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Y Sasano

Publications and source records attributed to Y Sasano.

At least 37 records · Page 2Linked to original sources

[Cementum formation in rat molar roots].

Cementum is the calcified tissue covering roots of teeth and serves as attachment sites of the periodontal ligament. Although recent studies have suggested that extracellular matrix of cementum is very similar to that of bone, cementogenesis on a biological basis is still poorly understood. There are variations in the distribution and mineral contents of cementum depending on animal age, tooth species and position within the tooth roots. This paper reviews the formation and age-related changes of cellular and acellular cementum in rat molar roots, and discusses the effect of mechanical stress to the cementum formation.

Animals↗

Implanted octacalcium phosphate (OCP) stimulates osteogenesis by osteoblastic cells and/or committed osteoprogenitors in rat calvarial periosteum.

Our previous studies demonstrated that the octacalcium phosphate (OCP) causes new appositional bone formation on the OCP when implanted into the subperiosteal region of murine calvaria. The OCP may stimulate the cell population committed to the osteoblastic differentiation in the periosteum and have them express the phenotype. The present study was designed to investigate which periosteal cell population is involved in bone formation on the OCP with applying the OCP implants on top of and underneath the periosteum. The periosteum of the rat parietal bones was flapped and the OCP was implanted on top of or underneath the periosteum, in which the implantation sites were defined using the membrane filter. The histology was examined to see if new appositional bone formation occurs on the OCP implant under each condition. New bone was deposited on the OCP on the bone surface separated from the periosteum by the filter, whereas no bone was formed either under the periosteum separated from the bone surface by the filter or on the periosteum. The present study suggests that the OCP acts on osteoblasts, bone lining cells and/or their closely committed progenitors on the bone surface to express the phenotype and deposit new bone on the OCP implant.

Animals↗

Confocal microscopy of dentinal tubules in human tooth stained with alizarin red.

The present study was designed to analyze the structures of dentinal tubules by confocal microscopy. Undecalcified ground sections of human teeth were stained with alizarin red in 0.1% KOH aqueous solution, and examined by confocal microscopy. Alizarin red stained dentinal tubules, interglobular dentine, granular layer of Tomes, and the surface of dentine. Interglobular dentine was seen between the outer and middle layers of coronal dentine. At the outer layer of coronal dentine, the dentinal tubules were thin and showed numerous branches. At the middle layer of coronal dentine, dentinal tubules displayed two types. The type I tubules are the dentinal tubules that do not show any nodular structures and the type II tubules are the dentinal tubules that appear bamboo-like with many nodules. In the cross section through the type II tubules, the nodules appeared as fine circular tubules surrounding the dentinal tubules. The circular tubules of nodules adhered to one side of the dentinal tubules. When the fluorescence images were compared with the images taken by transmission light mode, the fluorescence of dentinal tubules was seen at the inner surface of dentinal tubules, and the fluorescence of nodules was seen at interface between peritubular and intertubular dentine. Most of the dentinal tubules were of the type II tubules in the teeth from older individuals, whereas the type II tubules were scarce in the teeth from younger individuals. At the inner layer of coronal dentine, the dentinal tubules have no nodules and branches were scarce. The dentinal tubules of radicular dentine were different from those of coronal dentine. Most of the dentinal tubules were the type I tubules. Numerous fine branches were seen at the outer and middle layers of radicular dentine. No interglobular dentine was seen in the root except at the cervical part, and the granular layer of Tomes was also positive with alizarin red. At the cervical part of the root, interglobular dentine was present and the dentinal tubules displayed types I and II.

Adult↗

Implantation of octacalcium phosphate (OCP) in rat skull defects enhances bone repair.

