Noriyuki Sahara

Cytodifferentiation of the Odontoclast Prior to the Shedding of Human Deciduous Teeth: An Ultrastructural and Cytochemical Study

In human deciduous teeth, odontoclastic resorption takes place at the pulpal surface of the coronal dentine prior to shedding, and this resorption shows clear time‐related histological changes (Sahara et al., 1992).

Effect of streptozotocin-induced diabetes mellitus on alveolar bone deposition in the rat

Effects of diabetes on alveolar bone remodelling were assessed by quantitative histology and a chronological lead-labelling technique. Experimental diabetes was induced by a single dose of 40 mg/kg of streptozotocin. Remodelling of the alveolar wall surrounding the root of mandibular first molar was studied in control rats fed ad libitum, and in diabetic and insulin-treated diabetic rats 24 days after the induction of diabetes. The volumes of bone formation on the mesial side of the alveolar wall were evaluated over a 10-day period by chronological lead-labelling and computer image analysis. For a histometric measure of bone-resorption, the number of osteoclasts along the distal surface of the alveolar wall was counted. The volume of bone formed and the number of osteoclasts were significantly lower in the diabetic rats than in the controls, but insulin treatment of diabetic rats normalised these histomorphometric measures of bone turnover. These results demonstrate that streptozotocin-induced diabetes mellitus reduces the rate of bone turnover in the alveolar wall surrounding the root, which reduction is corrected by treatment with insulin.

Odontoclastic resorption of the superficial nonmineralized layer of predentine in the shedding of human deciduous teeth

Resorption by odontoclasts of a superficial nonmineralized layer of predentine that occurs in prior to the shedding of human deciduous teeth was studied by light and electron microscopy. As resorption of the tooth roots neared completion, multinucleate cells appeared on the predentine surface of the coronal dentine between the degenerated odontoblasts, excavated characteristic resorption lacunae in the nonmineralized predentine. These multinucleate cells had the same ultrastructural characteristics as odontoclasts and histochemical demonstration of tartrate-resistant acid phosphatase activity in the multinucleate cells revealed intense staining in numerous small granules identified as lysosomes. Occasionally, the multinucleate cells simultaneously resorbed both nonmineralized and calcospherite-mineralized matrix in the predentine. The study demonstrates that multinucleate odontoclasts can resorb nonmineralized predentine matrix in vivo, probably in the same way as they resorb demineralized organic matrix in the resorption zone underlying their ruffled border.

Temporal changes in the distribution and number of macrophage-lineage cells in the periodontal membrane of the rat molar in response to experimental tooth movement

In order to evaluate the possible role of macrophages in the remodelling of periodontal tissue in response to tooth movement, temporal changes in the number and distribution of macrophage-lineage cells in the periodontal membrane of the rat molar tooth after experimental tooth movement were examined immunohistochemically using four anti-rat monoclonal antibodies: ED1 (anti-monocyte/macrophage-lineage cells and dendritic cells), ED2 (anti-resident macrophages), KI-M2R (anti-tissue macrophages), and OX6 (anti-class II molecules). The right maxillary first molar tooth of Wistar rats was moved mesially by a closed-coil spring for 1, 3, 5, or 7 days. Sham-treated rats wearing an inactivated appliance for each experimental period and entirely untreated rats were used as controls. Alternate horizontal serial cryostat sections were cut and incubated with antibodies to ED1, ED2, KI-M2R, and OX6. In addition, cells immunopositive for each monoclonal antibody in the periodontal membrane during tooth movement were analysed on the tension and pressure sides. In the control rats, large numbers of cells positively stained with each monoclonal antibody were distributed throughout the periodontal membrane surrounding the distobuccal root. At 1 day after experimental tooth movement, the number of immunopositive cells obtained with all four monoclonal antibodies decreased as compared with those of the control on the mesial/pressure side. During the later experimental time periods, ED1- and OX6-positive cells in the periodontal membrane of this side were significantly increased in number compared with controls, whereas the density and distribution pattern of cells positive with ED2 or KI-M2R remained unchanged. On the mesial/pressure side, which underwent hyalinization, a marked accumulation of OX6- and ED1-reactive cells, but not of ED2- or KI-M2R-reactive cells, was frequently observed in the area of the hyalinized tissue at 5-7 days after the start of tooth movement. On the distal/tension side, no particular change in the distribution of immunopositive cells obtained with any antibody was detected throughout the experimental periods, with the exception that there was a significant increase in the number of ED1-positive cells and in of OX6-positive cells at 1 and 7 days, respectively, after the start of tooth movement. These results suggest that after the start of tooth movement OX6- and ED1-positive cells, which are mostly exudative macrophages, but not ED2- and KI-M2R-positive cells, i.e., resident macrophages, may be actively engaged in bone resorption and the remodelling of tissues on the pressure side of the periodontal membrane.

