转染BMP-2基因的兔BMSCs种植PLA/PCL支架体外构建组织工程骨

Bone marrow stromal cells carrying human bone morphogenetic protein-2 gene seeded into PLA/PCL scaffold to construct tissue engineering bone in vitro

目的 :用腺病毒载体将人骨形态发生蛋白 2 (BMP 2 )基因导入兔骨髓基质干细胞 (BMSCs) ,种植PLA/PCL (聚乳酸 /聚己内酯 )支架体外构建组织工程骨。方法 :蛋白印迹法检测转染后细胞BMP 2的表达 ,流式细胞仪和ALP活性检测分析基因转染对细胞增殖分化的影响。然后将转染后细胞接种到PLA/PCL支架上 ,扫描电镜观察细胞贴附、生长状况。结果 :转染后 ,BMSCs表达BMP 2 ,S期细胞比例和ALP活性明显增高。扫描电镜见转染细胞分布均匀 ,伸展良好。结论 :BMP 2基因转染BMSCs ,可促进细胞增殖分化。转染后细胞在PLA/PCL支架上生长良好 ,BMP 2基因治疗的组织工程骨构建成功。

Objective:Rabbit bone marrow stromal cells (BMSCs) infected by a recombinant adenoviral vector carrying the human BMP 2 gene (Ad BMP 2) were seeded into PLA/PCL scaffolds to construct tissue engineering bone in vitro Method:The expression of hBMP 2 in these cells after infection was determined by Western blot analysis The changes of cellular proliferation and differentiation were observed by flow cytometer and ALP activity analysis BMP 2 transduced cells were then seeded into PLA/PCL scaffolds The attachment and growth of the cells on the scaffold were examined using SEM Result:The expression of hBMP 2 was confirmed and ALP activity obviously increased in the cells after infection SEM examination revealed extensive cellular attachment and growth on the PLA/PCL block Conclusion:Ad BMP 2 could infect BMSCs with high efficiency and promote its diferentiation The cells after infection grow well on PLA/PCL scaffold Tissue enginerring bone used to regional gene therapy is constructed successfully