Synthetic octacalcium phosphate (OCP) enhances bone formation if implanted into the subperiosteal region of murine bone. Such implanted OCP may be resorbed and replaced by bone with time. We hypothesized that OCP could be used as an effective bone substitute. To test this hypothesis, we designed the present study to investigate if bone repair in a rat skull defect is enhanced by the implantation of OCP. Rats were divided into two groups: OCP-treated animals and untreated controls. Six rats from each group were fixed at 4, 12, and 24 weeks after implantation. A full-thickness standardized trephine defect was made in the parietal bone, and synthetic OCP was implanted into the defect. After being examined radiographically, the specimens were decalcified and processed for histology. OCP implantation significantly promoted bone repair compared with the controls. A statistical analysis showed an increase in the area of radiopacity within the skull defect between week 4 and week 12. Histologically, bone was formed on the implanted OCP and along the defect margin at week 4. At week 12, the implanted OCP was surrounded by newly formed bone. At week 24, the defect was almost completely filled with bone. In the control, bone formation was observed only along the defect margin. The present results demonstrate that OCP could be used as an effective bone substitute.

Animals↗

Type X collagen is not localized in hypertrophic or calcified cartilage in the developing rat trachea.

Our previous studies have shown that rat tracheal chondrocytes become larger and hypertrophic, and that the cartilage matrix calcifies during development. Type X collagen is a short collagen molecule identified in hypertrophic and calcified cartilage in the growth plate of long bones during endochondral ossification. The present study was designed to investigate the distribution of type X collagen in rat tracheal cartilage during development before and after hypertrophization and calcification. Tracheas from postnatal Wistar rats, newborn, and at 4, 8 and 10 weeks were fixed along with hind limbs from newborn rats. Serial sections were made and adjacent sections were processed for von Kossa staining or immunohistochemistry for type X collagen. In addition, the immunoreactivity to type II collagen was examined as a control. The anti-type X collagen antibody stained hypertrophic and/or calcified cartilage in the newborn rat tibia. The immunoreaction for type X collagen was localized in the uncalcified peripheral region of tracheal cartilage in 4, 8 and 10-week-old rats. In contrast, the anti-type X collagen antibody did not show immunoreactivity to hypertrophic or calcified cartilage in the central region of the 10-week-old rat tracheal cartilage. The present study has suggested that type X collagen is not involved in hypertrophization of chondrocytes or calcification of the matrix in developing rat tracheal cartilage.

Animals↗

An immunohistochemical study of the localization of biglycan, decorin and large chondroitin-sulphate proteoglycan in adult rat temporomandibular joint disc.

To analyse regional variations in extracellular matrix components of adult rat temporomandibular joint discs, immunohistochemical techniques were used to examine the localization of two small dermatan-sulphate proteoglycans, biglycan and decorin, and a large chondroitin-sulphate proteoglycan. Staining for biglycan was intense in the posterior band, although it had a rather weak and even distribution throughout the disc. In contrast, staining for decorin was faint in the intermediate zone and the central part of the posterior band, moderate in the anterior and posterior attachments and most intense in the junction between the anterior band and attachment. The upper surface of the disc stained more intensely than the lower. Similarly, there was intense staining for large chondroitin-sulphate proteoglycan in the peripheral band, but both the anterior and the temporal parts of the posterior attachments were faintly stained. These results demonstrate marked regional differences in the expression of biglycan, decorin and large chondroitin-sulphate proteoglycan in the temporomandibular joint discs of adult rats. These variations probably reflect the different biomechanical environments caused by the complicated articulatory functions of the temporomandibular joint.

Animals↗

Epithelial rests colocalize with cementoblasts forming acellular cementum but not with cementoblasts forming cellular cementum.

