Ultrastructural study of the root dentine surface resuming resorption on human deciduous teeth

Resorption of deciduous teeth is not continuous, but alternates with periods of repair or rest. Dentine surfaces in periods of rest or repair resume resorption by odontoclasts during physiological root resorption of the deciduous teeth. However, no observations of such dentine surfaces have been shown. The characteristic feature of the dentine surfaces resuming resorption remains unknown. Tartrate-resistant acid phosphatase activity (TRAP) was detected on human deciduous teeth. The root resorbing surfaces on these teeth were photographed with a whole-mount light microscope, and the photographed areas were serially sectioned into 0.5 micron semithin sections. Preodontoclasts and odontoclasts were three-dimensionally reconstructed. On root resorbing surfaces, areas with small scattered TRAP-positive cells were observed among areas with many TRAP-positive resorbing odontoclasts and TRAP-negative areas. The sections showed that areas with small scattered TRAP-positive cells have features similar to those of TRAP-negative areas, but there were three kinds of characteristic TRAP-positive cells: preodontoclasts, odontoclasts forming small lacunae, and preodontoclasts, and odontoclasts with cytoplasmic processes extending to the dentine surface, which is covered with cells. These results suggest that the areas with small scattered TRAP-positive cells could be at the stage of resuming resorption, and show that the presence of preodontoclasts and odontoclasts with cytoplasmic processes extending to the covered dentine surface is a characteristic feature of the dentine surface at this stage.     

Development of coronal cementum in hypsodont horse cheek teeth

The horse is a grazing herbivore whose cheek teeth are hypsodon; that is, they possess long crowns that are completely covered by coronal cement at eruption. For elucidation of the sequential events in the formation of this coronal cementum in the mandibular horse cheek teeth, in the present study the lower 3rd permanent premolar teeth (PM₄) from 3.5‐, 4‐, and 5‐year‐old horses were compared by using radiography, microcomputed tomography (Miro‐CT), light microscopy (LM), and scanning electron microscopy (SEM). The present study clearly showed that prior to coronal cementogenesis tartrate‐resistant acid phosphatase (TRAP)‐positive odontoclasts resorbed on the enamel surface of the reserve crown in horse cheek tooth. Enamel resorption areas were relatively narrow, and started from the cuspal tips, and moved in the apical direction during tooth development. A primary cementum was initially deposited on the irregularly pitted enamel–cementum junction (ECJ) of the infolding and peripheral enamel. The infolding cementum filled grooves completely by the time of tooth eruption. On the other hand, in the peripheral cementum, the secondary and tertiary cementum layers were sequentially deposited on the primary cementum. These two cementum layers were sites for the insertion of the periodontal ligaments, and were continually laid down on the primary cementum coronally rather than apically throughout the life. The results of the present study suggest that the coronal cementum of horse cheek teeth is a multistructural and multifunctional tissue, meeting the requirements of its many different functions. Anat Rec, 297:716–730, 2014. © 2014 Wiley Periodicals, Inc.     

Localization of Cathepsin D in Human Odontoclasts. A Light and Electron Microscopical Immunocytochemical Study

Odontoclasts are dentine and cementum resorbing cells whose relationship to bone resorbing osteoclasts is not clear. Like osteoclasts, they possess different cathepsins which are involved in mineralized tissue degradation during the tooth root resorption process in deciduous teeth. Whether cathepsin D, which in osteoclasts probably functions as an activator of other cathepsins, can be found in odontoclasts, has, however, not been investigated before, In order to determine its occurrence and localization, cathepsin D immunocytochemistry was applied to paraffin-embedded sections from 30 human deciduous tooth roots undergoing resorption. Using immunogold postembedding immunocytochemsitry on LR-Gold embedded specimens, the distribution of cathepsin D was investigated at the ultrastructural level. We identified tar-trate-resistent acid phosphatase-positive mono- and multinuclear odontoclasts near and on the periodontal surfaces of tooth roots. Nearly all of these cells showed cytoplasmic granular cathepsin D immunoreactivity. At the electron microscopical level, gold labelling was seen on vacuoles and vesicles of the odontoclasts, which were identified as secondary lysosomes and phagosomes. Extracellularly it was seen along the ruffled border and in neighboured resorption areas of dentine and cementum. These findings indicate that cathepsin D is secreted into the resorbing area of human odontoclasts in order to participate in degradation of mineralized tooth matrix, but may also function as an activator of other proteases in lysosomal organelles.     

