Biomedical subjects

Y Sasano

Publications and source records attributed to Y Sasano.

At least 19 recordsLinked to original sources

PubMed2006

Characterization of dentin formed in transplanted rat molars by electron probe microanalysis.

The present study was designed to characterize dentin formed in transplanted rat molars by investigating calcium (Ca), phosphorus (P), and magnesium (Mg) concentrations using electron probe microanalysis (EPMA) as well as examining the rate of dentin matrix formation by vital staining. The unerupted immature lower right second molar in 2-week-old rats was transplanted into the upper right first molar socket. Rats were injected with oxytetracycline, calcein, and alizarin intraperitoneally at 1 day before and 1 and 2 weeks after transplantation, respectively, for vital staining. The maxillae and mandibles were fixed 3 weeks after transplantation, resected, and embedded in resin. Undemineralized sections were cut and examined by fluorescent microscopy and EPMA. The thickness of dentin formed in the first week after transplantation was significantly less than that of dentin formed in any other 1-week period in the transplanted tooth and was about one-fifth the thickness of dentin formed in control teeth. Formation of dentin recovered in the third week after transplantation. In the first week after transplantation, EPMA demonstrated a sharp increase in Mg concentration with a slight decrease in Ca concentration. Thereafter, no significant difference was identified among Ca, P, or Mg concentrations or the Ca/P ratio between transplanted and control teeth. These results suggest that disruption of the circulation and innervation by transplantation cause a temporary change in the matrix formation rate and elemental distribution of dentin, which is subsequently restored within 2 weeks after transplantation.

Animals↗

PubMed2005

Osteoclastogenic activity during mandibular distraction osteogenesis.

Mandibular distraction osteogenesis is a well-developed clinical modality for the treatment of craniofacial deformities and dental arch discrepancies, in combination with orthodontic treatment. However, in our previous study, orthodontic tooth movement into the distraction gap caused severe root resorption. The present study aimed to clarify the osteoclastogenic activity of cells in the distraction gap. We hypothesized that the gene expression of osteoclastogenic- and osteoclast-supporting molecules in osteoblasts and stromal cells would increase at distraction sites during the consolidation period. An animal model experiment involving rabbits was designed for mandibular distraction osteogenesis and subjected to in situ hybridization analysis. The number of osteoclasts was larger in the distraction gap during the early consolidation period than in normal controls, due to an increase of gene expression for osteoclastogenic cytokines in osteoblasts. It was concluded that osteoclastogenic and osteoclastic activities are stimulated at distraction sites during the early consolidation period.

Animals↗

PubMed2004

New scaffold for recombinant human bone morphogenetic protein-2.

A bioactive and resorbable scaffold is necessary to exhibit the osteoinductive potency of recombinant human bone morphogenetic protein-2 (rhBMP-2). In a previous study, we found that synthetic octacalcium phosphate (OCP) enhances bone regeneration and is replaced by newly formed bone after it is resorbed. We hypothesized that OCP may be useful as an effective scaffold for rhBMP-2 to enhance bone regeneration. To test this hypothesis, the present study was designed to investigate whether an OCP/BMP composite implant could more effectively enhance bone regeneration. A critical-sized defect was made in a rat calvarium and 1. 15 mg of OCP combined with 10 microg of rhBMP-2 (OCP/BMP 10 microg), 2. 15 mg of OCP combined with 1 microg of rhBMP-2 (OCP/BMP 1 microg), or 3. OCP (OCP alone) was implanted into the defect and fixed at 4 or 8 weeks after implantation. The percentage of newly formed bone (n-Bone%) in the defect was determined by a histomorphometrical analysis. A statistical analysis showed that n-Bone% with OCP/BMP was significantly higher than that with OCP at both time points, whereas the difference in n-Bone% between OCP/BMP 10 microg and OCP/BMP 1 microg was not significant. The present results suggest that OCP can be used as an effective scaffold for rhBMP-2 and this OCP delivery system may be able to reduce the standard effective dose of rhBMP-2, which would be beneficial because low doses (<100 microg/g OCP) of rhBMP-2 enhance bone regeneration.

