论文部分内容阅读
目的观察重组人骨形成蛋白4(human bone morphogenetic protein4,hBMP4)基因腺相关病毒载体诱导兔骨髓间充质干细胞(BMSCs)成骨方向分化的作用。方法全骨髓法培养兔BMSCs,应用重组人骨形成蛋白4基因腺相关病毒MOI值为5×104vg/cell转染兔BMSCs,观察细胞形态,行ALP染色、Von Kossa染色、茜素红染色及ALP含量测定,观察成骨活性。结果AAV-hBMP4转染BMSCs后,细胞形态呈现典型的成骨改变,ALP染色及Von Kossa、染色茜素红染色均出现成骨的特征性改变。细胞上清ALP含量测定,实验组ALP含量明显增高(p<0.01)。结论实验获得的病毒载体滴度高、感染性好,完全可以满足骨组织工程的需要。AAV-hBMP4转染效率高,AAV-hBMP4转染BMSCs后,BMSCs表现明显的成骨活性改变。
Objective To observe the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs) induced by adeno-associated virus vector of recombinant human bone morphogenetic protein 4 (hBMP4). Methods Rabbit BMSCs were cultured by whole bone marrow transplantation. Rabbit BMSCs were transfected with 5 × 104 vg / cell MOI of recombinant human bone morphogenetic protein 4 gene. ALP staining, Von Kossa staining, alizarin red staining and ALP content Measurement, observation of osteogenic activity. RESULTS: After transfection of AAV-hBMP4 into BMSCs, the morphology of the cells was characterized by typical osteoblastic changes. ALP staining, Von Kossa staining and alizarin red staining showed characteristic changes in osteogenesis. The content of ALP in the cell supernatant was measured, and the ALP content in the experimental group was significantly increased (p <0.01). Conclusion The experiment results show that the viral vector has high titer and good infectivity, which can fully meet the needs of bone tissue engineering. AAV-hBMP4 transfection efficiency, AAV-hBMP4 transfected BMSCs, BMSCs showed significant changes in osteogenic activity.