Localization of cathepsin D in human odontoclasts. a light and electron microscopical immunocytochemical study

Odontoclasts are dentine and cementum resorbing cells whose relationship to bone resorbing osteoclasts is not clear. Like osteoclasts, they possess different cathepsins which are involved in mineralized tissue degradation during the tooth root resorption process in deciduous teeth. Whether cathepsin D, which in osteoclasts probably functions as an activator of other cathepsins, can be found in odontoclasts, has, however, not been investigated before. In order to determine its occurrence and localization, cathepsin D immunocytochemistry was applied to paraffin-embedded sections from 30 human deciduous tooth roots undergoing resorption. Using immunogold postembedding immunocytochemsitry on LR-Gold embedded specimens, the distribution of cathepsin D was investigated at the ultrastructural level. We identified tartrate-resistent acid phosphatase-positive mono- and multinuclear odontoclasts near and on the periodontal surfaces of tooth roots. Nearly all of these cells showed cytoplasmic granular cathepsin D immunoreactivity. At the electron microscopical level, gold labelling was seen on vacuoles and vesicles of the odontoclasts, which were identified as secondary lysosomes and phagosomes. Extracellularly it was seen along the ruffled border and in neighboured resorption areas of dentine and cementum. These findings indicate that cathepsin D is secreted into the resorbing area of human odontoclasts in order to participate in degradation of mineralized tooth matrix, but may also function as an activator of other proteases in lysosomal organelles.     

Cellular events at the onset of physiological root resorption in rabbit deciduous teeth

For elucidation of how physiological root resorption of deciduous teeth is initiated, the cellular events that occur surrounding the root of rabbit deciduous teeth before and at the onset of physiological root resorption were observed by means of light and electron microscopy. In addition, the cytodifferentiation of odontoclasts during the initial phase of this root resorption was evaluated by histochemical staining of tartrate‐resistant acid phosphatase (TRAP) activity as a marker odontoclasts and their precursors. The present investigation was focused on the physiological root resorption of the deciduous lower second molar of rabbits from Day 0–5 postnatally. At birth, the deciduous molar had not erupted yet, and no TRAP‐positive cell could be found surrounding the tissue adjacent to the root of the deciduous tooth. TRAP‐positive mononuclear cells were initially detected in the coronal portion of the dental follicle of the permanent tooth at Day 1 postnatally. Ultrastructurally, these mononuclear cells had moderate numbers of mitochondria and short‐strand rough endoplasmic reticulum, as well as scattered free ribosomes throughout their cytoplasm. TRAP‐positive mononuclear cells then appeared in the cementoblast layer immediately adjacent to the surface of the deciduous roots. These mononuclear cells projected cytoplasmic extensions between the cementoblasts and made contact with the cementum. At that time, cell–cell contact was frequently observed between these mononuclear cells and cementoblasts. During 3–5 days postnatally, the number of TRAP‐positive multinucleate odontoclasts on the root surface gradually increased. They had well‐developed ruffled borders and made typical resorption lacunae on the root surface of the deciduous tooth. During this early postnatal period, neither inflammatory cells nor necrotic tissue could be observed surrounding the deciduous root. This study demonstrates that the dental follicle of the permanent tooth as well as the connective tissue adjacent to the deciduous root might play important role in site‐ and time‐specific recruitment, development, and activation of odontoclasts before and at the onset of physiological root resorption. Anat Rec 264:387–396, 2001. © 2001 Wiley‐Liss, Inc.     

