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ABSTRACT: The three-dimensional architecture of enamel prisms at early stages of enamel formation and its spatial relationship to the Hunter-Schreger bands were examined in canine tooth germs by light and electron microscopy. In serial semithin sections of demineralized tooth germs tangential to the enamel-dentin junction, a straight row of enamel prisms was depicted along the longitudinal tooth axis at the level of the enamel-dentin junction and then their three-dimensional arrangement was reconstructed using computer software. The spatial arrangement of the groups of enamel rods oriented in specific sideward directions was also reconstructed in deep layers of the enamel. Initially, all enamel prisms were parallel to perpendicular toward the enamel-dentin junction, but at 10µm from the enamel-dentin junction, some small specks, or groups of enamel prisms--tilting to the right or the left--emerged as small islands. In each speck of enamel prism, the inclined prisms were uniformly oriented in a sideward direction and gradually expanded their boundary until merging with the neighboring specks inclined in the same direction. Consequently, at 50µm from the enamel-dentin junction, the group of enamel prisms oriented either to the right or the left formed alternately arranged horizontal belt-like zones, corresponding to the parazone or the diazone of the Hunter-Schreger bands. Reversed images of scanning electron-micrographs of the exposed surfaces of the developing enamel revealed round and bulb-like profiles of Tomes' processes at early amelogenesis and its changes into a characteristic structure combined with flat secretory and enclosing nonsecretory faces that dictated the orientation of corresponding enamel prisms. The results suggest that the groups of enamel prisms oriented in sideward directions first appear as small island-like specks near the enamel-dentin junction, which later merge and form alternating horizontal belt-like zones as a consequence of morphological changes of the Tomes' processes. However, the mechanisms whereby the functional grouping of secretory ameloblasts with similarly oriented Tomes' processes is induced are yet to be determined.
Archives of Histology and Cytology 01/2010; 73(1):23-36. · 0.57 Impact Factor
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ABSTRACT: In submandibular gland atrophy, most acinar cells disappear by apoptosis, while many duct cells remain. The present study aimed to establish whether Bcl-2 and Bax, members of the Bcl-2 gene family, regulating the signalling pathway of apoptosis were involved in duct cell survival and acinar cell death in atrophic submandibular glands. The excretory duct of rat submandibular gland was doubly ligated with metal clips from 1 to 14 days to induce atrophy to the gland. The expressions of Bcl-2 and Bax in the atrophic submandibular gland were examined using immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR). Immunohistochemically, Bcl-2 expression was identified in duct cells in the experimental glands at all time points. Some acinar cells showed Bax positivity 1 day after excretory duct ligation, and there were more Bax-positive acinar cells on days 3 and 5 when many apoptotic acinar cells were observed. Analysis by RT-PCR showed that the expression of mRNA for Bcl-2 became stronger as the glandular atrophy progressed and that Bax mRNA strongly expressed on days 1 and 3. These observations suggest that Bcl-2 inhibits duct cell apoptosis and Bax promotes apoptosis of acinar cells during atrophy of submandibular glands.
International Journal of Experimental Pathology 11/2008; 89(5):303-8. · 2.57 Impact Factor
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ABSTRACT: This study was designed to establish the apoptosis of odontoclasts during physiological root resorption of human deciduous teeth. Deciduous teeth were fixed, decalcified, and embedded in paraffin for immunohistochemical (IHC) observations and in Epon for transmission electron microscopy (TEM). Apoptotic cells were identified by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick-end labeling (TUNEL), and then tartrate-resistant acid phosphatase (TRAP) activity was determined on the same sections. Epon-embedded specimens were sectioned serially into 0.5-microm semithin sections; some of these sections were re-embedded in Epon, sectioned into 0.1-microm ultrathin sections, and observed by TEM. IHC revealed that the nuclei of TRAP-positive odontoclasts on the dentine were generally TUNEL-negative. Around these odontoclasts, a few TRAP-positive structures were present together with TUNEL-positive structures, e.g., a TRAP-positive structure with one TUNEL-positive nucleus, a TRAP-positive structure with one TUNEL-positive nucleus plus one or two TUNEL-negative nuclei, or a TRAP-positive structure with no nucleus. By TEM, some odontoclasts showed nuclear fragments including compacted chromatin. The results suggest that, during apoptosis, odontoclasts fragment into variously sized cellular parts including three or fewer nuclei.
