toothed wheel made fast to the shaft. This is actuated by the middle one of the belts descending from overhead (Fig. 343, the left hand belt in Fig. 344). This belt passes over a wheel hidden from view and through a small worm shaft turns the main shaft. Pressure for the grinding is supplied by a plunger actuated by a spiral spring seen at the extreme right hand end. THE GRINDING OF MICROSCOPIC SPECIMENS 459 460 APPENDIX CHAPTER I etc., at any time during the progress of the work. The face of the disk, which should be perfectly flat and parallel with the face of the stone, should always be perfectly bright, so as to reflect light through the specimen when it becomes thin. This enables one to judge very closely of the thickness by the eye (after sufficient practice), that sometimes proves a valuable check on the setting of the measurement in the beginning. The Point Finder. — This is a piece of steel one-eighth of an inch thick, fitted to the lathe bed and set against the face of the lathe head, and made fast by a thumb screw passing through the lathe bed from below. It has a strong arm which passes around other fixtures between the lathe head and the forward end of the base of the grinding machine. It is pro- vided with a set screw, by which a range of variation can be made in the distance of the forward end of the frame of the grinding machine from the lathe head. When this is in place and the measurement of a disk has been made and recorded for the grinding of a specimen to a specified thick- ness, the machine may be taken to pieces and set up again and the grinding proceed without fear of disturbing the measurement, so long as the set screw in the point finder is not moved. It is often necessary during grinding to loosen the grinding machine from the lathe bed, slide it back to adjust something, to remove disks for examination of the progress of the work, etc. This point finder, by preserving the distance between the lathe head and the grinding machine, enables one to do this at will, and again find his exact point of measurement simply by sliding the frame of the grinding machine forward against the set screw of the point finder. This little device seems absolutely necessary to the highest usefulness of the machine. Lap Wheels and Grinding Stones. — I began my work of grinding specimens by the use of lap wheels, but soon dis- carded them because they are dirty. They cut much quicker than stones, however, and may be used for the bulk of the work when much grinding of very hard material is to be done. They are not necessary in grinding teeth, bone, etc.. THE GRINDING OF MICROSCOPIC SPECIMENS 461 but in grinding the harder fossils, especially those impreg- nated with the silica, and in some geological work they become necessary. The best lap wheel I have used is an aluminum wheel. Brass or iron will do the work, but aluminum holds the grit better, cuts with lighter pressure, and does the work quicker. In using these I have fed them continuoush' by hand with carborundum powder in soapy water, using a brush. The Stones. — Anyone who is doing much grinding should have a good supply of stones. I have a pair of carborundum wheels, a pair of emery wheels, a pair of India oil stones, and a pair of Arkansas stones. In each of these pairs one is fine and the other coarser grit. Every stone is dressed to a perfect face on the lathe head where it is to do its work, with a black diamond held in the slide rest. These stones, when put in good shape, seem capable of doing an unlimited amount of work. The conditions of the grinding prevents them from getting out of true. All that seems necessary is to roughen them a bit with a picking wheel when they become too smooth to cut well. For this purpose a much smaller picking tool than the smallest sold for the general mechanical uses seems desirable. This picking wheel has sharp teeth of the hardest steel possible on its periphery. It is held in a handle in such form that the wheel is free to turn. In use it is held against the rapidly rotating stone and slowly passed over its entire surface. It may be held in the hand aided by a tool rest, or may be arranged for use in the slide rest, which is the better form for this work. Watering the Stones. — In grinding, the stones are kept wet in running ice water. A balsam that is too soft to hold a specimen for grinding in water at room temperature will hold it perfectly in ice water, because it is much harder when cold. For this purpose, a receptacle for ice is hung on the frame that holds the overhead shaft, and filled with bits of ice and then filled with water. Both the ice and the water must be clean, for the opening in the tube where it passes the valve which regulates the flow is very small, and a 462 APPENDIX CHAPTER I little bit of dirt or trash might stop the flow. In this case the specimen being gronnd would be burned instantly. A bucket, or a large rubber bag, will answer for this purpose. Then an ordinary rubber tube answers to conduct the water. It is best to have this rubber tube to connect with a metal tube mounted on a stand that may be placed in any position wanted to deliver the water to the stone. This metallic tube is provided with a valve for the regulation of the flow. In its final end it should be provided with a brush of rather long bristles, into which the w^ater is delivered and spread upon the stone. This brush is made upon a short tube fitted into the end of the metal tube. To make this brush, first cover the plain part of the small brass tube with thick shellac dissolved in absolute alcohol. Place a layer of the bristles around it and wrap them tightly wath a fine, strong thread. Then place more shellac over this and another layer of bristles. Continue this until the brush is large enough. Then wrap thoroughly with a cord in shellac, let it dry, and then trim it up. Two of these have served for four years of fairly hard usage. Waste Water. — A spatter guard is made by bending a |-inch round brass tube into a circle, the inner diameter of which is the size of the stones used, and brazing the ends solidly together. Then this is fixed in the lathe and one- fourth of its inner circular diameter is turned away. The grinding stones will then go inside this. Then this piece is provided with a foot and hollow post and fitted to the lathe bed with a washer and nut, the same as other pieces are attached. This catches all w^aste water and through a rubber tube attached to the end of its hollow post under the lathe bed delivers it into a receptacle so placed by the table as to receive it. This prevents all of the spattering of water which would be thrown from a rapidly revolving wheel without it. If it should be inclined to run over when a very full stream is wanted, a piece of rubber dam may be stretched over the foot and pulled to its upper end. This may be caught under the guard in fastening it to the lathe bed, and will deliver any overflow into a receptacle placed THE GRIXDIXG OF MICROSCOPIC SPECIMENS 463 to receive it. In this way nothing is wet or spattered with water. Preparation of Material. — In the preparation of material, such as teeth, bone, etc., in histological work of ordinary delicacy, the specimen is first ground fiat on one side by hand on a rough stone 4 inches in diameter, on the motor, and finished perfectly flat on one of the finer stones on the lathe head. The piece is then washed clean and placed in absolute alcohol for a sufficient time to remove all traces of water, or, when cracking or injury from shrinkage is not feared, it may be dried in the warming box. Then when dried and warmed to about 120° F., it is ready to mount with balsam on the grinding disk for grinding. Management of Balsam. — I suppose the management of balsam will always be a difficult problem with many per- sons. ^Nlany, hovrever, learn it quickly. One may take the dry balsam and dissolve it in xylol, and filter it at a high temperature, say 110° or 120° F. Or one may use the prepared balsam for microscopic mountings. In either case it must be evaporated until stiff enough so that it will move rather sluggishlv at 110° F., but will be fluid at 120° or 130° F. Spiders and Dogs. — For using this another bit of apparatus is necessary. A circular piece of steel made flat on the upper surface is mounted on three legs 1| to 2 inches high. The steel disk should have two rows of holes around its periphery, the one row f inch inside the other. A hard rolled tool steel wire, or rod -fo inch in diameter, should exactly fit these holes. These rods should now be bent at right angles with a short nib on the end, bent again at right angles, so that it wdll point downward when the free end of the rod is set into one of the holes. The length between these two angles should vary from f to IJ inches in three dozen or more pieces which should be prepared. The end which goes in the holes should be cut so that it will not quite reach the surface of the table when dropped into the holes with the end of the nib on the surface of the circular plate. These rods are called "dogs" (Fig. 347.) 464 APPENDIX CHAPTER I With this arrangement a warming box arranged with a thermostat to maintain an even temperature, sufficiently Fig. 347 The spider with a grinding disk upon it and a specimen laid on and secured by bent rods called dogs. When these dogs are placed and pressed down through the holes in the disk of the spider, they hold fast. With a little pressure of the finger outward on the end of the rod below the disk of the spider, the dog slips up and is loose. The disk of the spider is three inches in diameter. high to soften the stiff balsam, is used. The specimen, the balsam, the grinding disk, and the "spider" are placed inside, and allowed to rest until they have reached the temperature THE GRINDING OF MICROSCOPIC SPECIMENS 4G5 desired. Then working quickly, a sufficient amount of balsam is placed on the grinding disk, and the specimen laid on it. This should be pressed down until it is seen that all space under it is JSlled with balsam, but no con- siderable excess should be used. It is well if this rest so in the warming box for ten or fifteen minutes for the balsam to soak well into the specimen. Then the grinding disk, with the specimens, should be laid on the spider and one of the dogs dropped into one of the holes in the steel plate, that will bring its nib on to a part of the specimen chosen. Then another, and still another, should be placed, each with its nib on a different part of the specimen, so that every part of it may be pressed flat on the disk. More dogs should be added if necessary. Now each in turn is pressed down a little, one after another, until all are exerting about all the force the spring of the rods will exert without permanently bending them. In this condition the whole thing is again enclosed in the warming box. At this time any number of specimens of teeth or bits of teeth, bone, etc., that the face of the disk will hold may be placed on the disk, and all may be ground together. Four to six lengthwise sections of incisor or cuspid teeth may be placed at once, or eight to twelve cross-sections. It seems to be best practice, however, not to load the disk too heavily. Four lengthwise sections will grind better than six, as a rule. Now, after the loaded disk had remained in the warming box until all balsam that will come has been squeezed out from under the specimens, all excess of balsam should be very carefully removed, or wiped away, close up against the specimens. Nothing clogs a stone and stops its cutting more effectually than balsam smeared over it, and every excess that may come against the stone should be got out of the way. When this is done the whole thing should be returned to the warming box for from, one to four hours, so that it may dry some about the margins at least. Then it may be removed from the warming box and allowed to cool, and 30 4G6 APPENDIX CHAPTER I