Ultrastructural features of odontoclasts that resorb enamel in human deciduous teeth prior to shedding

Three dental hard tissues, i.e., cementum, dentin, and enamel, are resorbed by multinucleated cells referred to as “odontoclasts.” These cells have morphological and functional characteristics similar to those of bone-resorbing osteoclasts. However, concerning enamel resorption, which is a process that may occur during tooth eruption, satisfactory ultrastructural data on odontoclastic resorption are still lacking. Ultrastructural and histochemical characteristics of odontoclasts resorbing enamel of human deciduous teeth prior to shedding were examined by means of light microscopy and transmission and scanning electron microscopy. Odontoclasts that that resorbed enamel were tartrate-resistant acid phosphatase (TRAP)-positive multinucleated giant cells that were essentially the same as those that resorbed dentin and cementum. Ultrastructurally, they had numerous mitochondria, lysosomes, and free polysomes in their cytoplasm. In addition, they were characteristically rich in large cytoplasmic vacuoles containing enamel crystals in the cytoplasm opposite the ruffled border. Although they extended a well-developed, ruffled border against enamel surface, a clear zone—an area typically devoid of organelles—was rarely seen in these cells. In many cases, the cells were in very close contact with the enamel surface by the peripheral part of their cytoplasm. The enamel prisms at the resorption surface contained more loosely packed and electron-lucent enamel crystals compared with those of unresorbed, intact enamel. Furthermore, numerous thin needle- or plate-like enamel crystals that were liberated from the enamel matrix were found in the extracellular channels of the ruffled border and in various-sized cytoplasmic vacuoles in their cytoplasm. The superficial layer of the enamel matrix undergoing odontoclastic resorption stained positively with toluidine blue and for TRAP activity. The results of the present study suggest that odontoclasts resorbing enamel secrete acids as well as organic components, including hydrolytic enzymes, into the resorption zone underlying their ruffled border and that they phagocytose crystals that have been liberated from the partially demineralized enamel matrix by acids, subsequently dissolving them intracellularly. Anat. Rec. 252:215–228, 1998. © 1998 Wiley-Liss, Inc.     

Immunolocalization of vacuolar-type H+-ATPase, cathepsin K, matrix metalloproteinase-9, and receptor activator of NFkappaB ligand in odontoclasts during physiological root resorption of human deciduous teeth

To investigate the cellular mechanisms of physiological root resorption in human deciduous teeth, the authors examined the immunocytochemical localization of vacuolar-type H+-ATPase, a lysosomal cysteine proteinase, cathepsin K, matrix metalloproteinase-9 (MMP-9), and receptor activator of NFKB ligand (RANKL) in odontoclasts. H+-ATPase, cathepsin K, and MMP-9 are the most important enzymes for decalcification of apatite crystals and degradation of type-I collagen. In addition, RANKL is one of the key regulatory molecules in osteoclast formation and functions. Odontoclasts developed extensive ruffled borders and clear zones apposed to the resorbing root dentine surfaces. On immunoelectron microscopy, the expression of vacuolar-type H+-ATPase was detected along the limiting membranes of pale vacuoles and the ruffled border membranes of odontoclasts. Cathepsin K in odontoclasts was localized within pale vacuoles, lysosomes, the extracellular canals of ruffled borders, and the underlying resorbing dentine surfaces. MMP-9 localization in odontoclasts was similar to those of cathepsin K. RANKL was detected in both mononuclear stromal cells and odontoclasts located on resorbing dentine surfaces. These results suggest that (1) odontoclasts are directly involved in decalcification of apatite crystals by active extrusion of proton ions mediated by H+-ATPase and (2) extracellular degradation of dentine type-I collagen by both cathepsin K and MMP-9, and (3) odontoclast differentiation and activity are regulated, at least in part, by RANKL, possibly produced by mononuclear stromal cells and odontoclasts themselves in the resorbing tissues. Thus, the cellular mechanisms of physiological root resorption appear to be quite similar to those of osteoclastic bone resorption.     

Cellular events at the onset of physiological root resorption in rabbit deciduous teeth.