Cell and Tissue Research 03/2008; 331(2):423-33. · 3.11 Impact Factor
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ABSTRACT: This study was designed to detect tissue non-specific alkaline phosphatase (TNSALP) by Azo-dye staining, calcium by glyoxal bis (2-hydroxyanil) (GBHA) staining, bone sialoprotein (BSP) and osteopontin (OPN) by immunoperoxidase staining in developing rat molars, and also to discuss the mineralization process during acellular cementogenesis. To restrain a reduction in histochemical and immunohistochemical reactions, fresh-frozen undemineralized sections were prepared. Where the epithelial sheath was intact, TNSALP reaction was observed in the dental follicle, but not in the epithelial sheath. With the onset of dentin mineralization, the BSP- and OPN-immunoreactive, initial cementum layer appeared. At this point, cementoblasts had shown intense TNSALP reaction and GBHA reactive particles (=calcium-GBHA complex) appeared on the root surface. With further development, the reaction of TNSALP and GBHA became weak on the root surface. Previous studies have shown that the initial cementum is fibril-poor and that matrix vesicles and calciferous spherules appear on the root surface only during the initial cementogenesis. The findings mentioned above suggest that: during the initial cementogenesis, cementoblasts release matrix vesicles which result in calciferous spherules, corresponding to the GBHA reactive particles. The calciferous spherules trigger the mineralization of the initial cementum. After principal fiber attachment, mineralization advances along collagen fibrils without matrix vesicles.
Histochemie 04/2007; 127(3):303-11. · 2.59 Impact Factor
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ABSTRACT: Most acinar cells and some duct cells undergo apoptosis during atrophy of the submandibular gland. The present study was designed to elucidate whether Fas and its receptor ligand (FasL) are involved during apoptotic atrophy of the gland. The excretory duct of the right submandibular gland of rats was doubly ligated with metal clips from 1 to 14 days for induction of gland atrophy. Control rats were untreated. Fas and FasL expression in the atrophied submandibular gland was detected using immunohistochemistry (IHC) and Western immunoblot. Expression of activated caspase 8 and activated caspase 3 was also detected with IHC. Fas-positive acinar and duct cells and FasL-positive duct cells increased in the atrophic glands at 3 and 5 days after duct ligation when apoptotic cells were commonly observed. Thereafter, Fas- and FasL-positive cells declined in number. Patterns of expression of Fas and FasL using Western immunoblots concurred with the IHC results. Activated caspase 8-positive cells were present at every time interval but peaked at 3 and 5 days following duct ligation. The cells showing immunoreaction for activated caspase 3 first appeared on day 3, with the peak in apoptosis, after which they decreased. The results indicate that the Fas/FasL systems likely play an important role in apoptotic pathways during atrophy of the submandibular gland.
International Journal of Experimental Pathology 03/2007; 88(1):9-17. · 2.57 Impact Factor
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ABSTRACT: 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.
Anatomy and Embryology 04/2006; 211(2):87-93. · 1.42 Impact Factor
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ABSTRACT: Odontoclasts resorbing teeth are multinucleated cells. Previously, the authors have investigated the distribution of number of nuclei per human odontoclast and showed that the mean number of nuclei per cell is 5.3, the median is 4, and 93.8% of cells have 10 or fewer nuclei. Teleost odontoclasts have features similar to those of mammals; however, the distribution of number of nuclei per cell remains unknown. The present study aimed to examine the distribution of number of nuclei per odontoclast in a teleost fish, Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and to clarify the difference of number of nuclei in odontoclasts between Chinook salmon and humans. The maxillae and mandibles of Chinook salmon were fixed, decalcified, and embedded in Epon 812. Specimens were serially sectioned into 0.5-microm semithin sections and examined by light microscopy. Cells possessing a brush border adjacent to a resorptive lacuna were identified as odontoclasts, and 246 odontoclasts were investigated to determine the distribution of nuclei per cell. The mean number of nuclei per cell was 21.8 and the median was 17; only 24.4% of odontoclasts had 10 or fewer nuclei, and 95.5% had 50 or fewer nuclei. These results suggest that the range for the number of nuclei per odontoclast in Chinook salmon is greater than that in humans.
Anatomy and Embryology 01/2005; 209(2):119-28. · 1.42 Impact Factor
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Clinical calcium 02/2004; 14(1):195-8.
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ABSTRACT: Many biochemical reports support cell-cell interaction between osteoclasts and osteoblasts/stroma cells in vitro, however there have been few morphological studies supporting this. Details of cell-cell interaction between osteoclasts and osteoblasts/stroma cells remain unclear. The present study examined cell-cell interaction between osteoclasts and osteoblasts/stroma cells by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Osteoclasts, osteoblasts/stroma cells, and bone marrow cells obtained from 10-day-old ddY mice were cultured on dentin slices for 72 hr. Specimens were fixed, and some were examined by SEM. Specimens were decalcified, embedded in Epon after determination of the tartrate-resistant acid phosphatase activity (TRAP), and TRAP-positive cells for investigation were serially sectioned by alternating semithin and ultrathin sections, and then examined by TEM. By SEM, many cellular contacts were seen between the cells cultured on the dentin, but by TEM there were few special structures on the cell membranes between osteoclasts and osteoblasts/stroma cells, or between osteoclasts and bone marrow cells. A special structure on the cell membranes of osteoclasts was observed between an osteoclast and a cytoplasmic process of osteoblast/stroma cells, and this cell membrane was coated with electron dense or bristle-like structures. These bristle-like structures were very similar to those of coated pits. The present results show that the coated pit-like structure plays an important role in cell-cell interaction between osteoclasts and osteoblasts/stroma cells in vitro, and suggest that macromolecules binding to the osteoclast-surface receptor via ligands, accumulate in the coated pits, and enter the osteoclast as receptor-macromolecule complexes in endocytic vesicles.
