BMP2、VEGF165双基因修饰的小鼠BMSCs复合人工骨的体内诱导成骨

In vivo osteogenic potential of calcium phosphate cement seeded with BMP2- and VEGF165-transduced mice BMSCs

目的观察BMP2、VEGF165双基因转染的小鼠BMSCs复合自固化磷酸钙人工骨后的异位诱导成骨能力。方法脂质体介导下将含BMP2、VEGF165双基因的真核表达质粒pIRES转染小鼠BMSCs，转染空白质粒作阴性对照。用免疫组织化学和Western blot方法观察BMP2和VEGF165双基因在小鼠BMSCs内的表达；以自固化磷酸钙人工骨为支架，建立真核表达质粒pIRES—BMP2-VEGF165双基因修饰的组织工程骨植入小鼠右侧股骨肌袋内；将复合转染空白质粒小鼠BMSCs的自固化磷酸钙人工骨植入小鼠左侧股骨肌袋内。于术后第3天、第2，4，6周进行X线摄影对比后处死小鼠，取肌袋内组织行病理切片染色，观察成骨情况。结果小鼠BMSCs在转染BMP2、VEGF165基因后相关蛋白显著表达，由真核表达质粒pIRES—BMP2-VEGF165双基因修饰的组织工程骨在x线和病理学检查中相对于复合转染空白质粒小鼠BMSCs的组织工程骨具有更明显的异位成骨能力。结论由BMP2、VEGF165双基因质粒构建的组织工程骨有明显的异位成骨能力。

Objective To evaluate the ectopic osteogenic potential of calcium phosphate cement seeded with BMP2- and VEGF165-transduced mice BMSCs. Methods BMSCs transfected with recombinant plasmid pIRES-BMP2-VEGF165 using a liposome method were implanted into the muscle pouches of right thigh of the mice. BMSCs transfected only with plRES served as a negative control and implanted into the muscle pouches of right thigh of the mice. Expressions of BMP2 and VEGF165 genes were assayed by immunohistochemical analysis and Western blot. Ectopic bone formation was evaluated by radiography and histology at 3 days and 2, 4, and 6 weeks after implantation. Results Expressions of BMP2 and VEGF165 proteins were highly improved in BMP2- and VEGF165-transduced BMSCs. Calcium phosphate cement with BMSCs transfected with pIRES-BMP2-VEGF165 provided better ectopic osteogenesis compared to control group. Conclusion Tissue-engineered bone with BMP2- and VEGF165-trans- duced mice BMSCs.