Supplements for regeneration of dentine-pulp complex by bone marrow cells in porous scaffold

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For regeneration of dentine-pulp complexes, promotion of the hard tissue formation and improvement of the hardness are demanded. Therefore effective supplements for promotion of hard tissue formation and regeneration of dentine-pulp complexes by bone marrow stem cells should be required. Hyaluronic acid, laminin, dextran and amino acid were selected as a supplement for promotion of osteogenesity by differentiated bone marrow stem cells in vitro and in vivo. The results indicated that the osteogenic effect of every chemical substance was inferior to that of dexamethasone by bone marrow cells. However, calcium nodule was deposited conspicuously in bone marrow cell culture by addition of hyaluronic acid, laminin or dextran with dexamethasone in culture medium.Porous HA scaffolds with 55% porosity with and without a hollow center were immersed in each chemical substance and air dried respectively. They were seeded of bone marrow stem cells. After 4 weeks of dorsal subcutaneous implantation in vivo, bone formation in many pores was observed histologically. In each hyaluronic acid, laminin or dextran processed scaffold with seeding of only 1×10^6 cells/ml concentration of bone marrow cells, bone formation in remarkable number of the pores was recognized. Bone formation was confirmed by immunochemical quantitative analysis of osteocalcin in the scaffolds. It was suggested that an effect of these chemical substances to promote bone formation effectively by addition in the bone marrow cell culture medium with dexamethasone. Hyaluronic acid, laminin or dextran induced many bone formation in a pore by a small number of cells. It was because of cell adhesion property of the agents to the inner wall of pores of the scaffold. Moreover, the nutrition supply via a hollow center might be sufficient for bone marrow cell proliferation and differentiation.