Regeneration of tooth supporting tissue by the implantation of cell fixed collagen membrane

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The aim of this study is to promote the regeneration of tooth supporting tissue by the implantation of'cell fixed collagen membrane. The crucial stage in this study is creation of cell fixed membrane, and I first investigated features of cells to satisfy the regeneration of tooth supporting tissue by the collagen membrane. The stem cells harvested from bone marrow were fixed in the collagen membrane, and the membrane was cultured for four weeks to analyze bone forming activity. The bone forming activity was estimated by the measurement of alkaline phosphatase activity and detection of osteoblast-specific gene (osteocalcin, bone sialoprotein), and I recognized bone forming activity in the collagen membrane. To promote the bone forming activity, Bone Morphogenetic Protein (BMP), of basic Fibroblast Growth Factor (bFGF) was added in the membrane. When BMP was mixed in the collagen membrane, the bone forming activity appeared faster. However, the intensity of bone forming activity did not change by the addition of BMP. This might be due to the enhancement of osteoblastio differentiation by BMP. Next I examined the effect of bFGF. The total bone forming activity of the membrane increased when high dose of bFGF was mixed in the collagen membrane. However the bone forming activity of each cell decreased. This finding might indicate that bFGF accelerated cell growth, but suppressed the expression of osteoblastic phenotype. When low dose of bFGF was mixed in the collagen membrane, the bone forming activity of membrane increased. However this activation did not occurred if bFGF was not added in the culture medium. This might be due to the loss of bFGFfrom the collagen membrane. I am going to create the new cell fixed membrane which could maintain the low dose of bFGF for a long time.