Development of new pulp capping/ amputation agent combined with mineral induction activity

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The final goal of this project is to develop new bio - active pulp capping agent using rhBMP-2. First, we examined the effects of rhBMP-2 on differentiation of human pulp cells by measurement of, mRNA expression using quantitative PCR method. We got pulp tissue from extracted fresh wisdom tooth, and obtained pulp cells by out-growth method. We cultured the pulp cells with rhBMP-2 of 0, 10, ; 100 and 1000 ng/ml without serum, and measured various mRNA expressions using LightCycler (Roche, Germany). As a result, rhBMP-2 enhanced dentin sialo phosphoprotein mRNA expression that is thought to be marker of odontoblast differentiation, in a concentration pendent manner. Therefore, we found that rhBMP-2 enhanced the differentiation of pulp cells into odontoblasts, and it is suggested that effective pulp-conservative agent would be produced by using rhBMP-2. Next, We implanted rhBMP-2/ type I collagen/ hydroxyapatite/ phosphophoryn composite into subcutaneous connective tissue or onto skull of rat. Then we sacrificed the rats, and analyzed the implants biochemically and histologically. As a result, we found that rhBMP-2/ type I collagen/ hydroxyapatite/ phosphophoryn composite induced bone formation strongly compared with control group. It was Indicated that rhBMP-2/ type I collagen/ hydroxyapatite/ phosphophoryn composite can be effective pulp capping agent.