Annals of Anatomy - Anatomischer Anzeiger 06/2002; 184(3):221-7. · 1.86 Impact Factor
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ABSTRACT: The cemento-dentinal junction was observed in the acellular and cellular cementum of rat molars by light and scanning electron microscopy. Scanning electron microscopy, combined with NaOH maceration, was used to observe the fibrous architecture directly in this region. Light microscopy revealed that the cemento-dentinal junction contains fewer collagen fibrils and more proteoglycans than the cementum and dentin. Scanning electron microscopy also showed that fibril intermingling is found only in some regions of the fibril-poor junction in macerated specimens. Prolonged maceration breaks down the cemento-dentinal junction in spite of the fibril intermingling. Only macerated specimens showed detachment here. It was established that NaOH maceration removes interfibrillar substances effectively, and does not damage the fibril structure or architecture. This suggests that the adhesion of proteoglycans is more important than fibril intermingling for preserving the cemento-dentinal attachment in the rat molar.
Annals of Anatomy - Anatomischer Anzeiger 04/2000; · 1.86 Impact Factor
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ABSTRACT: The distribution of the myoepithelial cells during regeneration of the rat parotid gland after atrophy induced by one week of parotid duct ligation was investigated by immunohistochemistry for actin and transmission electron microscopy (TEM). Immunohistochemically, residual ducts were surrounded by actin-positive cells when clips were removed from the duct. Three days later, most of the newly formed acini originating from the residual ducts were also embraced by actin-positive cells. After 10 days, actin-positivity tended to be seen as dots around acini that decreased in number day by day. On day 21 actin-positive cells mainly surrounded intercalated ducts with only a few positive reactions identified at the acinar periphery. Electron microscopically, residual ducts and newly formed acini were peripherally embraced by myoepithelial cells before day 5. After day 7, shift of myoepithelial cells from the periphery of acini to the duct-acinar junctional region was identified. Then few myoepithelial cells were identified at the periphery of acini. These observations indicate that myoepithelial cells migrate from the acinar periphery to the duct-acinar junctional region during rat parotid regeneration, and that such behaviour is closely related to that seen during rat parotid development.
International Journal of Experimental Pathology 09/1999; 80(5):283 - 290. · 2.57 Impact Factor
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ABSTRACT: The formation of intrinsic fibers was examined in the advanced stage of rat cellular cementogenesis by light and electron microscopy. Using scanning electron microscopy, cementoblasts showed wing-like processes, partly encircling principal fibers. At the cementum-facing side of the cells these processes showed segmentation into finger-like processes, arranged in parallel with the cementum surface. Transmission electron microscopy showed many cytoplasmic fragments around intrinsic fibers at the cementum surface. These fragments contained microtubules and collagenous secretory granules that were arranged in parallel with the cementum surface and the intrinsic fibers. The wing-like processes contained microtubules and secretory granules that were arranged perpendicularly to the cementum surface and in parallel with the principal fibers. These observations suggest that: (1) the cytoplasmic fragments are cross-sectioned finger-like processes; (2) cementoblasts secrete intrinsic fibers from the finger-like processes and additional principal fibers from the wing-like processes; (3) cementoblasts constantly shorten their wing-like processes by forming finger-like processes. This development starts at the side facing the cementum and proceeds towards the periodontal ligament. With the segmentation, the cementoblasts change the arrangement of secretory granules to secrete intrinsic fibers around preformed principal fibers.
Anatomy and Embryology 04/1996; 193(5):495-500. · 1.42 Impact Factor
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ABSTRACT: The initial genesis of acellular and cellular cementum was examined in rat molars by light and electron microscopy. Before root dentinogenesis, flattened dental follicle cells formed compartments by regularly arranged cellular processes which demarcated collagen fibril bundles oriented in parallel with the root long axis in both of the two kinds of cementum. After this stage, compartments disappeared from the dental follicle cells, which became elongated and polarized, with the cytoplasmic side facing toward the root surface in the acellular cementogenesis. Fibril bundles, oriented in parallel with the root long axis, decreased in number, and principal fibers appeared. Some principal fibers were attached on the first acellular cementum. Observations suggested that the fibril bundles, which had been oriented in parallel with the root long axis, were reoriented to merge into the principal fibers. In cellular cementogenesis, the dental follicle cells continued to hold the fibril bundles in cellular compartments. The regular processes were transformed into randomly oriented, finger-like processes. At the same time, fibers, which may be secreted from the finger-like processes, appeared around the preformed fibril bundles oriented in parallel with the root long axis. The different cellular behavior may result in the different fiber arrangement of acellular and cellular cementum.