await convenience in grinding. It should, however, remain secured on the si)i(ler by the dogs if it is to wait more than a few hours, for the (Usi)()yition of dentine to warp in drying may pull some i)art of the specimen from the disk. Under these conditions, two or three days, or a week, will do no harm. When the grinding is completed, the disk is removed from the machine and the specimens flushed with clean water, and dried by the pressure of a soft napkin folded to several thicknesses, or clean pieces of waste cotton fabric may be used. Then the disk with its specimens should be laid in a dish and sufficient xylol added to cover it, and allowed to rest until the balsam has been dissolved and the specimens released. This will usually require from twenty to thirty minutes, or sometimes as much as an hour. When the specimens are very thin they loosen much quicker than when thick. Any material not penetrated by xylol, as silicified petrifactions and stones, require much more time. When the specimens have loosened, they are ready for permanent mounting for microscopic study. Rapidity of Grinding. — In order to make rapid progress in grinding specimens, one should have six to ten grinding disks, nearly as many spiders, and a large supply of dogs. The machine is so nearly automatic in its action that it needs but little w-atching, so that the preparation may be going on while the grinding is in progress. One of the principal points that needs attention is the flow of water. But if the w^ater and ice placed in the receptacle are clean and free from dirt or trash that may stop the flow^ of w^ater, the only care is that the quantity of water is kept up. The vessel should be large enough to hold a supply for several hours. If the stone should run dry, the specimen would be destroyed in a few seconds. Setting the Measurement of Grinding Disks. — When begin- ning any considerable series of grindings, the first thing of importance is to try out and obtain a record of the measure- ments of each grinding disk for the particular stone that may be selected for finishing. I find that most persons, THE GRIXDTXG OF MICROSCOPIC SPECIMENS 467 after some practice, prefer to use a fine stone for the entire grind. In grinding teeth, after roughing down the surface that is to form the specimen, the back is also ground away to a flat surface that will better accommodate the placing of dogs in mounting on the grinding disks. These may be made quite thin and reduce the grinding with the fine stone. Then the stone selected is placed in the lathe head, seeing to.it carefully that the face of the stone is clean. Then the grinding machine is brought up in contact with the set screw of the point finder. The tailpiece is placed in position and pushed up so as to make some pressure on the shaft. Then, with the large nut the shaft is so adjusted that the grinding disk being tried comes close to the stone but does not touch it. Now start the machine and note the running carefully, and while doing so catch the adjusting nut of the micrometer and move it one-thousandth at a time, and listen for the first touch of the disk to the stone. The moment this is heard, quickly reverse the movement of the adjusting nut, and separate the disk from the stone. Try this again and again, until you feel very certain of having detected the first touch of the stone on the disk by moving the adjusting nut half or a quarter of toV"o inch. At last, while it is touch- ing, stop the machine in a position to see the finger on the adjusting nut, and read the measurement and enter it on your record for that disk. In setting for a grind with this disk, turn the adjusting nut so as to draw the grinding disk back from the stone 1-0^7 o" inch. When the specimens to be ground are mounted on this disk, place it back on the machine, start it, seeing that the iced water is running first, and let it run until it ceases to cut, which it will do when the forward movement of the shaft is stopped by the con- tact of the adjusting nut of the micrometer with the rear bearing of the shaft. Then remove the disk and examine the specimens care- fully. If the placement has been accurate, the specimens will be too thick. Replace the disk carefully and turn the nut forward so as to grind one-thousandth of an inch thinner, or one mav do onlv a half of one-thousandth at a time. 468 APPENDIX CHAPTER I Repeat this until the section seems to be thin enough. Then remove and mount the sections and judge them with the microscope. By this time one will have arrived at an accurate measurement of this disk, and the record will be trustworthy for other grinds, and will not have to be repeated until the wearing of the stone begins to leave the specimens a bit thick. Then a half-thousandth of an inch will bring it right. And so on, and on. Each disk will be treated in the same way for each stone used, and if one is doing much grinding all will be running on their records, and all go smoothly. Recently a man who was grinding sections of teeth for me made all of the preparations, preparatory grindings, and disk mounts, ground and removed from the disks ready for mounting forty full-length sections of central incisors in six hours, and had his lunch during the time. Every section was complete, was even in thickness in every part, and all practically the same thickness — a thickness chosen for the special studies in hand. Grinding Frail Material. — While the machine facilitates the production of the more ordinary sections to such a degree as to be indispensable to one having many grindings to do, it is in the production of sections of very frail material that the grinding machine stands out as vastly superior to other methods of grinding. In the study of caries of enamel in which disintegration has rendered the remaining tissue very frail and likely to fall to pieces before it is suffi- ciently thin, we may obtain the required thinness and yet retain all of the tissue. I have also produced exceedingly fine sections of salivary calculus, and equally good sections from small crumbs of serumal calculus. The production of these is slow, but fairly certain of good results. Also in grinding sections of fossil teeth, fossil woods, and the like, in which very fine sections are too brittle to be handled in any way except as stuck to glass, the machine gives excellent results. In geological work it practically removes the difficulties. Good sections of the very brittle stones can be made with fair safety by grinding on the cover- glass. THE GRINDING OF MICROSCOPIC SPECIMENS 469 Plans for Grinding Frail Material. — Much very desirable material for microscopic investigation will be found that is so frail, or at least so brittle, when reduced to sections thin euough for microscopic investigation, that it will crumble to pieces, either in the grinding or in the mounting, by the ordinary processes. For grinding and mounting such material the following processes have been slowly evolved. These may be divided into the balsam process and the shellac process. Such material that, when made fast to a cover-glass and ground in hard balsam, is not liable to go to pieces when this hard balsam is softened by sticking the specimen and glass cover to a glass slide may be ground in hard balsam. If, howveer, the different parts are liable to separate and change position when the balsam softens, shellac should be used for the grinding. I have had some very sorrowful failures in grinding rare specimens of enamel that had no cementing substance between the enamel rods in hardened balsam. For when the softer balsam was added to mount the specimen on the glass slide, the hard balsam was softened and the enamel rods floated out of position. All such material as will not hold together strongly enough to prevent this should be ground in shellac. To grind in hard balsam, the one side of the specimen may be ground flat on the rough stone and then dried out in absolute alcohol. Then the ground side should be saturated to sufficient depth with soft balsam, and laid aside until the balsam has become hard enough to grind smoothl}^ Then the grinding and polishing of this first side should be completed by grinding away all balsam from the immediate surface, and sufficiently into the substance of the specimen to produce a clean, smooth surface of the material. When this has been done, and the surface dried, it should be mounted on an ordinary cover-glass, the thickness of which should have been measured and recorded. In this mounting the cover-glass should be laid on a spider and weight enough placed upon it to insure a perfect fit of the surface of the glass. This should be subjected to about 120° F. heat for from one to five or six hours, for the purpose of expressing 470 APPENDIX CHAPTER I the last bit of balsam possible from between the specimen and the cover-glass. Then it may rest, awaiting the con- venience of the operator, for several days, but the balsam must not be allowed to become "brittle hard," because in that case it loses toughness. All excess of balsam about the margins of the specimen should be carefully removed to facilitate the hardening of that which remains, and espe- cially so that it may not come in contact with the grinding stone, stick to its surface, and interfere with the cutting. Good judgment must be acquired by practice as to the hardening of balsam and shellac in these grinding processes. The best idea of it that can be given in words is this. The balsam or the shellac imist have become firm enough so that it will not drag or allow the specimen to move while grinding in iced water. Neither must it become hard enough to becoine brittle, for then it becomes liable to break. When ready, the specimen is mounted on the grinding disk. This is done by first cleansing the disk, finishing with x;y'lol, and then sealing the cover-glass to this with soft balsam. This should be placed on the spider and well weighted down with dogs. All excess of balsam should be carefully wiped away from the margins of the cover-glass. This may be quickly dried at 120° F., or more slowly at room temperature. It should, however, be warmed for a half hour or more, for the purpose of expressing as much balsam as possible. This cover-glass will be well held for grinding in iced water with only a little drying about the margins, if all excess of balsam is cleaned away closely. The balsam should not become very hard. If the specimen is of considerable bulk and of a quality of material that can be cut with a steel saw, the disk may be caught in a vice "with leather-cushioned jaws to avoid bruising," and the bulk of the material removed with a jeweller's saw, leaving only a moderately thin section for grinding. Or if the material is very hard, as stones, silicified fossils, etc., the disks may be mounted upon the slide rest and cut with the slicing disks, to be described later. The specimen is now ready for the final grinding. The THE GRIXDIXG OF MICROSCOPIC SPECIMEXS 471 record for measurement with the particular stone to be used in finishing has been made, tried out on unimportant material, and the cover-glass has been measured and its record made. With this data, the disk is screwed to its place, the micrometer turned to the proper measurement for the finish, the iced water arranged, the machine set in motion, and it will do the rest. When coarser stones are used for cutting away considerable material, I find those with just a little experience prefer to gauge the amount of the cutting by the eye for the coarse stone. Removal of the Cover-glass from the Disk. — I remove the cover-glass with the specimen from the grinding disk in two different ways, as seems at the time best. First, the grinding disk is placed on a heated piece of metal that will warm the grinding disk quickly. Have a stick of rather soft wood ready, the end of which is cut to a rather sharp angle and thinned down almost in the form of a