Epithelial rests of Malassez and cementoblasts were examined in the rat molars during the early stages of root formation using an antilaminin antibody and/or peanut agglutinin (PNA), and an antiosteocalcin (OC) antibody, respectively. The roots of the first molars were used for study. The antilaminin antibody stained the basement membrane surrounding the epithelial root sheath and epithelial rests. The basement membrane of the epithelial root sheath was continuous, but that of the epithelial rests was discontinuous. The cells of epithelial rests and epithelial root sheath were positive for PNA. The structural characteristics of the epithelial rests were seen in the sections stained doubly with PNA and the antilaminin antibody. The cells of epithelial rests were fibroblast-like and formed a fine mesh in 2-week-old rats. In 3-week-old rats, the epithelial rests were also present at the coronal half of root surface, showing typical cell cords, but were not present at the apical part of the root surface where the cellular cementum covered the root dentin. At the root apex of 3-week-old rats, the cells of epithelial rests forming fine meshes were seen near the epithelial root sheath. The anti-OC antibody stained cementoblasts lining acellular and cellular cementum. The sections doubly stained with the anti-OC and the antilaminin antibodies or PNA further revealed the close relation between epithelial rests and cementoblasts. The OC-positive cells lining acellular cementum or dentin were localized very close to the epithelial rests. In contrast, the OC-positive cells lining cellular cementum did not show close association with the epithelial cells, except the cells located most apically where the basement membrane of the epithelial root sheath is disrupted and the initial cellular cementum begins to be formed. The present results suggest that the epithelial rests and/or the discontinuous basement membrane of them may have a role for the acellular cementum formation at least in the early stage of root formation.

Animals↗

Hypercoagulable state and disseminated intravascular coagulation following an effective chemotherapy in tumor-bearing rats.

OBJECTIVE: To detect the changes in blood coagulation system and clarify the related mechanisms of chemotherapy-induced disseminated intravascular coagulation. METHODS: Changes in blood coagulation system and immunohistochemistry for fibrinogen were investigated in six groups of rats designed for different purposes. RESULTS: Decreased platelet count, prolonged prothrombin time and active partial thromboplastin time, elevated fibrinogen level, and decreased antithrombin were observed in the rats receiving a newly developed chemotherapy (NDC group), in which mitomycin C was administered intravenously together with angiotensin. Accumulation of fibrinogen and microthrombi in the blood vessels of multiple organs were also found in the NDC group by immunohistochemistry and histopathological examination. CONCLUSIONS: Rapid reduction of tumor mass induced by an effective chemotherapy could cause hypercoagulable state and disseminated intravascular coagulation.

Angiotensins↗

Characterization of the fate of midline epithelial cells during the fusion of mandibular prominences in vivo.

The fusion of the mandibular prominences along the midline is achieved with the absence of medial epithelial cells at the fusion site. Failure of fusion of the mandibular prominences results in median cleft of the lower lip and mandible. Cellular and molecular events controlling mandibular fusion were examined during the fusion process in mouse embryogenesis. Cell lineage analyses at the fusion site revealed that epithelial cells migrated to the surface and oral epithelia. DiI-labeled epithelial cells were not observed within the mandibular mesenchyme at any state of fusion. Examination of the midline region did not reveal cells with ultrastructural changes characteristic of apoptotic cell death. An increase in lysosomal enzymes in the midline epithelial cells, which would be correlated with programmed cell death, was not observed. Mice lacking TGF-beta 3 did not have cleft mandible, but had clefting of the secondary palate as a feature of null mutation phenotype. We interpret our comparisons between wild type and homozygous TGF-beta 3 (-/-) mice to suggest that different developmental processes control palatal vs. mandibular fusion. We hypothesize that medical epithelial cells at the fusion site of mandibular prominences migrate to the surface epithelium during the fusion process and neither transdifferentiate into mesenchyme nor express apoptosis.

Animals↗

BMPs induce endochondral ossification in rats when implanted ectopically within a carrier made of fibrous glass membrane.

BACKGROUND: Bone morphogenetic proteins (BMPs) replicate the process of embryonic bone formation when implanted in ectopic sites. Our previous studies have indicated that BMPs can induce intramembranous ossification, i.e., direct bone formation without preexisting cartilage when implanted in rats subcutaneously by using the fibrous collagen membrane (FCM) as a carrier for implanting BMPs (Sasano et al. 1993. Anat. Rec., 236:373-380). The present study was designed to investigate how the physicochemical property of the carrier material influences the process of bone formation induced by BMPs, using a carrier made of fibrous glass membrane (FGM). METHODS: BMPs, partially purified from bovine metatarsal bones, were added to an FGM carrier and implanted subcutaneously in rats. The implants were analyzed at weekly intervals, and the osteogenic process induced by BMPs was examined by histology and immunohistochemistry for cartilage and bone formation. RESULTS: Neither cartilage nor bone were observed after week 1. Cartilage formation occurred within the carrier after week 2, although no bone formation was seen. The cartilage matrix showed immunoreactivity for types II, X, and I collagen. Bone was induced on the previously formed cartilage after week 3. The bone matrix stained with anti-osteocalcin antibody and with anti-type I collagen antibody. The cartilage was replaced by bone and bone marrow after week 10. CONCLUSIONS: BMPs cause endochondral ossification when administered with an FGM carrier. The physicochemical property of the carrier may be involved in the BMP-induced phenotype expression of bone and cartilage.