Absorbable Implants↗

PubMed2003

Expression of MMP-8 and MMP-13 genes in the periodontal ligament during tooth movement in rats.

Periodontal ligament tissue is remodeled on both the tension and compression sides of moving teeth during orthodontic tooth movement. The present study was designed to clarify the hypothesis that the expression of MMP-8 and MMP-13 mRNA is promoted during the remodeling of periodontal ligament tissue in orthodontic tooth movement. We used the in situ hybridization method and semi-quantitative reverse-transcription/polymerase chain-reaction analysis to elucidate the gene expression of MMP-8 and MMP-13 mRNA. Expression of MMP-8 and MMP-13 mRNA transiently increased on both the compression and tension sides during active tooth movement in vivo. The gene expression of MMP-8 and MMP-13 was induced by tension, while compression indirectly promoted the gene expression of MMP-8 and MMP-13 through soluble factors in vitro. Thus, we concluded that the expression of MMP-8 and MMP-13 is differentially regulated by tension and compression, and plays an important role in the remodeling of the periodontal ligament.

Animals↗

PubMed2002

Implanted octacalcium phosphate is more resorbable than beta-tricalcium phosphate and hydroxyapatite.

Our previous studies have suggested that synthetic octacalcium phosphate (OCP) could be resorbed and replaced by newly formed bone if implanted in rat skull defects. We hypothesized that the implanted OCP is more resorbable than other commonly used bone graft substitutes of calcium phosphate compounds, such as hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP). To test the hypothesis, the present study was designed to compare histomorphometrically resorption of the implanted OCP, HA, and beta-TCP, which were kept in the experimental cranial defect of rats for a long term. A full thickness of standardized trephine defect was made in the rat parietal bone, and the same volume of granules of OCP, HA, and beta-TCP were implanted into the defect. Five specimens of each group were fixed 6 months after implantation. The percentage of remaining implants (r-Imp%) and newly formed bone (n-Bone%) in the defect was analyzed histomorphometrically. The statistical analysis showed that the r-Imp% of OCP was significantly lower than that of HA and beta-TCP. In contrast, the n-Bone% of OCP was significantly higher than that of HA and beta-TCP. The present study has shown that the implanted OCP in the rat cranial defect is more resorbable than the implanted beta-TCP and HA, whereas the implanted OCP enhances bone formation more than the implanted beta-TCP and HA.

Animals↗

PubMed2002

Expression of MMP-8 and MMP-13 mRNAs in rat periodontium during tooth eruption.

The present study was designed to investigate mRNA expression of matrix metalloproteinase-8 (MMP-8) and MMP-13 in forming periodontium during tooth eruption in the rat. RT-PCR for the decalcified paraffin sections indicated expression of MMP-8 and MMP-13 in the periodontal tissues. In situ hydridization demonstrated expression of MMP-8 in osteoblasts, osteocytes, periodontal ligament cells, cementoblasts, and cementocytes along with collagen types I and III. In contrast, transcripts of MMP-13 were confined to a small population of osteoblasts and osteocytes in alveolar bone. The results suggested that MMP-8 may be involved in remodeling the periodontium during tooth eruption, and its expression may be coordinated with that of collagen types I and III, whereas the participation of MMP-13 may be rather limited.

Alveolar Process↗

PubMed2001

Osteoblastic differentiation of periosteum-derived cells is promoted by the physical contact with the bone matrix in vivo.