blade. When the grinding disk begins to warm, catch the margin of the cover-glass with the end of the stick and begin to make steady pressure. As the disk warms, so as to soften the balsam, the cover-glass will begin to move under the steady pressure, slowly at first, but more rapidly later, and will slide oft' the grinding disk before the specimen is loosened. For this plan the cover-glass should be pretty strong, one and one-half to two thousandths of an inch thick. Otherwise there will be great danger of breaking it. It is well in some cases to run just a little xylol around the margins of the cover-glass and partially dissolve the balsam that has become driest before the heating. Great care must be taken not to allow the xylol to spread on to the specimen, for it would loosen it very quickly. The specimen is then turned downward and placed on a tiny drop of balsam on a glass slide, and quickly pressed down close and level. As the new balsam will soften the old, it should not be moved further than quickly to apply a light spring clip to hold it steady. The parts of the speci- men are less likely to move if this is laid on ice for an hour or more. 472 APPENDIX CHAPTER I The Use of Shellac. — In the second plan shellac is used instead of balsam for hardening the specimen and holding its parts together in the first grinding. This part of the work is other\vise done in the same way. The drying of the shellac requires more time usually than the balsam. The attachment of the cover-glass to the grinding disk is done in the same way as when balsam is used to hold the specimen on the cover-glass — that is, with balsam. The grinding proceeds similarly in every respect. In the removal of the cover-glass from the grinding disk, and mounting the specimen, comes the important differences in the two processes. Xylol dissolves balsam very quickly. But xylol does not dissolve shellac at all. Therefore, in- stead of pushing the cover-glass of the grinding disk, the disk is laid in xylol and the balsam dissolved out. In this there is no danger of detaching or moving the specimen if the handling is careful. When cleaned, it is inverted upon a glass slide on a drop of balsam without fear of movement of parts of the specimen, no matter how frail. The Preparation of Shellac. — To keep shellac in condition for this work has some difficulties. The dry scales should be dissolved in absolute alcohol so as to make a moderately thick varnish. It should then be filtered at a temperature of 110° to 120° F., or be made thinner and filtered at room temperature. Great care should be exercised to keep the filtrate from exposure to a damp atmosphere, for it absorbs water readily and then will throw down fine crys- tals, which destroy its value for microscopic purposes. After being filtered it should be evaporated in a close warming box at about 110° to 120° F., to the consistence of syrup. In doing this it is well to divide the supply into two or three grades — a thinner, medium, and a thicker solution. The thinner solution will be used for saturating frail speci- mens before any cutting is done. The thicker solutions for attaching specimens to the cover-glass for grinding. The medium solution for either purpose, as the material may seem to require. THE GRINDING OF MICROSCOPIC SPECIMENS 473 The Grinding from Crumbled Material. — There is often important material for investigation that can be had only in very small crumbs, or broken pieces, such as serumal cal- culus, sands, crumbled bits of strange stones, or mixtures of such material as is found in some of the coarser sands. These, on microscopic investigation, ma^' tell important stories as to their origin and throw important light upon geological questions. In addition to the ordinary microscopic observa- tion, the polariscope may be turned on these, and reveal important facts as to their origin and structure. Also many things will be found in botanical work, such as obtaining sections of small seeds, and the like, which will give important information. Having done a few of these grindings, especially of the very frail dental material, such as serumal calculus, extremely frail fossil teeth, etc., plans of work more or less well adapted have been developed. For instance, I have obtained excellent sections of serumal calculus, which can be had only in small crumbs or flakes, in this wise. A small collection of these bits are first im- mersed for a time in absolute alcohol, or until all air has been removed if they are dry, or if the}' are freshly gathered, until all water has been removed. Then a cover-glass is prepared by covering its central part with the thicker solution of shellac, and these crumbs are placed in this, in what seems to be the best position for obtaining sections. These are allowed to soak full of the shellac, under a close cover, and then uncovered to dry up. Then, if some of the pieces seem to need it, more shellac is added from time to time, until the embedding seems sufficient. This may be dried at room temperature, or in the warming oven at 110° to 120° F. Shellac should not be subjected to much higher temperatures for a considerable time, because continued high temperature for many days together seems to injure the strength. When this is sufficiently hard for smooth grinding, and before it has become too brittle (determining this point re- quires some experience), the preparation is cemented to the 474 ArrENDiX C If AFTER I grinding disk with halsam and ground to such a point as seems most favorable for obtaining sections. This point is to be determined by frequent removal of the disk from the machine and examination of the exposed surfaces of the several pieces. When this })art is done, the cover-glass is dissolved off of the grinding disk by xylol. Then another cover-glass is attached to the surface tvith the least possible amount of shellac. This in turn is dried to the right consistence. Then the last cover-glass placed — that is, the one on the side that has been ground — is secured to the grinding disk with balsam. When this has set it is placed on the machine and the first cover-glass is ground away and the section ground to the required thinness. They are again dissolved off of the grind- ing disk, and may be at once mounted in balsam on the microscopic slide. Difficulties in Grinding. — In the grinding of material enveloped in shellac, or in balsam, either of these materials are apt to gum up the stone and stop the cutting, or render the grinding very slow. When this is from balsam, it may be quickly removed after drying the stone by washing w4th xylol on a brush, or a bit of cloth, while the stone is slowly revolved. When clogged with shellac, the washing is done with absolute alcohol. This requires much more time, and some advantage may be obtained b}^ using pumice stone wuth the cloth or with cork. After rubbing with pumice stone, a very thorough washing with alcohol should be made to remove the last particles of pumice, before re-beginning the grinding. Even with this, the ground surface is apt to be rough or scratched for a time by particles of the pumice lodged on the stone. These will soon disappear, how'ever. Yet the pumice should not be used in the last portion of the grinding. With much grinding of hard substances, the surfaces of the stones become worn so smooth that the}' do not cut well. Then the picking tool should be run over the surface until it is perceptibly roughened. This will cause the stone THE GRIXDIXG OF MICROSCOPIC SPECIMENS 475 to cut briskly for a considerable time, and at first — following such sharpening — the ground surface of the specimen is likely to be full of scratches. In that case a smooth stone should be used for the finishing. Much care should be taken in keeping the stones in good condition. Except in the ways mentioned, no dirt or grit should be allowed to come in contact with their surfaces. A single particle of grit lodged in the surface of the stone will fill the whole surface of the ground section with scratches. Although I shut up my stones in a close fitting drawer, I find it necessary to cover each with a close fitting cloth that is so closely woven as to exclude all dust. In taking care of the machine itself, one cannot be too careful. All of the bearings of the lathe head and of the grinding machine should be swaddled with candle wick saturated with oil to prevent the ingress of gritty particles. This is especially needful when using the aluminum saws and feeding them with carborundum powder. Then every bearing about the whole machine should be especially pro- tected to prevent the possibility of getting grit in the bear- ings. Carelessness in such a matter will quickly ruin a fine bit of mechanism. But with this care, such a machine should continue to do its work well for a lifetime (Figs. 348 and 349). The Slicing Mechanism. — This is an arrangement for slic- ing very hard substances which cannot be cut with the ordinary steel saw — such as the enamel of teeth, silicified fossils, rocks, etc. This consists of an aluminum disk fitted to the lathe head, and surrounded by a special form of spatter guard that admits of the use of the periphery for cutting, and an object holder fixed upon the slide rest of the lathe. The object holder consists of a clamp that grasps a brass tube slotted at the free end in which teeth, or other objects may be made fast with plaster of Paris or sealing wax for slicing. Or in place of this a brass man- dril, upon the end of which there is a threaded nipple by whidi any of the grinding disks may be attached. These are fixed in the position of the ordinary tool post, 476 APPENDIX CHAPTER I and may be swung horizontally to any possible position in relation to the aluminum disk. An object can there- fore be so placed on the disk as to be cut in any direction desired. Usually these are fixed upon the disk with sealing wax. In using the aluminum disk it is fed with carborundum powder suspended in soapy water to give it some stickiness. Fia. 348 Figs. 348 and 3 19. — Arrangement for slicing very hard material. Fig. 348 is the more ordinary view of the machine with the slide rest and object holder in position. In Fig. 349 the lathe is turned about to give a better view of the slide rest, object holder, spatter guard, and aluminum disk. In these illustrations the slotted tube is used (see text) to hold the object being cut. Notice that the disk used for cutting is sur- rounded by a spatter guard which is open for a space at one side so that the periphery of the disk may be used in cutting. This guard gathers all water and grit used in cutting, and delivers it into the pan below through its hollow post. When doing this kind of work all of the bearings of the machine should be carefuUy wrapped (swaddled) to keep them safe from intrusion of grit. This is applied with a brush by hand, and is kept going so constantly as to prevent the disk from running dry. The ordinary aluminum plate, of twenty-four to thirty gauge, may be used for making these. They are first cut in circles THE GRINDING OF MICROSCOPIC SPECIMENS 477 by hand, as large as the lathe will swing (4 inches), and then are cut down to 3 J inches with a tool in the slide rest. These are quickly made when wanted. They wear out rapidly, and yet one of them will do much cutting of very hard substances, and do it accurately and delicately. Rings may readilv be cut from the ordinary test-tubes without special Fig. 349 i I ■eg|L\ \f)h^ ^an -^\-.