For elucidation of how physiological root resorption of deciduous teeth is initiated, the cellular events that occur surrounding the root of rabbit deciduous teeth before and at the onset of physiological root resorption were observed by means of light and electron microscopy. In addition, the cytodifferentiation of odontoclasts during the initial phase of this root resorption was evaluated by histochemical staining of tartrate-resistant acid phosphatase (TRAP) activity as a marker odontoclasts and their precursors. The present investigation was focused on the physiological root resorption of the deciduous lower second molar of rabbits from Day 0-5 postnatally. At birth, the deciduous molar had not erupted yet, and no TRAP-positive cell could be found surrounding the tissue adjacent to the root of the deciduous tooth. TRAP-positive mononuclear cells were initially detected in the coronal portion of the dental follicle of the permanent tooth at Day 1 postnatally. Ultrastructurally, these mononuclear cells had moderate numbers of mitochondria and short-strand rough endoplasmic reticulum, as well as scattered free ribosomes throughout their cytoplasm. TRAP-positive mononuclear cells then appeared in the cementoblast layer immediately adjacent to the surface of the deciduous roots. These mononuclear cells projected cytoplasmic extensions between the cementoblasts and made contact with the cementum. At that time, cell-cell contact was frequently observed between these mononuclear cells and cementoblasts. During 3-5 days postnatally, the number of TRAP-positive multinucleate odontoclasts on the root surface gradually increased. They had well-developed ruffled borders and made typical resorption lacunae on the root surface of the deciduous tooth. During this early postnatal period, neither inflammatory cells nor necrotic tissue could be observed surrounding the deciduous root. This study demonstrates that the dental follicle of the permanent tooth as well as the connective tissue adjacent to the deciduous root might play important role in site- and time-specific recruitment, development, and activation of odontoclasts before and at the onset of physiological root resorption.     

Immunolocalization of vacuolar‐type H+‐ATPase,cathepsin K,matrix metalloproteinase‐9, and receptor activator of NFkB ligand in odontoclasts during physiological root resorption of human deciduous teeth

To investigate the cellular mechanisms of physiological root resorption in human deciduous teeth, the authors examined the immunocytochemical localization of vacuolar‐type H⁺‐ATPase, a lysosomal cysteine proteinase, cathepsin K, matrix metalloproteinase‐9 (MMP‐9), and receptor activator of NFKB ligand (RANKL) in odontoclasts. H⁺‐ATPase, cathepsin K, and MMP‐9 are the most important enzymes for decalcification of apatite crystals and degradation of type‐I collagen. In addition, RANKL is one of the key regulatory molecules in osteoclast formation and functions. Odontoclasts developed extensive ruffled borders and clear zones apposed to the resorbing root dentine surfaces. On immunoelectron microscopy, the expression of vacuolar‐type H⁺‐ATPase was detected along the limiting membranes of pale vacuoles and the ruffled border membranes of odontoclasts. Cathepsin K in odontoclasts was localized within pale vacuoles, lysosomes, the extracellular canals of ruffled borders, and the underlying resorbing dentine surfaces. MMP‐9 localization in odontoclasts was similar to those of cathepsin K. RANKL was detected in both mononuclear stromal cells and odontoclasts located on resorbing dentine surfaces. These results suggest that (1) odontoclasts are directly involved in decalcification of apatite crystals by active extrusion of proton ions mediated by H⁺‐ATPase and (2) extracellular degradation of dentine type‐I collagen by both cathepsin K and MMP‐9, and (3) odontoclast differentiation and activity are regulated, at least in part, by RANKL, possibly produced by mononuclear stromal cells and odontoclasts themselves in the resorbing tissues. Thus, the cellular mechanisms of physiological root resorption appear to be quite similar to those of osteoclastic bone resorption. Anat Rec 264:305–311, 2001. © 2001 Wiley‐Liss, Inc.     

Scanning electron microscopy of the apical structure of human teeth

The objective of this research was to evaluate, by scanning electron microscopy, the apical structure of extracted human permanent teeth with different degrees of pulp and periapical pathology. A total of 25 teeth were extracted: 5 teeth with vital pulp (group I); 10 teeth with pulp necrosis without radiographically visible periapical lesion (group II); 10 teeth with pulp necrosis with radiographically visible periapical lesion (group III). The root apex was sectioned and processed for scanning electron microscopy. In groups I and II, fibers covered the root cementum and there was no cementum resorption or microorganisms. There were areas of cementum resorption in group III with microorganisms on the root apex surface (biofilm) and no fibers. The authors conclude that the presence of chronic periapical lesions causes severe changes in the apical structure with a destruction of fibers and different degrees of cementum resorption forming lacunae in which bacterial biofilm persisted.     