The periosteum contains osteoprogenitors that differentiate to osteoblasts in bone growth or repair. Our previous studies suggested the hypothesis that the physical contact of the periosteum with the bone matrix is requisite for the differentiation of osteoblasts. To test the hypothesis, the present study was designed to investigate how the contact between the periosteum and the bone matrix influences the osteoblastic differentiation of periosteal cells with establishing a new experimental model in vivo. Differentiation of osteoblasts was assessed by gene expression of type I collagen, osteocalcin and bone sialoprotein using in situ hybridization. A barrier was designed to prevent periosteal cells from contacting the bone matrix using the membrane filter. The membrane filter was inserted surgically between the surface of rat parietal bone and the periosteum after being punched out with pin holes. Periosteal cells were allowed to contact with the bone surface only through the pin holes. The pin hole was filled with cells derived from the periosteum 1 week after inserting the filter. Differentiation of osteoblasts in week 2 and noticeable bone formation in week 3 were identified on the bone surface only under the pin hole but not under the filter. The present study demonstrated that the physical contact with the bone matrix promotes osteoblastic differentiation of periosteum-derived cells in vivo.

Animals↗

PubMed2001

Implantation of octacalcium phosphate combined with transforming growth factor-beta1 enhances bone repair as well as resorption of the implant in rat skull defects.

In our previous study, we reported that synthetic octacalcium phosphate (OCP) enhances bone repair if implanted in rat skull defects. We hypothesized that OCP can be used as an effective carrier for transforming growth factor-beta1 (TGF-beta1) to promote bone repair. We designed the present study to investigate histomorphometrically whether combination with recombinant human TGF-beta1 could promote bone repair caused by OCP per se (Control/OCP). A full-thickness standardized trephine defect was made in the rat parietal bone and OCP combined with recombinant human TGF-beta1 (TGF-beta1/OCP) or Control/OCP was implanted into the defect. Four rats from each group were fixed at 2, 4, and 8 weeks after implantation. Histomorphometrical analysis of the percentage of newly formed bone (n-Bone %) and remaining implants (r-Imp %) in the defect was performed. The statistical analysis showed the n-Bone % of TGF-beta1/OCP was significantly higher than that of the Control/OCP in week 4, whereas the r-Imp % of TGF-beta1/OCP was significantly lower than that of the Control/OCP. The present study demonstrated that OCP can be used as an effective carrier for TGF-beta1 and their combination enhances bone repair as well as resorption of the carrier OCP in the early stage of bone formation.

Animals↗

PubMed2001

Characterization of interglobular dentin and Tomes' granular layer in dog dentin using electron probe microanalysis in comparison with predentin.

The interglobular dentin (IG) and the Tomes' granular layer (TGL) as well as predentin are hypomineralized regions in dentin. Some previous studies proposed that the IG and the TGL are identical with difference only in size, whereas other suggested that they are distinct structures. In order to characterize their matrix components, the present study was designed to analyze the elements of calcium (Ca), phosphorus (P), and sulfur (S) in the IG and the TGL in comparison with predentin using the Electron Probe Microanalysis (EPMA). The TGL was highest in the concentration of both Ca and P among the hypomineralized regions followed by the IG and predentin, whereas predentin was the highest in the concentration of S followed by the IG and the TGL. Alcian blue staining suggested that the S elements identified with the EPMA are incorporated into the sulfated glycosaminoglycan chains of proteoglycans. The present study first demonstrated distinct characteristics of matrix components in the IG and the TGL, i.e., the IG is poorer in mineralization but much richer in a proteoglycan content than the TGL. The IG may originate from predentin because of their analogy, whereas the TGL may follow a different ontogeny.

Alcian Blue↗

PubMed2001

Temporal and spatial gene expression of major bone extracellular matrix molecules during embryonic mandibular osteogenesis in rats.