^^ <^^^^S^ \^Hr LJ* ■^>'> ml- ^_aB»i ■^^j0^^ T in. 000^'n^2 1 ' ^ danger of breaking. The crown of a molar tooth may be cut into many slices; fossil teeth, silicified fossil woods, stones, etc., may readily be sliced as thin as they can be handled in the after-work of preparation. APPENDIX CHAPTER II THE THEORY OF HISTOLOGICAL TECHNIQUE The first requirement of histological technique is to obtain a general view of the theory of procedure. Many beginners make the mistake of supposing that directions for histological technique can be followed like the receipts of a cook book, or the directions for an experiment in chem- istry. This is very seldom the case, and while it is always necessary to follow directions accurately, it is still more necessary to follow them intelligently. All histological methods require judgment. For instance the length of time required for xylol to replace absolute alcohol in a block of tissue which is to be embedded depends upon the size of the piece, the character of the tissue, the temperature, and possibly some other factors. It is therefore impossible to say exactly what time would be required, and the experi- menter must use the judgment which has been acquired as the result of experiment. In the same way no experi- menter can make up a stain and be sure that it will work exactly like the last lot made by the same formula until he has tried it. Even with the same stain the length of time required for staining a section depends upon the thick- ness of the section, the character of the tissue, and the pre- liminary technique it has been through. So that all time directions must be considered as approximate, and to be successful the experimenter must study, first, the object to be obtained by the use of each reagent, and the peculiar action of the reagent upon the tissue. For observation with the compound microscope trans- THE THEORY OF HISTOLOGICAL TECHNIQUE 479 mitted light is ordinarily used. The object must therefore be thin and transparent enough to allow the light to pass through it. The higher the magnification the smaller the field, that is, the smaller the portion of the tissue that can be seen at one time, and the less depth of focus, and con- sequently the thinner the sections must be. A section that would be excellent for study with the f objective may be almost valueless under a yV> ^nd sections that are splendid under the yV might be of little value under the f . In other words, the thickness of the section should be related to the magnification with which it is to be studied, and to the size of the structural elements which make up the tissue. For the study of the organs and tissues of multicellular organisms there are three general methods — (1) teasing, (2) macera- ting, and (3) sectioning. Teasing. — In this method a small portion of the living tissue is torn apart with two needles in a drop of normal salt solution or some indifferent medium which will not aifect the tissue. In this way it is spread into a thin film and squeezed a little between a slide and cover-glass so as to separate the structural elements when they may be directly observed. Of course, in stud^'ing such a preparation it must be remembered that the tissue has been forcibly torn apart and effects of violence must be looked for. These often bring out facts of structure which would not otherwise be as easily seen. After teasing the living tissue, staining agents may be used to facilitate the study of structure. The fresh tissues are often so transparent and made up of substances of so near the same refracting index that very little structure can be made out without the use of staining agents. It must be borne in mind that staining agents are of two classes, diffuse and selective. A diffuse stain gives an even color to all of the tissue and facilitates the study chiefly by rendering it less transparent. A selective stain combines more readily with one portion of the tissue than another, rendering it more conspicuous. Selective stains therefore must be thought of as chemical agents which combine with parts of the cell or tissue and demonstrate 480 APPENDIX CHAPTER II chemical differences in the structural elements. For in- stance, basic anilines react with the chromatin of the nucleus, producing a colored compound. The stain may then be washed out of the section, leaving only the nuclei colored. Acid anilines in general are diffusive stains giving a general color to the cytoplasm. In a similar way certain stains will react only or chiefly with intercellular substances, rendering them more conspicuous. For staining freshly teased specimens methyl green, the formula for which will be found under the paragraph on stains, is an excellent agent. Teased specimens are never very permanent, though they may be preserved for a considerable length of time by mounting in glycerin or glycerin jelly and putting a ring of varnish or white lead around the edge of the cover-glass so as to prevent evaporation. Maceration. — When an organ is composed of more than one tissue the structural elements may be separated by selecting an agent which will act upon one and not upon the others; for instance, the muscle fibers of a voluntary muscle may be separated by treating a piece of tissue with dilute alkali, which w^ill soften and dissolve the connective tissue, allowing the muscle fibers to separate. In a similar way dilute alcohol will soften the cementing substance be- tween the epithelial cells. By first treating a piece of tissue with the proper agent and then teasing, the form of the structural elements of the tissue can be made out. By treating a portion of connective tissue containing both white and elastic fibers with dilute hydrochloric or acetic acid, which dissolves the white fibers, elastic fibers which could otherwise not be seen may be made out. Macer- ating and teasing methods are of great assistance to the study of tissues in sections, and it would be often very difficult to obtain true ideas of structure from sections without their assistance. Sectioning. — For the study of the structural elements in their relation to each other in the tissue sectioning is the one method. As they exist in the body, however, some of the tissues are too soft and others too hard to allow the cutting THE THEORY OF HISTOLOGICAL TECH XI QUE 481 of a thin enough shoe without disturbing the relation of the structural elements. They must therefore be put through rather an elaborate process in which the object of every step must be understood. Dissecting. — First of all, the material for histological work must be absolutely fresh, that is, living. It must be remem- bered that living cytoplasm is chemically different from dead cytoplasm, and as soon as death occurs postmortem changes begin which gradually destroy the structure. The period from death to the beginning of histological methods of preparation should be measured in minutes, not in hours. Tissues that have been dead for a few hours will not react with the staining agents so as to produce the brilliant specimens that can be obtained from fresh material, and often a few days will render material entirely useless except for the grosser anatomical relations. The specimens to be studied should be dissected while the cells of the tissue are still alive, and in doing so the greatest care should be used not to disturb the relation of the tissues. Fixing. — Histologically this word means killing. After dissecting out the tissue to be studied, and while the cells are still alive, it must be immersed in some liquid that will kill the cells and fix their structure as w^hen alive. The pieces should be made small enough for the fixing agent to penetrate them rapidly, and the size of the piece that can be used depends upon the density of the tissue, its character, and the nature of the reagent. Some fixing agents are very much more penetrating than others. All fixing agents coagulate or set the cytoplasm and tend to prevent shrink- age. The success of all the following steps and the value of the specimen for the study of detail of structure depend upon the perfection of fixation. The fixing agents most commonly used are bichloride of mercury, potassium chromate or chromic acid, osmic acid, alcohol, and formalin. The formulas for the same will be found on pages 496 and 499. Hardening. — Since all the fixing agents coagulate living cytoplasm, they are also to a greater or less extent hardening 31 482 APPENDIX CHAPTER II agents., and after fixiii^r tissues may be handled with less danger of disturbing the relation of the struetural elements. Some fixing agents, espeeially chromic fluids, may be con- tinued in their action as hardening agents until the tissue has attained the proper consistency for sectioning, but, as a rule, it is necessary to use other agents for this purpose. In all cases the fixhig agent must he thoroughly trashed out of the tissue before the process is continued. Alcohol is the uni- versal hardening agent, and at the same time it removes the water from the tissue. In carrying tissues from water to alcohol several grades must always be used, and the more delicate the tissue the more gradual must be the changes. If a piece of tissue is taken from water and placed in 95 per cent, alcohol, the diffusing currents will be so strong as to disturb structure and at the same time the hardening action is so energetic as to produce shrinkage. From water a tissue should never be placed in alcohol stronger than 70 per cent., where it should be allowed to remain for twenty- four hours. From 70 per cent, it may be taken to 95 per cent, for the same length of time, and from 95 per cent, to absolute, which will entirely remove the water and prepare the tissue for embedding. If the tissue is very delicate, it should be placed in water, then in 50 per cent, alcohol, and carried through in grades of 10 per cent, to 95 per cent. Embedding. — In order to cut thin sections of tissue the piece must be surrounded and infiltrated with some firm substance which will not only support the entire piece, but will soak through the tissue, filling all intercellular spaces and supporting the individual structural elements. At the same time the embedding material is used to fasten the tissue firmly to a block of fiber or wood which can be grasped in the clamp of the sectioning machine. Two kinds of material are used for this purpose. Substances that are fluid when warm, and solid when cold, as paraffin, or substances which may be dissolved in volatile liquid and are solidified by evapo- ration, as celloidin. In both of these methods the substances, as a rule, are either oily or insoluble in water, and therefore the tissue must be thoroughly dehydrated — that is, have all THE THEORY OF HISTOLOGICAL TECHNIQUE 483 the water removed from it before it is placed in the embed- ding material. To accomplish this there should be at least one change of absolute alcohol. From the absolute alcohol the tissue should be placed in a fluid which is a solvent for the embedding material, so that it will penetrate the tissue more perfectly and rapidly. Heat is always injurious to the tissue, and in embedding in paraffin, therefore, the tissue should be kept in the melted paraffin for the shortest possi- ble time and paraffin of as low a melting point as is consist- ent with sufficient hardness for cutting should be used. In embedding by evaporation the evaporation should not be too rapid or the shrinkage will be increased. Tissues may be kept blocked and ready to cut for a long time, but as a general principle the shorter the time the more perfect will be the specimen. Sectioning. — For sectioning some sort of machine is necessary, and many kinds have been designed, the general principles of which are all the same. They consist of a clamp which holds the knife and a clamp which holds the specimen, and can be adjusted in such a way as to bring the specimen in proper relation to the knife. The position of the specimen is advanced by a micrometer screw so that sections of any desired thickness may be sliced. The delicate part of this machine is the micrometer screw. The essential to the success of its working is the sharpness of the razor, and for such specimens as decalcified bone the razor must be heavy and strong, so that the edge will not spring in cutting the hard tissue. Staining. — The detail of staining process will be described in