Anatomy and Embryology 01/1994; 190(6):521-527. · 1.42 Impact Factor
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ABSTRACT: The initial attachment of principal fibers to the root dentin surface was investigated in developing rat molars by light and electron microscopy. Ruthenium red was used to intensely stain the first deposited material on the root dentin surface. Observations suggested that when the principal fibers became embedded in the ruthenium red-positive material, there was no complicated interweaving between the principal fibers and the root dentinal matrix fibers. Therefore, it is likely that the initial principal fiber attachment to the root dentinal matrix fibers is mediated by the ruthenium red-positive material.
Journal of Periodontal Research 02/1990; 25(2):113 - 119. · 1.69 Impact Factor
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ABSTRACT: In a reversal phase of bone remodeling many mononuclear cells appear on the resorbed surfaces of bone with characteristic reversal lines as revealed by transmission electron microscopy (TEM). However, these mononuclear cells have been variously hypothesized or reported. The present study examined the TEM features on the resorbed surfaces of three calcified connective tissues, and aimed to clarify the nature and function of the mononuclear cells in a reversal phase. Dentine slices cultured with isolated osteoclasts, human deciduous teeth, and rat mandibles were used in this study. Specimens were fixed, decalcified, and then embedded in Epon 812, and sectioned into 0.1-μm-thick ultrathin sections. The ultrathin sections were stained with uranyl acetate and lead citrate, and then examined by TEM. Many sharply pointed collagen fibrils with striation were observed exposed on the resorbed surfaces of cultured dentine slices, but there were neither cells nor reversal lines. The same features were observed on the root dentine surfaces of human deciduous teeth. Under many mononuclear cells in a reversal phase of remodeling, reversal lines were seen on the resorbed surfaces of rat mandibles, but there were no striated collagen fibrils exposed on the bone surfaces. The alternation of the TEM features on the resorbed surfaces before and after the participation of mononuclear cells in a reversal phase of remodeling suggests the nature and function of these cells: they participate in both degrading the demineralized and disrupted matrix left on the resorbed surfaces and forming reversal lines there.
Annals of Anatomy - Anatomischer Anzeiger.
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ABSTRACT: This study was designed to observe drifting molars of 70-day-old rats by light microscopy, and to elucidate whether there are similar attachment mechanisms at different kinds of adhesive lines in periodontal mineralized tissue of the rat molar region. Three kinds of adhesive lines — cement lines on resorbed alveolar bone, cement lines on resorbed roots, and cemento-dentinal junctions were examined. The two kinds of cement lines showed similar histological and histochemical features, they were proteoglycan-rich and fiber-poor. They appeared to form on the resorbed tissue before principal fiber reattachment. After covering by new bone or by reparative cementum, the cement lines retained the original features. The cemento-dentinal junction showed features very similar to those of the cement lines. Previous studies have suggested that the cemento-dentinal junctions bind the cementum and dentine by adhesion of proteoglycans. Structural similarities suggest that cement lines provide similar links between new bone and resorbed bone and between resorbed root and reparative cementum. In conclusion, this study suggests that there is one attachment mechanism for the different kinds of collagen based hard tissue in the rat molar region.
Annals of Anatomy - Anatomischer Anzeiger.
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ABSTRACT: 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 μm 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.
Annals of Anatomy - Anatomischer Anzeiger.
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ABSTRACT: Resin-embedded transparent teeth are devoid of the original crown morphology, because enamel is lost by demineralization. This study was designed to reproduce artificial enamel and to reconstruct the original crown morphology for resin-embedded transparent teeth.
The impression of the coronal portion of human permanent teeth was taken with a silicone impression material. After demineralization, drawing ink was injected into the pulp cavities. The ink-infiltrated teeth were made transparent with methyl salicylate and embedded with polyester resin. Urethane prepolymer was injected into the impression, and the resin-embedded teeth were reinserted into the impression. After polymerization of the urethane resin, the specimens and the urethane resin were removed from the impression.
The original crown morphology of the resin-embedded transparent teeth could be precisely reconstructed with artificial and removable enamel. The resin-embedded teeth showed morphologic details of the black-stained pulp cavities through the transparent dentin and cementum.
This study established a crown reconstructing method for resin-embedded transparent teeth. The specimens will be useful for demonstration of morphology of teeth and pulp cavities.
Quintessence international 34(10):749-51. · 0.76 Impact Factor