Immunohistochemical Localization of Components of the Insulin-Like Growth Factor-System in Human Deciduous Teeth

To investigate the occurrence of components of the insulin-like growth factor (IGF) system during the resorption process of shedding human deciduous teeth, we investigated sections of 13 decalcified and paraffin-embedded deciduous teeth immunohistochemically with antibodies against IGF-I and -II, six IGF binding proteins (IGFBPs 1-6) and the IGF receptors IGFIR and IGF2R. The teeth were in different stages of resorption and all showed reparative cementum formation. It was found that acellular extrinsic fiber cementum, reversal lines and reparative cellular intrinsic fiber cementum were immunoreactive for both IGFs and various IGFBPs. Therefore, in human deciduous teeth, all subgroups of cementum, but not dentine, may represent sources of components of the IGF system. Odontoclasts did not carry IGFs or the IGF1R, but IGFBPs and the IGF2R. Therefore, these cells, in contrast to osteoclasts, may not respond to IGFs, but may be involved in the release and sequestration of IGFs from cementum during the resorption process. In contrast to odontoclasts, cementoblasts and periodontal ligament (PDL) fibroblasts carried IGF1R. The influence of the IGF system on the function of these cells with respect to periodontal matrix turnover and cementogenesis is discussed. On the behalf of the IGFBP immunoreactivities found, the PDL extracellular matrix can be considered to be a reservoir for IGF system components, where binding proteins may regulate IGF distribution and activity.     

Immunohistochemical localization of components of the insulin-like growth factor-system in human deciduous teeth

To investigate the occurrence of components of the insulin-like growth factor (IGF) system during the resorption process of shedding human deciduous teeth, we investigated sections of 13 decalcified and paraffin-embedded deciduous teeth immunohistochemically with antibodies against IGF-I and -II, six IGF binding proteins (IGFBPs 1-6) and the IGF receptors IGF1R and IGF2R. The teeth were in different stages of resorption and all showed reparative cementum formation. It was found that acellular extrinsic fiber cementum, reversal lines and reparative cellular intrinsic fiber cementum were immunoreactive for both IGFs and various IGFBPs. Therefore, in human deciduous teeth, all subgroups of cementum, but not dentine, may represent sources of components of the IGF system. Odontoclasts did not carry IGFs or the IGF1R, but IGFBPs and the IGF2R. Therefore, these cells, in contrast to osteoclasts, may not respond to IGFs, but may be involved in the release and sequestration of IGFs from cementum during the resorption process. In contrast to odontoclasts, cementoblasts and periodontal ligament (PDL) fibroblasts carried IGF1R. The influence of the IGF system on the function of these cells with respect to periodontal matrix turnover and cementogenesis is discussed. On the behalf of the IGFBP immunoreactivities found, the PDL extracellular matrix can be considered to be a reservoir for IGF system components, where binding proteins may regulate IGF distribution and activity.     

Prospects for applying fluorescence spectroscopy to diagnose the hard tissues of a tooth

This article deals with the autofluorescence spectra from the hard tissues of a tooth, both in norm and pathology. An investigation was made on 30 extracted human teeth. The measurements were made both for the intact hard tissues of a tooth, such as enamel, dentine, cementum, and root canal, and for the tissues pathologically affected by a caries (superficial, intermediate, and deep) and by a dental calculus. It was found that the fluorescent spectra from enamel, dentine, cementum, and from the regions affected by a caries and dental calculus were identical in form. All the spectra revealed a maximum near 700 nm. However, the intact and affected hard tissues were greatly different in the integral fluorescent intensity. Dental calculus was found to produce the most pronounced fluorescent intensity, whereas the carious regions produced a slightly weaker fluorescent intensity. On the contrary, the intact hard tissues of a tooth exhibited the poorest fluorescent intensity.     