the next chapter, but it must be remembered that stains, as a rule, are water solutions and the sections must be carried through the grades of alcohol to water before they are ready for the stain. After staining they must be carried back through the grades of alcohol, so as to remove the water entirely before they can be mounted in balsam, which is not soluble in water. Mounting. — Except in serial work, but one specimen should be placed on a slide, and this should be in the centre, leaving 484 APPENDIX CHAPTER II room at either end for a label. In serial work the sections may be placed at one end of the slide, preferably the left hand, leaving room at the right for one label. Labelling. — Nothing in histological technique is more impor- tant than labelling, especially in all research work. Through every step of the process the specimen must be kept track of, and a mixing of labels may spoil months of work. A laboratory notebook containing a record of all material and work should always be on the tables. I have found a system of date and number convenient. For instance, on June 4 a number of specimens are dissected out; in the note- book the record of the source of the tissue is made; the first piece is placed in a bottle of fixing fluid and the bottle labelled 6-4-1911, No. 1; the second, 6-4-1911, No. 2, and so on. In the notebook the description of each block and the date and the hour when it was placed in the fluid is recorded. In this way the tissue may be carried clear through recording each step in the process, and when it is sectioned and mounted we can follow its history in the notebook. Every slide should be labelled first with the date and the block number so as to follow its technique; second, the name of the tissue, and third, the kind of stain- ing. This should be placed on the right hand label, leaving the left hand label for index and file number if the section is preserved. Indexing and Filing. — Many beginners make the mistake of not indexing and filing their slides. They think because they have only a few, that they can easily find anything they want, and that they will wait until they have a larger number before they begin a system, but when a large number have piled up they can never find time to go back and arrange them as they should be. And only one who has failed in this way knows the annoyance of looking through hundreds of slides to find one that he knows he has someplace. APPENDIX CHAPTER III GENERAL HISTOLOGICAL METHODS Fixing. — As has been seen from the preceding chapter, fixing is the first and one of the most important steps in all histological methods. No degree of care in the latter steps can make up for any imperfection in it. As a general statement all fixing agents have advantages and disadvan- tages so that in research work several should be tried and their results compared. For class-room work, however, minute details are not so important. Certain general principles may be stated. Bichloride of mercury is espe- cially adapted to the fixing of epithelium of the mucous membrane. It, however, does not penetrate rapidly, and small pieces must be used. Crystals are liable to form in the tissue, and special precautions must be taken for their removal. Flemming's and Zenker's fluids and the fluids containing osmic acid are used chiefly in research. For classwork the author uses Miiller's fluid and Miiller's fluid and formalin almost entirely. Stains are apt to work better after chromic flxing fluids. The formulas for several of the best fixing agents with directions for their use are found in the last chapter. Washing. — Except for special purposes, fixing fluids are washed out of the tissues in running water, and they should be thoroughly removed. For this purpose the author has made a galvanized iron tank in which a gauze tray divided into small gauze compartments is suspended. The water is brought into the tank through a rubber tube with the mouth resting on the bottom, and leaves through a spout at the top to which another tube can be attached. In this APPENDIX ('II AFTER III way a lar^e number of specimens can be washed at once and their identity followed. Preserving Tissues. — After washing, the tissues should be carried through the grades of alcohol, and may be preserved for a considerable time in SO per cent, alcohol, but it should be changed occasionally. Choice of Sectioning Methods. — The choice between paraffin and celloidin for embedding depends upon the character of the section desired and the nature of the tissue. Small objects and those of delicate structure, such as embryos, dental pulps, etc., are best sectioned in paraffin. Large pieces and blocks containing tissues of different densities are more easily cut in celloidin. Paraffin can be cut much thinner than celloidin, and is therefore preferable for the minute study of cell structures with the high power. Cel- loidin sections are more easily stained and are easier han- dled and therefore preferable for the study of the arrange- ment of tissues with low powers. The author prefers celloidin sections for classwork whenever possible. Embedding in Paraffin. — Tissues fixed and washed are taken from 80 per cent, alcohol and placed in 95 per cent, for twenty- four hours, then in absolute alcohol for the same length of time, and the absolute alcohol should be changed once duiing this period, from absolute alcohol to xylol, in which the tissue should remain until it is clear and translucent. The time in xylol should be as short as possible, as it has a harden- ing action. From xylol it is placed in a solution of paraffin in xylol, and from this to soft paraffin in the paraffin oven, at a temperature of not over 52° or 53° C. In this it should remain from one-half to six hours, when it is transferred to hard paraffin in the oven for the same length of time. The time in the oven should always be as short as is consistent with a perfect infiltration. After sufficient time in hard paraffin the tissue is blocked in the following way: A mould is made by placing L-shaped pieces of metal together on a flat slab. These are manufactured for the purpose. Melted paraffin is poured in the mould and the tissue arranged in it, placing it so that the sections will cut in the GENERAL HISTOLOGICAL METHODS 487 direction desired. A film of paraffin will harden at once on the slab and the tissue can be placed very nicely with the needles. As soon as a film has formed over the surface the slab with the mould should be immersed in cold water, so as to harden the paraffin as quickly as possible. When cold, sections may be cut at once or the block may be preserved in a pasteboard carton properly labelled. As a rule, paraffin sections should be cut as soon as possible. Paraffin. — The paraffin for embedding sections must be of the best quality. That prepared for this purpose by Griibler is preferable. It should be of two grades, that melt- ing at 45° C, and that melting at 54° C. The hard paraffin is mixed with the softer, so as to give a melting point at about 52°. In winter softer paraffin should be used than in summer, as the cutting quahty depends upon the adjust- ment of the paraffin to the temperature of the room. If the paraffin is too hard the sections are liable to tear and curl; if it is too soft, the structure of the tissue will be disturbed in cutting. Perfect infiltration is always necessary for good sections. Chloroform or oil of cedar may be substituted for xylol in this process. Xylol is most rapid, but has some disadvantages in its action on the tissues, especially if left too long. Cutting Paraffin Sections. — If the specimen has been placed at one end of the block, the other end of the paraffin may be clamped in the microtome. If the piece is too small, it should be fastened to a block of vulcanized fiber with melted paraffin and the fiber block clamped in the specimen holder. With a sharp scalpel the excess of paraffin around the specimen should be trimmed off, leaving the block in a rectangular form. The microtome knife is placed at right angles to the microtome bed, and the side of the block should be parallel with the blade. The specimen should be brought up just to the edge and the first section cut. The knife should be moved with a quick, sharp motion, as par- affin sections are chopped when the knife is in this position. The knife is pushed back, the block lifted with the micro- meter screw so as to give a section of the proper thickness, 488 APPENDIX CHAPTER III and the second section cut. If the paraffin is of the proper consistency and the block has been properly trimmed, the edge of the second section will stick to the first and the sections stretch out over the knife in a ribbon. The ribbons may be transferred to a piece of clean white paper and com- plete series of sections cut. When series are not required larger specimens are often cut better by placing the blade of the knife obliquely and drawing it with a slow, even motion through the block. If the sections show a tendency to roll up when the corner of the section begins to curl over the edge of the knife, it may be caught with the tip of a camel's-hair brush and so section after section transferred to the paper. Paraffin sections should cut at a thickness of from seven to ten microns, but sections as thin as one micron may be cut from small blocks under ideal conditions. Handling of Paraffin Sections. — For staining, paraffin sec- tions must be fastened to the slide or cover-glass. If a few sections are to be cut the slide is preferable; if man}^ sections, as in the preparation of class work, square cover-glasses should be used. In either case the glass must be absolutely clean. A stock of perfectly clean slides and cover-glasses should always be kept on hand (see p. 496). A thin film of albumin fixative is spread upon the glass; this film must be as thin as possible. The best way to spread it is to put a drop of fixative on a glass slab or an ordinary slide, touch the edge of the drop with the end of the little finger and spread it over the cover-glass, wiping off all that can be removed with the finger. Lay the cover-glasses film side up on a piece of paper until the required number have been pre- pared. As each section is cut it is laid on a cover-glass, straightened, and pressed dow^n with a camel's-hair brush. If the sections curl or wrinkle they should be floated on water warmed just enough to soften the paraffin but not melt it. As each section is cut it should be dropped on the top of the water, where it will straighten out. When a number have been placed on the surface of the water they may be picked up by holding the cover-glass in the point of the pliers and slipping it underneath the section and GENERAL HISTOLOGICAL METHODS 489 lifting it as on a section lifter. The water is drained off and the cover-glass placed in the groove of the tray of a IMoore's staining dish/ shown in Fig. 350. Each tray will hold about thirty cover-glasses. They must now be thoroughly dried by leaving them over night at room temperature or for a shorter time in a warm oven, which should not be hot enough to melt the paraffin. When dry, each cover-glass should be picked up in the pliers and passed quickly through the middle of a Bunsen flame, so as to coagulate the albumin, or they may all be fixed at once in an oven. Heat that will just melt the paraffin will coagulate the albumin and hold the section on Fig Soo the glass. By means of a little wire , — ^ --.