Employing Absorbable Coral-hydroxyapatite As A Scaffold Fabricating Tissue Engineering Bone

The regeneration of bone and the repair of bone defects remain a challenging problem in clinical treatment. The emerging and developing of tissue engineering offers tremendous potential for it. The advantages of a tissue engineering approach is that small amounts of cells can harvested, expanded in cell culture, seeded into resorbable matrices, and then used in the repair of large bone tissue defects.In the light of present study , there are still some questions that can't be resolved on the formation and repair of bone: ﹉ow to obtain large amounts of osteoblasts in vitro, ﹉ow to improve the scaffolds to let them support tissue engineering bone formation. (3)how to reconstruct new bone effectively in vitro. Based on these questions, we do some works as follows:1. The biological characteristics of human bone marrow stroma stem cell cultured in vitro: Obtained human fetal bone marrow (hMSCs) and cultivated it immediately in DMEM medium containing lOOmL-L"1 fetal bovine serum. Inducing hMSCs with mineralizational medium for 2-3 weeks. Then the cells were stained by calcifying nodule Von-kossa and Alkaline Phosphatase(ALP) Calcium-Cobalt method. Drew the cell growth curves. Measured the content of ALP and dealt with r-test. The results show that after inducting with mineralizational medium, the growth rate of the cells became slowly. The ability of secreting ALP was enhancing (p< 0.05). And the results of stain reaction were positive. All these showed hMSCs had been differentiated determinately into osteoblasts. hMSCs can be obtained and induced easily. The potential of its clinical application is foreseeing.2.The characteristics of absorbable coral-hydroxyapatite Interpore 500R.Using X-ray diffraction X-ray energy disperse > stereological miscroscope methods to analyze the component and the three-dimensional structure, the degradation rate of InterporeSOOR. The results show Interpore500R is mixture of aragonite and hydroxyapatite. The proportion of calcium carbonate is 90%. The component of InterporeSOOR is different from InterporeSOO and natural coral. Interpore500R is degradable in vivo. In the muscles of New Zealand white rabbit the degradation rate is 50% for six months. The holes of InterporeSOOR are well-distributed and well-communicated. All shows Interpore 500R has ヽontrollable degradation (2) still to remain the components which could promote MSCs differentiation (3) three-dimensional structure advantageous to bone ingrowth. It is an ideal scaffold for bone TE.3.Culturing human bone marrow stromal cells on the surface of Interpore 500R: After inoculating hMSCs onto the surface of Interpore 500R, we surveyed the characteristic of proliferation by cell counting. Using scanning electronic microscope we observed celluar morphology. Staining collagen of I type (immunocytochemistry method) and observing the result under confocal microscope. Measured the content of cellular ALP. The results show hMSCs went on proliferating and started to differentiate to osteoblast after they attched on the material surface. In conclution, the absorbable coral hydroxyapatite is a good scaffold for bone TE.4.1nterpore 500R as scaffold fabricating tissue engineering bone heterotopic in vitro: Induced hMSCs differentiating to osteoblasts. Inoculated hMSCs on absorbable Interpore 500R, then implanted the composite into the muscle of nude mice back. After 60d observed the bone formation using HE dying. It showed the hMSCs /InterporeSOOR composite still remaining the prime volume and shape. The composite could form TE bone. This TE approach is viable. S.The biological characteristics of dog bone marrow stroma stem cell culturedin vitro: The method used is same as part l.In addition we compared dog bone marrow stromal stem cell (dMSCs) to hMSCs. The results showed both dMSCs and hMSCs proliferate quickly. They could be induced to osteoblast. But they still had differences (D dMSCs has strong ability to proliferate.?colony forming unit could be formed in primary dMSCs culture. And calcifying nodule could b...