Animals↗

Localization of types I, II and III collagen and glycosaminoglycans in the mandibular condyle of growing monkeys: an immunohistochemical study.

In order to analyse the regional and age-related variations of primate condyles, immunohistochemical techniques were used to examine the localization of types I, II and III collagen and a variety of glycosaminoglycans in distinct anteroposterior regions of the mandibular condyle of two growing female rhesus monkeys (Macaca mulatta). In the juvenile monkey staining for types I and III collagen was weak in the fibrous tissue layer, intense in the pre-cartilaginous tissue layer and faint in the cartilaginous tissue layer; staining was significantly more intense in the posterosuperior and posterior regions than in the anterior region. Similarly, staining for cartilage-characteristic extracellular matrices, including type II collagen and keratan sulfate, was intense in the cartilaginous tissue layer of the posterior condyle. In contrast, in the late-adolescent monkey staining for the extracellular matrices was more intense in the anterior half of the condyle (i.e. from the anterior to the posterosuperior region) than in the posterior region, and most intense in the posterosuperior region. The results demonstrate that marked regional differences exist in the phenotypic expression of the extracellular matrices in the mandibular condyles of growing monkeys and that these differences vary between different developmental stages. The variations probably reflect the predominance of competing growth and articulatory functions in the mandibular condyles.

Aging↗

Confocal microscopy of cementocytes and their lacunae and canaliculi in rat molars.

The present study was designed to analyze the morphological characteristics of cementocytes and osteocytes. The maxillae of 10-week-old Wistar rats were used for observations. Non-decalcified ground sections stained vitally with fluorescence dyes and decalcified frozen sections stained with FITC-phalloidin were examined by confocal microscopy. Calcein and alizarin red stained the calcification front of bone, cementum, and dentin intensely. In addition, lacunae and canaliculi of cementocytes and osteocytes as well as dentinal canals were stained with the fluorescent dyes. The staining of lacunae and canaliculi was less intense than that of the calcification front of bone, cementum and dentin. The canaliculi of cementocytes and osteocytes were connected with the canaliculi extending from the calcification front of cementum and bone, respectively. The canalicular density was less in the cellular cementum than in the bone. Areas devoid of canaliculi were numerous in the cellular cementum, whereas areas devoid of canaliculi were scarce in the alveolar bone. Further, the lacunae of cementocytes showed various shapes, from oval to tubular, while the lacunae of osteocytes were invariably oval. The cell body and the cytoplasmic processes of cementocytes were positive for FITC-phalloidin within the extracellular matrix of cellular cementum, which was negative. The distribution of actin filaments in the osteocytes and the cementocytes was predominantly cortical and appeared to be closely associated with the cell membrane of the cell bodies and the cytoplasmic processes. Intense staining was seen at the proximal part of the cytoplasmic processes in both osteocytes and cementocytes, showing a punctuated structure of the cells that was more frequent in osteocytes than in cementocytes. The stress fiber known to be present in most of the cultured cells was not evident in the these cells in situ. The cells incorporated in the cementodentinal junction were strongly stained with FITC-phalloidin. The distribution pattern of the cytoplasmic processes stained with FITC-phalloidin was similar to that of the canaliculli stained vitally. The cytoplasmic processes of osteocytes and cementocytes were connected with those of cells lining the surface of bone and cementum. The present result-that lacunae and canaliculi of cementocytes were stained vitally with the fluorescence dyes-suggests that cementocytes may have a role in secondary calcification of cellular cementum. Further, the lower density of cytoplasmic processes in cementocytes than in osteocytes suggests a lack of complexity in the intercellular network within the cellular cementum.