^ basket the tray with the thirty cover- (1^^^^ glasses may now be carried from X^^J^F^ one dish to another through the fol- ^^^--^ *- ^ lowing necessary reagents. First, a ^ ^==^^^ minute or two in xylol to remove the Morris staining dish. paraffin; then absolute alcohol, then 70 per cent.; then water; Delafield's hematoxylin for five minutes; distilled water to wash off the stain; acid alcohol (70 per cent, alcohol to which 2 or 3 drops of hydrochloric acid has been added to every 100 c.c. of alcohol); again washed in tap water to remove and neutralize the acid (some prefer alcohol to which a few drops of ammonia have been added); 70 per cent, alcohol; eosin for thirty seconds; 70 per cent, alcohol, then 95 per cent., then absolute, and finally xylol. From the xylol the sections may be mounted or given out to the class. For class work a student brings to the desk a clean slide with a drop of balsam on the centre and receives a section. Summary of Paraffin Method. — Tissues in 80 per cent, alcohol. 95 per cent, alcohol, twenty-four hours. Absolute alcohol (changed once), twenty-four hours. Xylol, one-half to six hours. 1 These are manufactured by Bausch &, Lomb. 490 APPENDIX CHAPTER III X3I0I and paraffin, one-half honr. Soft paraffin, one-half to six hours. Hard paraffin, one to six hours. Block. Section. Fix on glass. Heat. Xylol, one minute. Absolute alcohol, one minute. 95 per cent, alcohol, same. 70 per cent, alcohol, same. Distilled water. Hematoxylin, five to ten minutes. Tap water. Acid alcohol. Tap water or ammonia alcohol. 70 per cent, alcohol. Eosin, thirty seconds. 70 per cent, alcohol. 95 per cent, alcohol. Absolute alcohol. Xylol. Mount in balsam. Label. Celloidin Method. — Tissues fixed and washed are taken from 80 per cent, alcohol and placed in 95 per cent, for twenty-four hours; then in absolute alcohol for the same length of time, changing the alcohol once. Then into a mixture of absolute alcohol and ether for twenty-four hours, from this into a thin solution of celloidin, in which they should remain for from two days to a week. From the thin solution they should be placed in a thick celloidin solution, about the consistency of syrup, for the same length of time. The tissues may be kept in the celloidin solution indefinitely without injury, and if the tissue is difficult to infiltrate it may be of advantage to leave them in these solutions for weeks or months. In this case the bottles must of course be perfectly corked to prevent evaporation. GEXERAL HISTOLOGICAL METHODS 491 Blocking of Celloidin Material. — There are several methods for blocking celloidin materials, of which the author prefers the following: Thick celloidin is poured into a stender dish or a small Petrie dish until there is enough to abundantly cover the specimens, which are arranged on the bottom of the dish. A match or bit of cork is placed under the edge of the cover so as to allow slow evaporation. In a day or two the celloidin will attain the consistence of a thick jelly. A knife is now passed around each tissue and the celloidin containing the specimen lifted out, and the excess of celloidin is trimmed away. A vulcanized fiber block has one surface dipped into the thick celloidin and the specimen arranged upon it. Thick celloidin is now added to surround and cover the tissue with its adherent celloidin. As soon as this is hardened so as to form a film it is dropped into 80 per cent, alcohol to harden the entire mass. In this it must remain at least twenty-four hours before it can be sectioned. Tissues embedded in celloidin may be kept for years in 80 per cent, alcohol blocked and ready to cut without great injury to the tissue. Celloidin solutions for embedding should be kept in two grades and labelled "thick" and "thin" celloidin. The latter should be quite fluid, the former about a syrup consistence. Scherring's celloidin is furnished in two forms, in shreds and granules. The former will dissolve more rapidly. About half an ounce is placed in a large- mouthed bottle, and a mixture of equal parts of absolute alcohol and ether added. It dissolves slowly and should be shaken frequently. When this solution is sufficiently thick, part may be poured into another bottle and diluted with sufficient absolute alcohol and ether for the thin solution, while the thicker portion is poured into a bottle for the thick solution, and absolute alcohol and ether may be added to the stock bottle to dissolve the residue. When blocking tissues as described above the trimmings are dropped back into the stock bottle. Cutting Celloidin Sections. — The fiber block is clamped in the specimen holder and adjusted. The knife is set diago- 492 APPENDIX CHAPTER III nally so as to cut with a drawing motion, and both the knife and the block are kept flooded with 80 per cent, alcohol. The sections may be allowed to pile up on the knife, and after eight or ten are cut they are slid off with a camel's-hair brush on to a section lifter and transferred to 80 per cent, alcohol, in which they may be kept for some time. Staining Celloidin Sections. — For transferring celloidin sec- tions the most convenient thing is a small tea-strainer with a handle. These may be got for a few cents at any hardware store. By means of this the sections are transferred to 70 per cent, alcohol, from this to distilled water, and are stained from five to ten minutes in Delafield's hematoxylin. The stain is then washed off with tap water, destained with acid alcohol, w^ashed in tap water or ammonia alcohol, stained thirty seconds in eosin, washed with 70 per cent, alcohol, from this to 95 per cent., in which they should be given two or three changes. From this they are transferred to beech- wood creosote or some other clearing agent (see p. 503), and in this they may be kept until they are ready to mount or to be given out to the class. For class w^ork the student brings to the desk a clean slide, and a section is placed upon the centre of it. After blotting off the excess of oil he adds a drop of balsam, covers with a cover-glass, and labels the specimen. Summary of Celloidin Method. — Tissues in 80 per cent, alcohol. 95 per cent, alcohol, twenty-four hours. Absolute alcohol, changed twice, twenty-four hours. Absolute alcohol and ether, twenty-four hours. Thin "celloidin, two days to a week. Thick celloidin, the same. Evaporate. Block. 80 per cent, alcohol to harden or store. Sections cut in 80 per cent, alcohol. 70 per cent, alcohol, one minute. Distilled water. Hematoxylin, five to ten minutes. GENERAL HISTOLOGICAL METHODS 493 Tap water. Acid alcohol. Tap water or ammonia alcohol. 70 per cent, alcohol. Eosin one minute. 70 per cent, alcohol to wash. 95 per cent, alcohol, changed twice. Creosote. Mount in balsam. Label. Serial Sections with Celloidin. — It is difficult to cut series of sections with the celloidin method. The simplest process and one used with success is to carry the sections in order from the knife to the slide, arranging three or four at one end of it and leaving room for a label. Strips of porous tissue paper are cut the proper size and one laid over the sections to hold them in place. A thread is then lightly wrapped around the slide and paper, when they may be carried through the necessary agents for staining, in Naples jars. After they are cleared the paper is removed, the excess of the oil blotted off, the balsam put upon the section and covered with a long cover-glass. SPECIAL METHODS Dental Pulp. — The unerupted premolars from a young sheep furnish excellent material for the study of the dental pulp. The jaws of sheep slaughtered for spring lamb can be easily obtained from the stockyards, and while still warm are placed in JNIiiller's fluid and formalin, in which they are taken to the laboratory. The temporary incisors are still in place and may be used for peridental membrane material. With the bone forceps the cortical plate is removed and the unerupted teeth dissected from their crypts. By grasping the base of the dental papillae with the pliers the pulp may be pulled out of the dentin. They should then be replaced in Muller's fluid and formalin for twenty-four hours when 494 APPENDIX CHAPTER III tlu'v may be carried through the usual process, embedded ill ])araffin, and sectioned. Human Pulps. — By the cooperation of the extracting room human })ulps for liistoh)gical work may be obtained. As soon as extracted the tooth should be wrapped in a gauze napkin, placed in the jaws of a heavy vise, which is carefully tightened until the tooth cracks. The same thing may be accomplished by a heavy hammer on an anvil. A few trials of this will enable one to crack the tooth so that the pulps may be easily remo\'ed without injury. The cracked tooth is put in Miiller's fluid and formalin for twenty-four hours, when the pieces of dentine are removed and the pulp care- fully lifted out of the pulp chamber. It is then carried through the regular process, embedded in paraffin, and sectioned. If the teeth are not perfect clinical history should be noted. Periosteum. — Young kittens that have not attained their full growth may be used for this purpose. The bone should be very carefully dissected so as not to injure the periosteum and then sawed in pieces, using a fine metal saw. It is usually best simply to saw it in two at the middle of the shaft and to fix it in Miiller's fluid and formalin. After fixing and washing, it should be cut in small pieces and decalcified in 2 to 5 per cent, nitric acid. A comparatively large volume of acid should be used and a pad of cotton placed in the lower half of the bottle, or the tissue suspended by a thread. It is best to change the acid once a day. Decalcification may require from two days to a week, and should be tested by passing sharp needles through the tissues. As soon as decalcified the tissue should be washed for twenty-four hours in running w^ater, carried through the grades of alcohol, and embedded in celloidin. The sections should be cut at right angles to the shaft. Peridental Membrane. — For class work the peridental mem- branes of sheep are the best for study, as their fibers are large and their direction easily observed. They are much better than those of either cat or dog, in which the fibers are much finer and the bone more dense. The jaws are brought from GENERAL HISTOLOGICAL METHODS 495 the stockyards in ]\Iiiller's fluid and formalin, the crowns broken ofl' at the level of the gum so as to expose the pulp chamber, and the jaws sawed through so as to leave two teeth in each block, after which they are replaced in Miiller's fluid and formalin for two days, decalcified in nitric acid, and thoroughly washed. They may now be cut into small blocks for transverse sections and embedded in celloidin. Embryological Material. — For the study of the tooth germ in class work embryo pigs of all ages are easily obtained. The entire embryo should be at once placed in Miiller's fluid or a saturated solution of picric acid and water. In ^Miiller's fluid they should remain a week; in picric acid, forty-eight hours. After fixing, the heads are cut ofi", thor- oughly washed, carried through the grades of alcohol, and embedded in paraffin. APPENDIX CHAPTER IV FIXING AGENTS AND STAINING SOLUTIONS Cleaning of Slides and Cover-glasses. — Slides or cover- glasses on which paraffin sections are to be mounted must be absolutely clean. They should be dropped in strong sulphuric acid and allowed to remain a few minutes. The acid should then be poured off and thoroughly removed with water, and strong acetic acid poured on. After remaining a few minutes wash the acid off thoroughly and wipe from alcohol. Keep ready for use in a clean box. Meyer's Fixative. — The white of an egg is chopped with a pair of scissors and filtered through muslin, diluted with an equal volume of glycerin, and a little sodium oxalate added to prevent decomposition. FIXING AGENTS Flemming's Solution. — A good solution for fixing nuclear structures is the chromic-acid solution of Flemming: Parts. Osmic acid, 1 per cent, aqueous solution 10 Chromic acid, 1 per cent, aqueous solution 25 Glacial acetic acid, 1 per cent, aqueous solution 10 Distilled water 55 Small pieces are fixed in a small quantity of the fluid for at least twenty-four hours. They are then washed for the same number of hours in running water and passed through 50, 75, and 80 per cent, each twenty-four hours into 90 