Inhibition of prostanoid synthesis depresses alveolar bone resorption but enhances root resorption in the rat

Tooth drift requires the deformation of the root socket and the adjustment of the other components of the attachment apparatus, namely, the periodontal ligament (PDL) and the cementum. Indomethacin (7.5 mg/kg/d), an inhibitor of prostanoid synthesis, provoked in rats a depression in the bone resorption effecting the deformation of the socket (Lasfargues and Saffar, Anat. Rec., 234:310–316, 1992). In the present paper we examined the consequence of this treatment both on the PDL and the root surface. After 3 days of treatment, when osteoclastic resorption was not yet disturbed, the root had been markedly resorbed (P<0.05) opposite the resorbing bone surface; at that time the PDL width remained in the normal range. After 7 days, i.e., when the bone resorption was depressed, the PDL was widened as the result of the ongoing root resorption. Despite the extensive root resorption, the anchorage of the PDL fibers appeared to remain effective, suggesting that it was rapidly restored. On day 14 at the time of the bone resorption recovery, cementum was deposited in the root resorption lacunae and the PDL width had returned to its control value. As early as day 3 the daily rate of dentine formation increased in the pulp area subjacent to the root resorption lacunae (P<0.01). These data demonstrate that (i) the responses of the different components of the periodontal apparatus are coordinated to allow for the maintainance of the PDL width so that when bone resorption is disturbed, root resorption compensates for it, and (ii) the odontoclasts can differentiate and resorb under prostanoid inhibition whilst osteoclastic resorption of the bone socket is inhibited. © 1993 Wiley-Liss, Inc.     

Histological study of deposited cementum in human deciduous teeth with pathological root resorption

Physiological root resorption is a characteristic feature of human deciduous teeth. Pathological root resorption due to apical periodontitis, dental trauma or excessive orthodontic force is also observed in deciduous roots. The root resorption is not continuous, and has resting periods. In the resting period, cementum deposits in resorbed root surface. The deposited cementum in permanent teeth has been reported in detail. However, the deposited cementum in deciduous teeth is unclear. The present study examined apices of roots of human deciduous incisors with apical periodontitis and roots of sound deciduous incisors by light and transmission electron microscopy. Root dentin and original cementum had a severe irregular caved surface. Cementum was partially deposited on the caved root surfaces. The deposited cementum had made the caved root surface relatively flat. The cementum was lax and had some defects. The deposited cementum was belt-like in shape and had a stratified structure. Each layer had various structures consisting of abundant microfibrils and fine granular materials, microfibrils, granular materials, and collagen fibrils, a few fibrils and granular materials and a relatively homogeneous structure. The original cementum had many collagen fibrils, such as intrinsic and extrinsic fibers, and no granular materials or homogeneous structure. Therefore, structure of the deposited cementum was very different from that of original cementum in deciduous teeth and from that of deposited cementum in permanent teeth.     

Scanning microscopy of platypus teeth

Summary Anorganic unerupted developing teeth and airdired erupted teeth of the platypus (Ornithorhynchus anatinus) were examined in a scanning electron microscope and in a tandem scanning reflected light microscope. Typically mammalian developing fronts of enamel and dentine were identified in the anorganic unerupted specimens. The developing teeth were particularly small and fragile and the enamel elusive and difficult to examine in the normal way for morphological detail. Prepared fractured surfaces of unerupted specimens revealed preferentially oriented crystallite groups in the enamel generally perpendicular to the developing front and a highly globular, mineralized pattern in the dentine with fine diameter, sparsely distributed dentinal tubules.Although optically homogeneous, the enamel of both developing and mature teeth displayed well-defined incremental lines, radial clefts, crystallite domains of variable size and outline, and fine tubules when examined by high contrast, back-scattered electron imaging. The enamel is prismatic only in part; well-formed, regular prisms not being a primary feature of platypus enamel. This can be related to the variability inherent in the developing surface and the thinness of the enamel layer. No surface was found which could be confidently identified as cementum; those developing surfaces not covered by enamel displaying small calcospherites which elsewhere marked the outer aspect of the dentine.     

Odontoclastic resorption at the pulpal surface of coronal dentin prior to the shedding of human deciduous teeth.