Actins↗

Localization of types I, II and X collagen and osteocalcin in intramembranous, endochondral and chondroid bone of rats.

Chondroid bone is a unique calcified tissue intermediate between bone and cartilage. To clarify its characteristics, we examined the distributions of the ECMs associated with chondrogenic differentiation and matrix calcification in the chondroid bone of the rat glenoid fossa, and compared them to those in two typical bone tissues, alveolar bone of the maxilla (intramembranous bone) and the growth plate of long bone (endochrondral bone), using immunofluorescence techniques. Morphologically, the glenoid fossa consisted of the fibrous, progenitor and cartilaginous cell layers and the cartilaginous cell layer was further divided into the superficial non-hypertrophic layers (secondary cartilage) and the deep hypertrophic cell layers (chondroid bone). The co-distribution of type I and type II collagens was observed in secondary cartilage and chondroid bone, whereas type X collagen was restricted to the pericellular matrix of hypertrophied cells (chondroid bone). Osteocalcin, which was absent from the calcified cartilage of endochondral bone formation, was also present in the ECM of the chondroid bone, but not in cells. These results demonstrate that chondroid bone of rats, which is adjacent to secondary-type cartilage in the glenoid fossa, has phenotypic expressions associated with both hypertrophied chondrocytes and osteocytes.

Animals↗

Development of interglobular dentine in rat molars and its relation to maturation of enamel.

The development of interglobular dentine in the first upper and lower molars of Wistar rats aged 3, 7, 14, 21, 42 days was examined histochemically using a lectin, succinyl wheat germ agglutinin (sWGA), which is specific for N-acetyl-D-glucosamine. sWGA stained the interglobular dentine, predentin and Golgi area of odontoblasts. Interglobular dentine was not formed in the first molars of 3-day rats, but appeared in those of 7-day rats near the enamel-free area. In 14-day rats, interglobular dentine was present in most areas of the coronal dentine except the cervical area. At the interface between dentine and predentin, numerous sWGA-negative calcospherites were seen, suggesting that the interglobular dentine is formed actively there. In 21-day rats, the interlobular dentine was more numerous than in 14-day rats. Interglobular dentine was present in the cervical root dentine as well as in the coronal dentine, including the cervical area. The distribution of interglobular dentine in 42-day rats was similar to that in 21-day rats, but fluorescence of sWGA binding was less intense in the former. Because the development of interglobular dentine appeared to be time and position specific its relation to the stages of ameloblasts was analysed. Thin enamel matrix was formed at cusps in molars of 3-day rats and thickness of enamel matrix increased in 7-day rats. In these teeth, the ameloblasts were at the differentiating or secretory stage. The Golgi area and Tomes' processes of the secretory ameloblasts, the cells of intermediate layer and the enamel matrix were weakly positive with sWGA. The epithelial cells at the enamel-free area were also stained with sWGA. In 14-day rats, most of the ameloblasts in the first maxillary molars were at the maturative stage except in the cervical area, where the ameloblasts were at the transitional stage. sWGA stained the distal border and the Golgi area of the maturative ameloblasts as well as the cells of the papillary layer. The distal border of the maturative ameloblasts appeared either thick or thin, suggesting a ruffle-end and smooth-end of the cells. Ameloblasts were absent in the first molars of 21-day rats and the cervical part of the enamel was covered with the stratified epithelium like that of 42-day rats. The present study has demonstrated that interglobular dentine contains sWGA-binding glycoconjugates and the formation of the interglobular dentine is largely associated with the enamel maturation. These results suggest that matrix-to-cell interaction is important for the development of interglobular dentine.

Acetylglucosamine↗

Multinucleated giant cells recruited by implantation of octacalcium phosphate (OCP) in rat bone marrow share ultrastructural characteristics with osteoclasts.