per cent, alcohol. FIXIXG AGEXTS 497 A stronger solution is made as follows: Part3. Osmic acid, 2 per cent, aqueous solution 4 Chromic acid, 1 per cent, aqueous solution 15 Glacial acetic acid 1 Fol's Solution. — A modification of Flemming's solution: Parts. Osmic acid, 1 per cent aqueous solution 2 Chromic acid, 1 per cent, aqueous solution 25 Glacial acetic acid, 2 per cent, aqueous solutioa 5 Distilled water 68 Corrosive Sublimate. — An excellent fixing fluid is made by saturating distilled water with corrosive sublimate. Small pieces about 0.5 cm. in diameter are immersed in this fluid for from three to twenty-four hours, then washed in running water for twenty-four hours, and then transferred into 70 per cent, alcohol. After twenty-four hours the tissues are placed in SO per cent, for the same length of time and then preserved in 90 per cent. It often occurs that after changes in temperature crystals of sublimate are formed on the surface or in the interior of the object. For their removal a few drops of iodine and potassium iodide are added to the alcohol (P. Mayer). It is a matter of indifference whether the 70 per cent., SO per cent., or 90 per cent, alcohol is thus iodized. In future treatment of the object, as well as in sectioning, any such crystals of sublimate will not be found to be a hindrance. In the case of delicate objects it is better to undertake their removal after sectioning by adding iodine to the absolute alcohol then used. Acetic Sublimate Solution. — An excellent solution specially used for embryonic tissues and for organs containing only a small quantity of connective tissue. To a saturated aqueous solution of sublimate, 5 to 10 per cent, of glacial acetic acid is added. After remaining two to three hours or more in this solution, the objects are transferred to 35 per cent, alcohol and then passed through the higher grades of alcohol. 32 498 APPENDIX CHAPTER IV Picric Acid. — Small and medium-sized objects (up to 1 c.c.) are fixed in twenty-four hours in a saturated aqueous solution of picric acid (about 0.75 per cent.). Objects of considerable size may be left in this solution for weeks with- out detriment. The tissues are then transferred to 70 or 80 per cent, alcohol, in which they remain until the alcohol is not colored by the picric acid. Instead of a pure solution of picric acid, the picrosulphuric acid of Kleinenberg, or the picric acid of P. INIayer may be used. Picrosulphuric acid is made as follows: 1 c.c. of concentrated sulphuric acid is added to 100 c.c. of a saturated aqueous picric acid solution. Allow this to stand for twenty-four hours and dilute with double its volume of distilled w^ater. The picric acid solution is made by adding 2 c.c. of pure nitric acid to 100 c.c. of saturated picric acid solution. Filter after standing for twenty-four hours. Chromic Acid. — Chromic acid is used in a | to 1 per cent, aqueous solution. Small pieces are fixed for twenty-four hours, larger ones for a longer time. The quantity of the fixing fluid should equal at least more than fifty times the volume of the tissues to be fixed. After fixing, objects must be w^ashed for at least tw^enty-four hours in running water,' then through the grades of alcohols, and preserved in 80 per cent. Two to 3 drops of formic acid to every 100 c.c. of chromic acid solution improve their fixing properties. MuUer's Fluid. — Potassium bichromate 2 to 2.5 grams Sodium sulphate 1 gram Water 100 c.c. This solution requires a long time for fixing, at least several weeks, and for large pieces several months. During the first few weeks the solution should be changed every three or four days and later once a week, until it remains clear. Tissues should be thoroughly washed in running water at least tw^enty-four hours. For some special purposes it is better to wash in alcohol. Tissues should be carried through the grades and preserved in 80 per cent, alcohol. FIXING AGENTS 499 Wliile tissues are in ^Nliiller's fluid they should be kept in the dark. Miiller's Fluid and Formalin. — Muller's fluid 100 c.c. Formalin 10 c.c. The addition of formahn to Miiller's fluid greatly hastens fixation. It is an excellent agent of great penetrating power, and tissues stain very well after it. Twenty-four hours will fix tissues of ordinary size, though they may be left longer without damage. Bone fixed too long in formalin is liable to be hard to cut. Zenker's Fluid. — Grams. Potassium bichromate 2.5 Sodium sulphate . 1.0 Corrosive sublimate . 5.0 Glacial acetic acid . . .5.0 Water 100.0 Add the glacial acid in proper proportion to the quantity of the solution to be used, and not to the stock solution. x'Vllow the tissues to remain in this solution for from six to twenty-four hours. Then wash in running water for from twelve to twenty-four hours and transfer to gradually concentrated alcohol. Crystals of sublimate which may be present are removed with iodized alcohol. Zenker's fluid penetrates easily and fixes nuclear and protoplasmic struc- tures equally well without decreasing the staining qualities of the elements. Formalin. — Of recent years formalin, which is a 4 per cent, solution of the gas formaldehyde in water, has been much used as a fixing fluid. ^Nlake a solution by adding 10 parts of formalin to 90 parts of water or normal saline solution. Small pieces of tissue should remain in this for from twelve to twenty-four hours, larger pieces a number of days or weeks, and then transfer to 90 per cent, alcohol. 500 APPENDIX CHAPTER IV STAINING AGENTS Delafield's Hematoxylin. — Hematoxylin crj'stals 4 grams Absolute alcohol 25 c.c. Ammonia alum, aqueous solution 400 c.c. Methyl alcohol 100 c.c. Glycerin 100 c.c. Dissolve hematoxylin crystals in absolute alcohol and add to the alum solution, place in an open vessel for four days, then filter and add the methyl alcohol and glycerin. Hemalum (Mayer, 91). — One gram of hematin is dissolved by heating in 50 c.c. of absolute alcohol. This is poured into a solution of 50 grams of alum in 1 liter of distilled water and the whole well stirred. A thymol crystal is added to prevent the growth of fungus. The advantages of hemalum is as follows: The stain may be used immediately after its preparation, it stains quickly, never overstaining, especially when diluted with water, and penetrates deeply, making it useful for staining in bulk. After staining sections or tissues are washed in distilled water. Safranin. — Safranin 1 gram Absolute alcohol 10 c c. Aniline water 90 c.c. Aniline water is prepared by shaking up 5 c.c. to 8 c.c. of aniline oil in 100 c.c. of distilled water and filtered through a wet filter. Dissolve the safranin in the aniline water and add the alcohol. Filter before using. Stain sections fixed in Flemming's solution for twenty- four hours and decolorize with a weak solution of hydro- chloric acid in absolute alcohol (1 to 1000). After a varying period of time, usually only a few minutes, all the tissue elements will be found to have become bleached, only the chromatin of the nucleus retaining the color. STAINING AGENTS 501 Methyl Green.— Stains very quickly. One gram is dis- solved in 100 c.c. of distilled water to which 25 c.c. of absolute alcohol is added. Rinse the sections in water, then place in 70 per cent, alcohol for a few minutes, transfer to absolute alcohol for a minute, etc. Hematoxylin. — Van Gieson's Acid Fuchsin-Picric Acid Solu- tion. — Stain in any of the hematoxylin solutions, and after rinsing sections in water counterstain in the following: Acid fuchsin, 1 per cent, aqueous solution 5 c.c. Picric acid, saturated aqueous solution 100 c.c. Dilute with an equal quantity of water before using. The hematoxylin stained sections remain in the solution from one to two minutes, are then rinsed in water, dehydrated, and cleared. Hematoxylin-Eosin.— Sections already stained in hema- toxylin are placed for two to five minutes in a 1 to 2 per cent, aqueous solution of eosin or in a 1 per cent, solution of eosin in a 60 per cent, solution of alcohol. They are then washed in water until free from the stain, after which they remain for a short time in absolute alcohol. In place of the eosin solution a 1 per cent, aqueous solution of benzopurpurin may be used for the following solution of erythrosin (Held). Erythrosin 1 gram Distilled water 150 c.c. Glacial acetic acid 3 drops Silver Nitrate Method. — Especially useful for staining inter- cellular substances of epithelium, endothelium, and meso- thelium, and the ground substance of connective tissues. It may be used on either fresh or fixed tissues, fresh tissue, however, being more satisfactory. Spread the tissues to be stained in thin layers; immerse in a 0.5 to 1 per cent, solution of silver nitrate from ten to fifteen minutes; rinse in distilled water and place in fresh distilled water or 70 per cent, alcohol or a 4 per cent, solution of formalin and expose to direct sunlight until they assume a brown color. The sun- light reduces the silver in the form of fine particles which 502 APPENDIX CHAPTER IV appear black on being examined with transmitted light. The preparations thus obtained may be examined in glycerin or dehydrated and mounted in balsam. Glycerin. — To mount in glycerin transfer the sections from water to the slide, cover with a drop of gl^xerin, and apply the coverslip. Sections colored with a stain that would be injured by contact with alcohol and where clearing is not especially necessary are mounted this way. Farrant's Gum Glycerin. — In place of pure glycerin the following mixture may be used: Gljcerin 50 c.c. Water .50 c.c. Gum arabic (powder) . . .50 grams Arsenous acid 1 gram Dissolve the arsenous acid in water. Place the gum- arabic in a glass mortar and mix it with the water, then add the gl^'cerin. Filter through a wet filter paper or through fine muslin. To preserve such preparations for any length of time the cover-glasses must be so fixed as to shut off the glycerin from the air. For this purpose cements or varnishes are used, by painting over the edges of the cover-glass. These masses adhere to the glass, harden, and fasten the cover-glass firmly to the slide, hermetically sealing the object. Kronig's is one of the best formulas for varnish, and is made as follows: Melt 2 parts of wax and stir in 7 to 9 parts of colophonium and filter the mass hot. Before employing an oil immersion lens it is best to paint the edges with an alcoholic solution of shellac. Silver Nitrate. — In thin membranes and sections the vessel walls can be rendered distinct by silver impregnation, which brings out the outlines of their endothelial cells. This may be done either by injecting the vessel with a 1 per cent, solution of silver nitrate, or with a 0.25 per cent, solution of silver nitrate in gelatin. This method is of advantage, since after hardening the capillaries of the injected tissues appear slightly distended. Organs thus treated can be sectioned, but the endothelial mosaic of the vessels does not appear definitely until the sections have been exposed to sunlight. STAINING AGENTS 503 The injections of lymph channels, lymph vessels, and lymph spaces is usually done by puncture. A pointed cannula is thrust into the tissue and the syringe empties by a slight but constant pressure. The injected fluid spreads by means of the channels offering the least resistance. For this pur- pose it is best to use aqueous solution of Berlin blue or silver nitrate, as the thicker gelatin solutions cause tearing of the tissues. ■ Clearing Agents. — Clearing agents are substances of high refracting index, mostly oils, which are used to displace alcohol and prepare tissues for embedding and sections for mounting in balsam. Clearing agents for embedding in paraffin must be miscible with alcohol and solvents for paraffin. They are called clearing agents because the tissues become translucent and clear in them. Xylol is the most rapid and probably most used agent. It has, however, a