It is not known how gene expression of bone extracellular matrix molecules is controlled temporally and spatially, or how it is related with morphological differentiation of osteoblasts during embryonic osteogenesis in vivo. The present study was designed to examine gene expressions of type I collagen, osteonectin, bone sialoprotein, osteopontin, and osteocalcin during mandibular osteogenesis using in situ hybridization. Wistar rat embryos 13-20 days post coitum were used. The condensation of mesenchymal cells was formed in 14-day rat embryonic mandibles and expressed genes of pro-alpha 1 (I) collagen, osteonectin, bone sialoprotein and osteopontin. Cuboidal osteoblasts surrounding the uncalcified bone matrix were seen as early as in 15-day embryonic mandibles, while flat osteoblasts lining the surface of the calcified bone were seen from 16-day embryonic mandibles. Cuboidal osteoblasts expressed pro-alpha 1(I) collagen, osteonectin and bone sialoprotein intensely but osteopontin very weakly. In contrast, flat osteoblasts expressed osteopontin very strongly. Osteocytes expressed the extracellular matrix molecules actively, in particular, osteopontin. The present study demonstrated the distinct gene expression pattern of type I collagen, osteonectin, bone sialoprotein, osteopontin and osteocalcin during embryonic mandibular osteogenesis in vivo.

Animals↗

PubMed2001

Distinctive expression of extracellular matrix molecules at mRNA and protein levels during formation of cellular and acellular cementum in the rat.

Little is known about differential expression of extracellular matrices secreted by cementoblasts between cellular and acellular cementum. We hypothesize that cementoblasts lining acellular cementum express extracellular matrix genes differently from those lining cellular cementum, thereby forming two distinct types of extracellular matrices. To test this hypothesis, we investigated spatial and temporal gene expression of selected extracellular matrix molecules, that is type I collagen, bone sialoprotein, osteocalcin and osteopontin, during formation of both cellular and acellular cementum using in situ hybridization. In addition, their extracellularly deposited and accumulated proteins were examined immunohistochemically. The mRNA transcripts of pro-alpha1 (I) collagen were primarily localized in cementoblasts of cellular cementum and cementocytes, while those of bone sialoprotein were predominantly seen in cementoblasts lining acellular cementum. In contrast, osteocalcin was expressed by both types of cementoblasts and cementocytes and so was osteopontin but only transiently. Our immunohistochemical examination revealed that translated proteins were localized extracellularly where the genes had been expressed intracellularly. The present study demonstrated the distinctive expression of genes and proteins of the extracellular matrix molecules between cellular and acellular cementum.

Animals↗

PubMed2001

Gene and protein expressions of type I collagen are regulated tissue-specifically in rat hyaline cartilages in vivo.

The present study was designed to investigate how rat hyaline cartilages at various sites in vivo express the gene and protein of type I collagen using in situ hybridization and immunohistochemistry. The gene of pro alpha 1(I) collagen was expressed by chondrocytes in articular cartilage, and the protein of type I collagen was identified in the cartilage matrix. In contrast, growth plate cartilage expressed the gene of pro alpha 1(I) collagen, but no protein of type I collagen. Neither gene nor protein of type I collagen was expressed in cartilages of trachea and nasal septum. The present study suggested that expression of type I collagen in hyaline cartilages may be regulated tissue-specifically at gene and/or protein levels.

Animals↗

PubMed2001

Implantation of octacalcium phosphate nucleates isolated bone formation in rat skull defects.

OBJECTIVE: Our previous radiographic examinations have indicated that the synthetic octacalcium phosphate (OCP) may provide the core for nucleating multiple osteogenic sites in the experimentally created cranial defect. DESIGN: The present study was designed to confirm the possibility that the implanted OCP causes the osteoinduction as well as the osteoconduction in the rat cranial defect. MATERIALS AND METHODS: Standardized defects were created in male Wistar rat calvaria, and the OCP granules were implanted into the defect. The sham operated rats were processed in the same way except that nothing was implanted. The rats were fixed at 4 weeks after implantation of OCP or the sham operation. We examined bone formed on the implanted OCP, analyzing serial sections histologically combined with immunohistochemistry for the bone specific protein, osteocalcin. RESULTS: In the defects treated with OCP, the radiopacity was scattered throughout the defect besides being observed along the defect margin of the parietal bone. Examination of the serial sections showed that some of new bones on the implanted OCP were formed away from the defect margin of the parietal bone with regard to both histological identification and specific molecular marker. CONCLUSIONS: The present study suggested that the implanted OCP can serve as a core for initiating bone formation and cause the osteoinduction as well as the osteoconduction in the defect.

