Tissue Engineered Bone Constructed By Seeding Rabbit Marrow Stromal Cell On Porous β-TCP Bioceramic And Coral

Part ⅠEstrogen enhance proliferation and differentiation of in rabbit bone marrow stromal cells culture Object :The study tested the ability of estrogen to regulate growth and differentiation in rabbit bone marrow stromal cells in vitro and provide the suitable seed-cells for bone tissueengineering. Methods: MSCs of the bone marrow of the rabbits were isolated by density gradient sepration method and cultured, then MSCs at the third passage were cultured in subculture incuding estrogen, vitamin C and β-glycerophosphate, the proliferation capability and osteogenic differentiation were investgated .Results: MSCs were spindle-shaped and have strong capability of proliferation.Alkaline phosphates activity of MSCs at third passage were increased in conditional medium and the mineral nodules were observed. Conclusion: MSCs have a strong activity of proliferation and osteogenic potential. Estrogen enhances MSCs of rabbit proliferation, but also stimulate it differentiation. Part ⅡComparative study of the biocompatibility of Coral and porous β-TCP bioceramic Object : To study the the biocompatibility of Coral and porousβ-TCP bioceramic in vitro and in vivo . Methods: MSCs were cultured together with coral and porous β-TCP bioceramic in vitro, induced by estrogen, vitamin C and β-glycerophosphate, then cell /porousβ-TCP bioceramic and cell /coral composites were implanted subcutaneously into rabbits.In vitro morphological characters, cell proliferation ,alkaline phosphatase activity were detected; in vivo the effectiveness of bone formation was evaluated by means of gross, histological observation. Results: In vitro cell can grow on two kinds of material, but on coral little cells died. The numbers of cells increases in every groups, after cultured 12days, the numbers of cells on porous β-TCP bioceramic was greater than on coral, there was statistically significant difference .The contration of ALP in porousβ-TCP bioceramic group was lower than in coral group, there was no statistically significant difference. In vivo, bone were observed in two groups, but there was little cell of inflammation in coral group. At 12 weeks after operation, porousβ-TCP bioceramic has been degenerated, but there was a little coral. Conclusion: In vitro and in vivo, porousβ-TCP bioceramic has better biocompatibility than coral. Part ⅢComparative study of the repair of radius bone defect with porous β-TCP bioceramic and Coral Object : To study the the repair of radius bone defect with porous β-TCP bioceramic and Coral . Methods: Bone defects for 10mm were created on left radius of 48 adult white New Zealand rabbits,then were randomly divided into group ABC.In the group A,management A (MSCs+ porous β-TCP bioceramic) was given to the defect of one radius. In the group B, management B (MSCs+ Coral) was given. In the group C, management C (control) was given. The effect was observed by gross, X-ray, histopathological examinations 4,8,12,20 weeks after operation. Results: Gross and histopathological examinations, in group A and B,the defect were repaired by bone; in group C, the defect were filled with fibrous tisues. In group A, there was little cell of inflammation, porous β-TCP bioceramic was degenerated at 12weeks after implation. In group B, there was a little cells of inflammation, coral was degenerated at 20 weeks after operation. X-ray examinations, 20 weeks after operation, in the management A all speciments had complete bony union; in management B there were 2 speciments of ununion; in management C all are ununion. Conclusion: The compoisitte of MSCs and porous β-TCP bioceramic is better effective repair of radius bone defect than the compoisitte of MSCs and coral.