Hard Tissue Formation in Subcutaneously Transplanted Rat Dental Pulp

While dental pulp appears to be able to form mineralized matrices that do not always resemble dentin, the precise characteristics of the hard tissue and the mechanism of its induction remain unknown. Therefore, we evaluated hard tissue induced by transplantation of pulp into subcutaneous tissue. Seven days after transplantation, initial hard tissue was formed at the inner periphery of the pulp. After 14 days, this hard tissue expanded inwardly. Mineralized matrix was immunopositive for osteocalcin, osteopontin, and bone sialoprotein, but negative for dentin sialoprotein. Transplantation of GFP-labeled pulp into wild-type rats showed these formative cells to have been derived from the transplant. TEM observation revealed apatite crystals within necrotic cells and matrix vesicles at the initial stage of calcification. These results indicate that pulp cells possess the ability to form a bone- or cementum-like matrix. Calcification of the matrix may occur in necrotic cells and matrix vesicles, followed by collagenous calcification.

Quantitative evaluation of newly formed bone in the alveolar wall surrounding the root during the initial stage of experimental tooth movement in the rat

By using a chronological lead-labelling technique and computer image analysis the volume of this newly formed bone was evaluated. Rat maxillary first molars were moved mesially by a fixed, closed coil-spring appliance for 6 days using three different magnitudes of initial tensile force (27, 60 and 136 g). Sham-treated rats wearing an inactivated appliance were used for the control study. All animals were injected twice intraperitoneally with lead-disodium EDTA, 3 hr before the beginning and 3 hr before the end of treatment. The unit volumes of newly formed bone (mm3/mm2) were assessed with reference to lead-labelling lines in the alveolar walls of the root socket by computer image analysis. In the control group, two distinct lead-labelling lines indicated continuous bone formation on the mesial side of the root sockets, but only a jagged line was found on the distal side. After experimental mesial tooth movement, only a single lead line could be found on the mesial/pressure side of the root sockets; on the distal/tension side, a wide layer could be detected between the two lead lines. The volume of newly formed bone on the distal/tension side in the experimental groups was significantly greater than that in the control group. However, there was no significant difference in the volumes of newly formed bone among the three experimental groups. The study demonstrates that the volume of newly formed bone in the alveolar walls during the initial stage of tooth movement can be quantified and that the magnitude of the tensile force of tooth movement may not influence directly the volume of newly formed bone in the alveolar wall on the tension side.

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.

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.

A Histological Study of the Exfoliation of Human Deciduous Teeth

For clarification of the histological details of the shedding of human deciduous teeth, exfoliated and extracted deciduous teeth were examined by light and electron microscopy. After the roots were completely resorbed, the dento-gingival junction migrated along the inner resorbing surface and finally reached the pulpal surface of the crown. At the same time, the gingival epithelium also proliferated and migrated under the crown of the deciduous tooth in such a way that part of it lined the residue of the pulp and another part lined the surface overlying the erupting successional tooth. This phenomenon took place from various sides of the tooth surface. Therefore, just before exfoliation, the migrated gingival epithelium formed narrow necks of tissue, and the crown was only superficially attached to the gingiva by them. The final shedding of the tooth appeared to occur by a tearing of these narrow tissue regions. The results of the present study suggest that the dento-gingival junction as well as gingival epithelium play important roles in the process of exfoliation of human deciduous teeth.

Fine structure of submandibular glands of mice with testicular feminization (Tfm/Y)

SummaryThe fine structure of the submandibular gland of the mouse with testicular feminization (Tfm/Y) was studied by light and electron microscopy. The architecture of the Tfm/Y gland proved to be rather similar to that of the normal female mouse in both tubular ratio and structure. Granular convoluted tubular cells in Tfm/Y mice characteristically had fewer secretory granules and increased cytoplasmic vacuoles than normal littermates, suggesting an altered synthesis of secretory granules in this cell type of the Tfm/Y mouse. Moreover, there were differences in the ultrastructure of submandibular glands between Tfm/Y and normal female mice. In the gland of the Tfm/Y mouse, basal striations of the striated secretory tubular cells were not so developed and granular intercalated duct cells were less than those of normal females. These findings support the evidence that the secretory tubule of the mouse submandibular gland responds to androgens, resulting in accentuated development in the male, while also suggesting the possibility that the mouse submandibular gland is regulated by other factors which lead to the prominent sexual dimorphism observed in this gland.