The ultrastructural characteristics of multinucleated giant cells (MNGCs) on octacalcium phosphate (OCP) and hydroxyapatite (HA) were investigated in comparison with those of osteoclasts, when the synthetic OCP and HA were implanted in rat bone marrow. The morphological difference in the MNGCs were shown between OCP and HA implants in 2 weeks after implantation. The MNGC on the implanted OCP (i-OCP) developed the ruffled border-like structure and the clear zone-like structure. The i-OCP was frayed where it was in contact with the ruffled border-like structure. The MNGC on the implanted HA (i-HA) developed the clear zone-like structure, whereas no ruffled border was seen. The surface of i-HA associated with the MNGC was smooth and not frayed. Bone formed directly on the OCP or HA implants. The interface between the i-OCP and bone matrix interdigitated, whereas that between the i-HA and bone matrix was comparatively smooth. The present study suggested that the i-OCP could be resorbed by the MNGCs which share some ultrastructural characteristics with osteoclasts.

Animals↗

Expression of osteocalcin in cementoblasts forming acellular cementum.

To determine the phenotypic expression of cementoblasts responsible for acellular cementum, an immunohistochemical study was performed using a polyclonal antibody raised against the aminoterminal peptide of rat osteocalcin (OC). Maxillary first molars of Wistar male rats aged 2 and 3 wk were used for observations. Serial sections of decalcified paraffin embedded specimens were stained either with hematoxylin and eosin or with the anti-OC antibody. In 2-wk-old rats, apical roots were lined with the epithelial root sheath. A thin layer of acellular cementum was seen at most of the root surface, but was not seen near to root apex. In 3-wk-old rats, cellular cementum began to be formed at root apex, and acellular cementum became more thick than in 2-wk-old rats. Acellular and cellular cementum were lined with the fibroblast-like cells. Osteocalcin staining was detected in cells lining root surface in both 2- and 3-wk-old rats. Almost all cells lining cellular cementum were positive for OC. In contrast OC positive cells lining acellular cementum and root surface devoid of cementum appeared at a specific site of the root. The cells at the interradicular area of root surface were positive but the cells at the outer area (the opposite side of the interradicular area) were negative for OC. Osteoblasts and odontoblasts were positive with the antibody. The present results suggest that the OC expression of cementoblasts forming acellular cementum is similar to that of cells forming cellular cementum as well as osteoblasts and odontoblasts, and has a role for calcification of acellular cementum.

Animals↗

Age-related changes in the localization of glycosaminoglycans in condylar cartilage of the mandible in rats.

There is little information available regarding the morphological and biomolecular characteristics of mandibular condylar cartilage. The purpose of this study was to determine the age-related changes in the morphology and immunolocalization of glycosaminoglycans (GAGs) in mandibular condyles. The mandibular condylar cartilages from 4-, 8-, 16-, 32-, and 64-week-old Wistar male rats were examined to verify the localization of chondroitin-4-sulfate (Ch-4S), chondroitin-6-sulfate (Ch-6S) and keratan sulfate (KS) using an indirect immunofluorescent technique with three monoclonal antibodies for glycosaminoglycans, 2-B-6, 3-B-3 and 5-D-4, respectively. Morphologically, the condylar cartilage was a growth cartilage during growing periods, began to differentiate into articular cartilage from the central area of 16-week-old condyles, and became mature articular cartilage at 32 weeks of age. A regional difference was found in the morphological features and distribution of GAGs between the anterior, central, postero-superior and posterior areas of the condyles at each age. The immunohistochemical localizations of these three glycosaminoglycans showed age-related, morphology-dependent changes, from growth cartilage to articular cartilage-like cartilage. Immunoreactions for all of the antibodies decreased progressively with age in the interterritorial matrix, while the pericellular and territorial matrix in the condylar cartilage of the mandible maintained relatively higher immunoreactivity. In conclusion, age-related and regional differences in the localization of glycosaminoglycans Ch-4S, Ch-6S, and KS were found in the mandibular condyles in rats, and these changes are believed to be related to functional and developmental requirements.

Aging↗