| Co-Investigator(Kenkyū-buntansha) |
TAJIMA Kenji Hokkaido Univ., Graduate School of Engineering, Associate Professor, 大学院・工学研究科, 助教授 (00271643)
SATOH Yasuharu Hokkaido Univ., Graduate School of Engineering, Assistant Prof., 大学院・工学研究科, 助手 (30360928)
NISHIMURA Kazuaki Osaka Dental Univ., Dep.of Periodontics, Lecturer, 歯学部, 講師 (40098017)
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| Research Abstract |
1.Purification and characterization of human periodontal ligment (hPDL) cell chemotactic facter from bovine teeth precementum : We reported that the composite cementum particles-embedded gelatine membrane induced root surface mesenchymal cells to differentiated into cementoblasts and a promotive adjunct for true guided tissue regeneration (formoing new cementum, new ligment, and new bone) following periodontal flap surgery. The purpose of this sudy was to investigate that characterization of chemotactic factors released from cementum. They were purified from PBS(-)extracts of bovine teeth precementum using gel filtration-HPLC,DEAE and CM-ionexchange-HPLC, Affinity HPLC, and SDS-PAGE. After the purification by SDS-PAGE, extracts bands were analyzed TOF-MAS. We obtained two new subutances (named CDCF-I,II) which were not related attachment and growth protein such as BSP,CAP,PDGF-BB,IGF-I,TGF-β and fibronectin.
2.Effects of irradiation on cementum matrix cytokines function during periodont
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al regeneration : The influence of γ-ray irradiation of cementum (γC) was analyzed with emphasis on its function during periodontal regeneration. With γC protein, sample cells (gingival fibroblasts, periodontal ligament cells, and alveolar bone cells) were co-cultured, and cytological parameters were analyzed. Additionally, kinetics of some gene expression was analysed using reverse transcript RT-PCR, which induced osteoprotegerin (OPG)/osteoclastogenesis inhibitoryt factor (OCIF) mRNA. The expression of OPG/OCIF mRNA was lower in co-cultured cells with γC protein than those with in control C protein (non-irradiated cementum). Together the results imply that some cytokine in intact cementum prevents the attachment (differentiation) of bone cells onto the root surface, which may explain why the introduction of CGM (cementum-impregnated gelatine membrane) following gingival flap surgery induced new cementum, new ligament and new bone formation, but CGM irradiated with γrays for clinical use causes ankylosis.
3.New arifficial periodontal ligament from salmon atelocollagen : The fish collagen has lower risk for transmission of infectious diseases to human beings than bovine collagen. We therefore consider that fish collagen could be an alternative to bovine collagen for use as biomaterials. The purpose of this study was to investigate the application of salmon atelocollagen (SAC) to a scaffold. SAC has a low denaturation temperature (Td:19℃ and needs to be cross-linked before being used as a scaffold. SAC was cross-linked by EDC. The material properties of the SAC scaffolds cross-linked by EDC were evaluated. It was found that EDC-SAC had a high degree of cross-linking and high stability(Td:55℃). Human periodontal ligament (hPDL) cells were cultured in the scaffolds for 2 weeks in vitro, and the activities(proliferation rate and alkaline phosphatase activity (ALP) ) of hPDL cells cultured in EDC-SAC were compared with those cultured in bovine atelocollagen (BAC) scaffolds cross-linked by EDC (EDC-BAC). The proliferation rate of hPDL cells cultured in EDC-SAC was equivalent to that in EDC-BAC, and the ALP activity in EDC-SAC was found to be significantly higher than that in EDC-BAC. There was no difference in the thermal stability, porous structure, and cell proliferation rate between EDC-SAC and EDC-BAC. In addition, the collagen helix of EDC-SAC was found to be partially denatured, and this structure resulted in the enhancement of ALP activity of hPDL cells compared with that using EDC-BAC. Our results indicate that EDC-SAC could be used as a scaffold for regeneration of periodontal tissue. Less
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