Animals↗

PubMed2000

Compressive force promotes chondrogenic differentiation and hypertrophy in midpalatal suture cartilage in growing rats.

Midpalatal suture cartilage (MSC) is secondary cartilage located between the bilateral maxillary bones and has been utilized in the analysis of the biomechanical characteristics of secondary cartilage. The present study was designed to investigate the effects of compressive force on the differentiation of cartilage in midpalatal suture cartilage in rats. Forces of various magnitudes were applied to the midpalatal suture cartilage in 4-week-old male Wistar rats for 1, 2, 4, 7, or 14 days, mediated through the bilateral 1st molars using orthodontic wires. The differentiation pathways in the MSC cells were examined by immunohistochemistry for the differentiation markers type I, type II and type X collagen, and glycosaminoglycans (GAGs), chondroitin-4-sulfate, chondroitin-6-sulfate and keratan sulfate. Histologically and immunohistochemically, the midpalatal suture cartilage in control rats had the characteristic appearance of secondary cartilage. In the experimental groups, the center of the midpalatal suture cartilage that contained osteo-chondro progenitor cells seemed to become mature cartilage and its immuno-reaction to type II and X collagen and GAGs increased as the experiment progressed. This differentiation was dependent upon the magnitude and duration of the force applied to the midpalatal suture cartilage; i.e., cartilaginous differentiation progressed more rapidly as the applied force increased. The present results suggest that the differentiation of osteo-chondro progenitor cells into mature and hypertrophic chondrocytes in the precartilaginous cell layer is promoted by compressive force.

Animals↗

PubMed2000

Confocal microscopy of Tomes' granular layer in dog premolar teeth.

Tomes' granular layer is the hypomineralized area of radicular dentin, but knowledge concerning it is limited. The present study was designed to investigate the structural characteristics of Tomes' granular layer in the dog's teeth by confocal microscopy. Permanent premolars of four beagles, two at 7 months and the other two at 14 months of age, were used for observation. During premolar root formation, the 7-month-old dogs were injected with calcein and alizarin red S for vital staining of dentin, and ground sections of the teeth were prepared. Both ground and decalcified-paraffin sections were made from the teeth of the 14-month-old dogs and stained with basic fuchsin or with hematoxylin and eosin. All sections were examined by fluorescence and confocal microscopy. In the ground sections, granules of Tomes' layer and dentinal tubules were stained with basic fuchsin and with calcein. The granules of Tomes' layer stained with calcein were seen only near the labeling lines by calcein. The granules of Tomes' layer appeared as bright spots in cross sections, and as lines in longitudinal sections. When the sections were cut tangentially through the surface of dentin, the granules of Tomes' layer showed a reticular structure. Most of the dentinal tubules were seen to pass between the granules and terminated in the dentin-cementum junction. Looped tubules were not found in this area. In the paraffin sections stained with hematoxylin and eosin, extracellular matrix of dentin showed fluorescence of various intensities and dentinal tubules appeared dark. At the surface of the radicular dentin, the granules of Tomes' layer appeared as fluorescent fibers running parallel to the surface of dentin in the longitudinal sections. The fibers appeared as bright spots in the cross sections and as a mesh in the tangential sections. In the periodontal ligament, collagen fibers showed intense fluorescence, whereas most cells were negative. From these results we conclude that Tomes' granular layer of dog's teeth may be the collagen fiber bundles that remained uncalcified or hypocalcified within the radicular dentin.

Aging↗

PubMed2000

Expression of major bone extracellular matrix proteins during embryonic osteogenesis in rat mandibles.

