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Construction of Tissue Engineering Bone with Osteoactivin Gene Transfected Rabbit Bone Marrow Stromal Cells and Porous Silk Fibroin Scaffold in Vitro
王涵, 杨征毅, 程峰, 潘广嗣, 易晓辉, 孙晋, 曹依娜, 袁林
骨活素, 骨髓基质干细胞, 基因治疗, 组织工程骨Osteoactivin, Bone Marrow Stromal Cells, Gene Therapy, Tissue Engineering Bone
《Advances in Clinical Medicine》, Vol.4 No.2, 2014-06-23
Objective: Rabbit bone marrow stromal cells (BMSCs) infected by a recombinant adenoviral vector carrying the osteoactivin gene (Ad-OA) were seeded into porous silk fibroin scaffold to construct tissue engineering bone in vitro. Method: Ad-OA infected RBM SC cultured in vitro and the expression of OA in these cells after infection were determined by in situ hybridization and immune ohistochemical analysis. OA productions were confirmed by western blot analysis of the supernatant collected from the cells. The changes of cellular proliferation and differentiation in the cells were observed by flowcytometry and ALP activity analysis. OA transduced cells were then seeded into porous silk fibroin scaffolds. The attachment and growth of the cells on the scaffold were examined using SEM. Results: The expression of OA was confirmed in mRNA and protein levels in the cells after infection and the presence of OA was detected in the supernatant of the cells. In addition, cellular proportion in S period and ALP activity obviously increased in the cells. SEM examination revealed extensive cellular attachment and growth on the porous silk fibroin scaffolds composite in 1 day. Conclusion: Ad-OA could infect RBMSC with high efficiency and promote cellular proliferation and osteoblast conversion. The cells after infection grew well on a porous silk fibroin scaffold. Tissue engineering bone used to regional gene therapy is constructed successfully.