[P-086] Combination of hPDLSC sheet and rhBMP-2 for periodontal tissue regeneration
研修コード：2504
Keywords:Periodontal tissue、Tissue regeneration、Stem cells、Tissue engineering
Objectives: This study evaluates the efficacy of rhBMP-2 combined with human periodontal ligament stem cells （hPDLSCs） sheet for regeneration of cementum and periodontal ligament.
Materials and methods: To test if rhBMP-2 affects mineralization of hPDLSCs in vitro, monoclonal hPDLSCs were cultured under mineralization condition with or without rhBMP-2 for 21 days. The amount of mineralization was measured by Alizarin Red staining and osteo/cemento-genic gene expressions were analyzed by RT-PCR. To test efficacy of rhBMP-2 on cementum formation in vivo, hPDLSC sheets were fabricated and differentiated with or without rhBMP-2 ex vivo, wrapped around a scaffold disc and transplanted into nude mice. Hard tissue formation was analyzed at 4 weeks after transplantation by H&E staining and immunohistochemistry. Viability of transplanted hPDLSC was evaluated by luciferase in vivo imaging.
Results: rhBMP-2 treatment enhanced mineralization differentiation of hPDLSCs in vitro and the amount increased with longer rhBMP-2 treatment. Expressions of ALP, osteocalcin and cemp-1 were higher in rhBMP-2 group especially at 2 weeks of differentiation. The amount of hard tissue formation in vivo was significantly higher in rhBMP-2 group compared to cell sheet only group. Importantly, the regenerated hard tissue in rhBMP-2 group expressed CEMP-1. Viability of the cell sheet lasted for 3 weeks of transplantation.
Conclusions: Combined usage of rhBMP-2 with hPDLSC sheet results in superior formation of cementum-like tissue in vivo. Our findings have implications for clinical usage of rhBMP-2 with hPDLSC sheet for periodontal regeneration.
This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Instiute （KHIDI）, funded by the Ministry of Health & Welfare, Republic of Korea （grant number : HI17C0450）.
Materials and methods: To test if rhBMP-2 affects mineralization of hPDLSCs in vitro, monoclonal hPDLSCs were cultured under mineralization condition with or without rhBMP-2 for 21 days. The amount of mineralization was measured by Alizarin Red staining and osteo/cemento-genic gene expressions were analyzed by RT-PCR. To test efficacy of rhBMP-2 on cementum formation in vivo, hPDLSC sheets were fabricated and differentiated with or without rhBMP-2 ex vivo, wrapped around a scaffold disc and transplanted into nude mice. Hard tissue formation was analyzed at 4 weeks after transplantation by H&E staining and immunohistochemistry. Viability of transplanted hPDLSC was evaluated by luciferase in vivo imaging.
Results: rhBMP-2 treatment enhanced mineralization differentiation of hPDLSCs in vitro and the amount increased with longer rhBMP-2 treatment. Expressions of ALP, osteocalcin and cemp-1 were higher in rhBMP-2 group especially at 2 weeks of differentiation. The amount of hard tissue formation in vivo was significantly higher in rhBMP-2 group compared to cell sheet only group. Importantly, the regenerated hard tissue in rhBMP-2 group expressed CEMP-1. Viability of the cell sheet lasted for 3 weeks of transplantation.
Conclusions: Combined usage of rhBMP-2 with hPDLSC sheet results in superior formation of cementum-like tissue in vivo. Our findings have implications for clinical usage of rhBMP-2 with hPDLSC sheet for periodontal regeneration.
This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Instiute （KHIDI）, funded by the Ministry of Health & Welfare, Republic of Korea （grant number : HI17C0450）.