It is not known how bone proteins appear in the matrix before and after calcification during embryonic osteogenesis. The present study was designed to investigate expressions of the five major bone extracellular matrix proteins--i.e. type I collagen, osteonectin, osteopontin, bone sialoprotein and osteocalcin--during osteogenesis in rat embryonic mandibles immunohistochemically, and their involvement in calcification demonstrated by von Kossa staining. Wistar rat embryos 14 to 18 days post coitum were used. Osteogenesis was not seen in 14-day rat embryonic mandibles. Type I collagen was localized in the uncalcifed bone matrix in 15-day mandibles, where no other bone proteins showed immunoreactivity. Osteonectin, osteopontin, bone sialoprotein and osteocalcin appeared almost simultaneously in the calcified bone matrix of 16-day mandibles and accumulated continuously in 18-day mandibles. The present study suggested that type I collagen constitutes the basic framework of the bone matrix upon which the noncollagenous proteins are oriented to lead to calcification, whereas the noncollagenous proteins are deposited simultaneously by osteoblasts and are involved in calcification cooperatively.

Animals↗

PubMed2000

Immunohistochemical localization of type I collagen, fibronectin and tenascin C during embryonic osteogenesis in the dentary of mandibles and tibias in rats.

Type I collagen, fibronectin and tenascin C play an important role in regulating early osteoblast differentiation, but the temporal and spatial relationship of their localization during embryonic osteogenesis in vivo is notknown. The present study was designed to localize these three molecules in the dentary of mandibles and tibias in rat embryos using immunohistochemistry. Serial paraffin sections were cut and adjacent sections were processed for von Kossa staining or immunohistochemistry for type I collagen, fibronectin and tenascin C. In the dentary, tenascin C was localized within and around the mesenchymal cell condensation in embryos at 14 days in utero. The bone matrix at 15 days showed immunoreactivity for both type I collagen and fibronectin. The immunoreactivity of type I collagen was persistent, whereas that of fibronectin decreased with age of embryos. In tibias, tenascin C was localized in the perichondral mesenchymal tissue at 17 days. Immunoreactivity for type I collagen was persistent in the bone matrix, whereas the tibial bone showed little immunoreactivity for fibronectin at any embryonic age examined. The present study demonstrated characteristic localization of type I collagen, fibronectin and tenascin C during embryonic osteogenesis in the dentary of mandibles and tibias.

Animals↗

PubMed2000

Distribution of interglobular dentine in human tooth roots.

The present study was designed to examine the distribution of interglobular dentine in human tooth roots. The material comprised 17 teeth, of which 3 were premolars extracted for orthodontic reasons from children 10-12 years of age and the other teeth (4 incisors, 3 canines and 7 molars) were extracted for periodontitis from individuals aged 32-63 years. All teeth were free of caries and cervical dentine defects. Ground sections of the teeth cut longitudinally were stained with basic fuchsin and observed by fluorescence and confocal microscopy as well as transmitted light microscopy. Basic fuchsin stained the dentinal tubules, interglobular dentine and the granular layer of Tomes. These structures appeared intense blue to faint violet with transmitted light microscopy, whereas their staining displayed intense fluorescence with fluorescence microscopy. Therefore, the interglobular dentine could be detected more sensitively with fluorescence and confocal microscopy than with transmitted light microscopy. Typical interglobular dentine was present in coronal dentine in most of the teeth. In the radicular dentin, position and size of the interglobular dentine was different among the teeth examined. Most of the teeth had the interglobular dentine in the cervical part of the roots (type A). Two premolars displayed the interglobular dentine in the coronal half of the root (type B). The types A and B contained large interglobular areas. A small amount of interglobular dentine was restricted to the apical half of the roots of two canines and one molar (type C). In contrast to types A and B which were seen at both labial or buccal and lingual sides of roots, the interglobular dentine of type C was seen only at one side, labial or lingual. Some of the tooth roots did not show any interglobular dentine (type D). Most of the incisors, canines and premolar were types A, B, and C, respectively, and the molars were mixed types A, C, and D. These results suggest that the factors affecting dentinogenesis during root formation are unique for each tooth.

Adult↗