Histological and histochemical observations of more than 150 extracted human deciduous teeth revealed that, prior to shedding, odontoclastic resorption as a rule takes place at the pulpal surface of coronal dentin. We also found that this phenomenon occurs in all kinds of deciduous teeth. The process of this internal resorption of coronal dentin of deciduous teeth clearly showed time-related histological changes. During the time the roots were actively being resorbed, the pulpal tissue retained its normal structure. However, when root resorption neared completion, inflammatory cells started to gradually infiltrate into the pulp, and odontoblasts began to degenerate. After that, multinucleate odontoclasts appeared, and resorption proceeded from the predentin to the dentin. The odontoclastic activity was initially detected only on the pulpal surface at the bottom areas of the crown. It gradually spread towards the pulpal horn regions along the wall of the pulp chamber. However, this internal resorption of coronal dentin did not continue until the teeth were finally shed. After the elimination of resorption, the resorbed dentin surface was repaired by a cementum-like deposition or covered with fibrous connective tissue.     

The odontoclasts of Ambystoma mexicanum

The resorption of teeth in Ambystoma mexicanum during postembryonal ontogenesis and induced metamorphosis occurs by means of light-microscopic detectable giant-cells. These have morphological and functional characters similar to those of odontoclasts of other vertebrates. The multinucleated odontoclasts resorb not only the pedicel (base), but the stalk of the tooth, too. When active, the cells form a ruffled border and a sealing zone. In this way they are able to demineralize the hard tissues of teeth (dentin and mineral of the pedicel) and to dissolve the extracellular matrix. Resorption of enamel has not been observed. Marks of resorption resemble the Howship's lacunae of other tetrapods. TRAP as a typical enzyme of odontoclasts could not be detected histochemically. Dependence of PTH, which is supposed to be necessary for the formation and activation of odontoclasts as well as of thyroxine can be excluded, although the resorbing cells are functionally and cytologically identical with those of other vertebrates. This demands some other mechanism for the formation and regulation of the odontoclasts in A. mexicanum.     

Features of the clear zone of odontoclasts in the Chinook salmon (Oncorhynchus tshawytscha)

This study aims to clarify the features of the clear zone of odontoclasts on shedding teeth of a teleost fish, Chinook salmon, Oncorhynchus tshawytscha (Walbaum), using a light microscope to determine the orientation between a cell body and a resorptive lacuna, followed by transmission electron microscopy. Ultrathin sections of LR White embedded material were incubated in rabbit anti-actin polyclonal antibody and then were incubated with 15 nm gold-conjugated goat anti-rabbit IgG. The clear zones of odontoclasts showed a variable structure with electron-dense structures on sections, but distinct clear zones were not always seen on odontoclasts. In odontoclasts sectioned in the direction perpendicularly to the surface of a resorptive lacuna, some cells showed a wide clear zone, but two types of clear zones were usually observed: a part composed of some cytoplasmic processes and one composed of several complicatedly interwoven processes. Gold particles were localized on the clear zones, especially in electron-dense structures; very few gold particles were detected in ruffled borders. These results show that the clear zone of odontoclasts in Chinook salmon contains actin. Our results suggest that the clear zone of an odontoclast in Chinook salmon is not always a wide annular structure.     

Mononuclear odontoclast participation in tooth resorption: The distribution of nuclei in human odontoclasts

Osteoclasts and odontoclasts have been considered multinucleated giant cells which resorb hard tissue by ruffled borders. Recently, the authors reported the presence of a mononuclear osteoclast and odontoclast with a ruffled border. However, the relative frequency of such cells and the distribution of the number of nuclei including mononuclear cells in them have not been elucidated. Six human deciduous teeth were used in this study. After fixation and decalcification, tartrate-resistant acid phosphatase (TRAP) activity was detected with the azo dye method, and then TRAP-positive cells were observed on resorbing areas of teeth by light microscopy. The cells for investigation were serially sectioned by semithin sections to observe the presence of resorptive lacuna and the number of nuclei. The TRAP activity was detected in both multinucleated and mononuclear odontoclasts from serial semithin sections, and 242 TRAP-positive cells which formed lacunae on dentin were investigated to determine the frequency distribution of the number of nuclei. The mean number of nuclei per cell was 5.3, and median was 4. Only 2.9% of odontoclasts were mononucleus and 93.8% had 10 or fewer nuclei. The majority of odontoclasts forming lacunae on the dentin were cells with 10 or fewer nuclei, and mononuclear odontoclasts participated in human deciduous tooth resorption together with multinucleated ones. Anat. Rec. 249:449–457, 1997. © 1997 Wiley-Liss, Inc.