hardening action on the tissues, especially if they remain too long in it. Pure oil of cedar wood when free from turpentine is an excellent agent. Chloroform has been largely used for the same purpose. Before celloidin sections are mounted in balsam they must be cleared. For this purpose an oil that will mix with 95 per cent, alcohol is desirable, as absolute alcohol softens the celloidin. The oil used must not dissolve the celloidin, and should not dissolve the stain. Beechwood creosote is an excellent agent, and has been largely used. It clears sections rapidly from 95 per cent, alcohol. Oil of bergamot is an excellent agent, also oil of origanum; but in the latter the oleum origani cretici and not the oleum origani gallici must be used. A mixture of equal parts of oil of bergamot and beechwood creosote has been used satisfactorily, and is an excellent agent. A cheaper mixture is made of equal parts of phenol, oil of origanum, and oil of cedarwood. INDEX Absorptiox of roots of temporar}- teeth, 302 Acetic acid and sublimate for fixing, 497 Alveolar process, 379 Analog}', 22 Attachment of teeth, 271 by ankylosis, 274 in fibrous membrane, 272 by hinged joint, 273 by insertion in a socket, 277 B Balsam, 463 management of, for grinding sec- tions, 469 Bichloride of mercury for fixing, 497 Blocking celloidin material, 491 Bone, 247 arrangement of lamellae of, 252 canaliculi of, 249 cancellous, 251, 254 compact, 252 corpuscles of, 248 decalcified, 442 definition of, 247 formation of, 255 endochondrial, 255, 445 endomembranous, 258 growth of, 260, 446 Haversian canals of, 253 system of, 250 influence of mechanical condi- tions on, 380 lacunae of, 249 Bone, matrix of, 247 structural elements of, 247 subperiosteal, 250 varieties of, 250 Branchial arches, 355 Calcoglobulix, 231 Calculus, grinding of sections of, 473 Cell division, 337 indirect, 338 theorv, 336 walls of plants, 238 Celloidin, blocking of, 491 cutting of, 491 method, 490 summary of, 492 sections of, serial, 493 staining of, 492 stock solutions of, 491 Cement corpuscles, 296 Cementoblasts, 295 Cementum, 188 absorption of, 200 canaliculi of, 194 cement corpuscles of, 195 distribution of, 33 embedded fibers of, 196 function of, 29, 189 Haversian canals in, 188 histogenesis of, 189 lacunae of, 194 lamella? of, 190 structural elements of, 190 Chromic acid for fixing, 498 Cleaning slides and cover-glasses, 496 506 INDEX Cleaning agents, 503 Cleft palate, 361 Connective tissues, 240 chemical relations of formed material to cytoplasm, 245 mutations of, 240 relation of, to mechanical conditions, 245 Corrosive subUmate, 497 Creosote, 503 Cutting celloidin sections, 491 paraffin sections, 487 Decalcified bone, 442 Delafield's hematoxylin, 500 Dental foUicles, 364 ligament, 285 papilla, 363 pulp, bloodvessels of, 209 cells of, arrangement of, 209 connective tissue of, 207 definition of, 201 degeneration of, 229 from unerupted tooth of a sheep, 443 function of, 29 sensory, 202 vital, 201 hard formations in, 235 histogenesis of, 203 human, normal, 444 pathological, 445 hyperemia of, 219 infarction of, 224 intercellular substance of, 209 nerves of, 214 nodules in, 229 odontoblasts, 204 pathology of, 219 preparation of, method of, 493 structural elements of, 203 ridge, 362 tissues, 28 distribution of, 30 in adaptation, 35 Dentine, caries of, 157 chemical composition of, 169 dentinal fibrils, 176 distribution of, 32 Dentine, function of, 29, 167 granular layer of, 178 histogenesis of, 167 interglobular spaces in, 179 lines of Schreger in, 184 matrix of, 168 secondary, 184 sheath of Newman, 169 tubules of, 171 diameter of, 171 direction of, in crown, 171 in root, 174 Dermal scales, 22, 271 Development, beginning of calci- fication, 366 chronology of, 372 of dental follicle, 364 papilla, 363 ridge, 362 of enamel organ, 362 of permanent molars, first, 370 second, 371 third, 371 of tooth germ, 362, 364 for permanent teeth, 365 Dissecting, 481 Drawings, 427 of teeth, 425 surfaces, 429 of typical cavity walls, 439 Embedding, 483 in paraffin, 486 Embryology, 335 biological considerations fund^ mental to, 335 branchial arches, 355 chemical ideas related to, 339 early stages of, 340 fertilization, 343 formation of germ layers, 347 frontonasal process, 357 maturation, 340 neural canal, 352 preparation of material, 495 relation of cell theory to, 336 segmentation, holoblastic, 345 mammalian, 348 meroblastic, 348 INDEX 507 Embryology, separation of nose and mouth cavities, 360 spermatogenesis, 340 stomodium, 3.57 , transmission, 338 Enamel, action of acid in caries of, 150 stages in, 153 areas of weakness for cavity margins, incisors, 136 marginal ridges, 127 simple proximal cavi- ties in bicuspids and molars, 137 tips of cusps, 124 atrophy of, structural effects of, 160 bands of Retzius, 60, 115 chemical composition of, 39 cleavage of, 73 cutting of, instruments for, 76 developmental lines in, 122 differences between rods and cementing substance, 46 from other calcified tissues, 38 distribution of, 30 effect of caries beginning in natural defect, 143 on smooth surfaces, 145 intensity and liability, 148 secondary or backward de- cay, 146 on structure of, 143 of structure on cutting of, 56 etching of, 48 function of, 28 gnarled, 54 growth of cap of, 108 lines of Schreger in, 64 mottled, 165 occlusal grooves in, 115 origin of, 38, 362 planing of, 76 refracting index of rods and cementing substance, 51 relation of, to formation tissue, 41 relative solubility of rods and cementing substance, 46 rods of, 43 short, 45 Enamel, straight, 53 stratification of, 60 striation of, 57 structural elements of, 43 form of, 42 walls, structural requirements of, 80 bevel of cavosurf ace angle, 87 classes of cavities, 87 gingival third cavities, 101 incisor pits, 105 in simple occlusal cavities, 90 steps in preparation of, 89 support of marginal rods, 85 of worn surfaces, 86 supported on sound den- tine, 80 white spots in, 162 Endoskeleton, 19 relation of nervous system to, 22 Epiblast, 347 Epithelial structure in peridental membrane, 307 arrangement of cells in, 308 distribution of, 307 Etching and mounting ground sec- tions, 429 Exoskeleton, 19 relation of, to nervous system, 22 Farrant's gum glycerin, 502 ' Fastening teeth to grinding disks, j 465 ; Fertihzation, 343 ' First permanent molars, origin of, 370 Fixative for paraffin sections, 496 Fixing, 481 agents, 496 Flemming's solution, 496, 497 Forces influencing bone growth, 393 I Formalin for fixing, 499 508 INDEX Formalin for teeth, 424 Frontonasal process preserving fluid for Growth of jaws, relation of first molars, 398 357 tissue changes in, 413 of mandible, 375 of membrane bones, 261 Gum glycerin, 502 GiNGivus, gum tissue and, 447 support of, 291 Gland of Serres, 310 Glycerin for mounting, 502 Granular layer of Tomes, 178 Grinding of crumbled material, 473 difficulties in, 474 disks, 457 of frail material, 468 in hard balsam, 469 machine, 453 grinding of sections on, 463 of microscopic sections, 453 description of machine, 453- 457 fastening teeth to grinding disks, 465 frail material, 468 grinding disks, 457 lap wheels and stones, 460 management of balsam, 463 measurement of sections, 467 point finder, 460 process of grinding, 462 rapidity of grinding, 466 removal of cover-glass from disk, 471 spatter guard, 462 spiders and dogs, 463 waste water, 462 watering stones, 461 stones, 460 clogging of, 474 of tooth sections, 425 Ground sections of bone, 441 Growth force, 392 of jaws, 390 eruption of temporary teeth, 394 growth of air space in nose, 412 importance of proximal con- tact, 405 influence of permanent incisors and cuspids, 403 Hair, teeth and, comparison of origin of, 25 of structure of, 24 Hardening, 481 Hemalum, 500 Hematoxylin, Delafield's, 500 eosin and, 501 Van Gieson's, 501 Histological technique, theory of, 478 Holoblastic segmentation, 345 Homology, 22 Hyperemia of dental pulp, 219 acute, 220 chronic, 222 Hypoblast, 347 Indexing and filing, 485 Infarction of dental pulp, 223 Inflammation of dental pulp, 224 Intercellular substances, 236 kinds of, 238 relation of cells to, 237 Isolated enamel rods, 435 Jaws, growth of, 390 Labelling, 485 Laboratory, manner of working in, 427 Lap wheels, 460 INDEX 509 M Maceratiox, 480 JMandible, growth of, 375 Maturation, 3-40 Membrana eboris, 207 Merkel's cartilage, 367 Methyl green, 501 Meyer's fixative, 496 Morris' staining dish, 489 Mounting, 483 Mouth eavitv, 323 epithelium of, 323 mucous membrane of, 323 nerve endings in, 327 submucosa of, 325 taste buds, 331 tongue, 327 muscles of, 328 papillae of, 329 Mucous membrane of mouth, 323 Mailer's fluid, 498 formahn and, 499 Odontoblasts, 204 Oil of bergamot, 503 of origanum, 503 Osteoblasts, 297 Osteoclasts, 299 absorptions by, 300 in burrowing canals, 302 Outline drawings of ground sec- tions, 432 from transverse sections of root, 440 Par AFFIX, cutting of, 487 embedding in, 486 kinds of, 487 method, summary of, 489 sections, staining of, 488 Pathology of dental pulp, 219 Peridental membrane, 279 absorption by, 300 arrangement of fibers of, 283 bloodvessels of, 313 Peridental membrane, cellular ele- ments of, 294 cement corpuscles, 295 cementoblasts in, 295 changes in, with age, 319 definition of, 279 divisions of, 280 epithelial structure in, 306 fibroblasts in, 294 fibrous tissue of, 283 functions of, 281 longitudinal sections of, 450 nerves of, 318 nomenclature of, 280 osteoblasts of, 297 osteoclasts of, 299 practical considerations of, 321 preparation of material, 494 principal fibers of, 283 relation of cementoblasts to cure of pockets, 297 structural elements of, 281 transverse alveolar, 449 gingival, 447 Periosteum, 262 attached, 264, 267, 446 complex, 270 simple, 268 classification of, 262 definition of, 262 functions of, 262 layers of, 265 macroscopic appearances of, 263 preparation of material, 494 relation of attachment of, to burrowing pus, 264 unattached, complex, 265 simple, 265 Picric acid, 498 Placoid scabs, 22, 28, 271 Point finder, 460 Preparation of dental pulp mate- rial, 493 of embryological material, 495 of grinding material, 463 of peridental membrane mate- rial, 494 of periosteum material, 494 of shellac for grinding sections, 472 Preserving tissues, 486 510 INDEX Rapidity of grinding, 466 Reattachment of tissues to surface of root, 297 Relation of nucleus to cytoplasm, 337 of section to crown, 424 of teeth to bone, 374 to development of face, 374 Removal of cover-glass from grind- ing disk, 471 S Safranin, 500 Schreger's lines in dentine, 184 Secondary dentine and cementum, study of, 440 Sectioning, 480, 483 methods, choice of, 486 Segmentation, 345 holoblastic, 345 mammalian, 348 meroblastic, 348 Serial sections with celloidin, 493 Sheaths of Newman, 169 Silver nitrate, 501 injection, 502 Slicing mechanism, 475 Spatter guard, 462 Spermatogenesis, 340 Staining, 483 agents, 500 celloidin sections, 492 of fresh tissues, 479 of paraffin sections, 488 Stomodium, 357 Structure of mandible and maxilla, 377 distribution of bone in alveolar process, 379 in mandible, 384 in maxilla, 390 Structure of mandible and maxilla, influence of mechanical condition in evolution of, 380 Subperiosteal bone, 250 and cementum, comparative study of, 442 Taste buds, 331 Teasing, 479 Teeth, attachment of, 271 chisel, 36 for grinding sections, 423 grinding, 36 relation of, to bone, 27 to exoskeleton, 22 temporary, absorption of roots of, 302 Tissue changes in the physiolog- ical movements of teeth, 413 Tongue, 327 muscles of, 328 papilla of, 329 taste buds of, 331 Tonsils, 331 lingual, 332 palatine, 334 pharyngeal, 334 Tooth germ, 364, 451, 452 for permanent teeth, 365 Transmission, vehicle of, 338 Transverse sections of roots of teeth, 426 W Washing, 485 Watering the stones, 461 Zenker's fluid, 499 ■- -^^ \.itl'x' COLUMBIA UNIVERSITY LIBRARIES i This book is due on the date indicated below, or at the 1 expiration of a definite period after 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