骨形态发生蛋白-2基因转染骨髓间充质干细胞移植对糖尿病大鼠骨折愈合的影响

Effects of transplanting bone marrow stem cells transfected with bone morphogenetic protein-2 onfracture healing in diabetic rats

目的观察骨形态发生蛋白-2（BMP-2）基因转染大鼠骨髓间充质干细胞（BM-SCs）移植对糖尿病骨折愈合的影响，探讨糖尿病骨折治疗的新方法。方法Wistar雄性成年6周龄大鼠50只，按随机数字表法分为对照组和实验组，均制成糖尿病骨折模型，体外高糖环境下腺病毒介导的BMP-2转染BMSCs，分别于骨折局部移植相应的BMSCs。术后1，2，3，4，6周行X线检查；取骨痂行HE染色，免疫组化检测骨痂中BMP-2的灰度值；ELISA法检测血清中BMP-2的浓度。结果第4周实验组骨痂BMP-2灰度值为83±3，对照组为1184-4（P〈0．01）。第4周实验组血清BMP-2浓度为（203．80±8．96）ng／L，对照组为（139．15±4．19）ng／L（P〈0．01）。结论BMP-2基因转染的BMSCs移植能促进糖尿病大鼠骨折的愈合。

Objective To observe the effects of transplantation of bone marrow stem cells （BM- SCs） transfected with bone morphogenetic protein-2 （BMP-2） on fracture healing in rats with diabetes so as to provide a new therapy for diabetic fractures. Methods Fifty male adult Wistar rats aged six weeks randomized to the control group and experimental group were all employed to establish models with diabetic fractures. Under high glucose condition, BMSCs were transfected with BMP-2 by adenovirus vec- tor in vitro. BMSCs transfected by BMP-2 were transplanted into the fracture area of rats in the experi- mental group, while non-transfected BMSCs into the corresponding area of rats in the control group. X-ray examination was performed at 1, 2, 3, 4 and 6 weeks after transplantation. Bony calluses were collected for HE staining and gray scales of BMP-2 in calluses were determined by immunohistochemical method. Meanwhile, serum levels of BMP-2 were measured by ELISA. Results The gray scales of BMP-2 in the calluses were 83 ±3 in the experimental group and 118 ±4 in the control group at four weeks （P 〈 0.01 ）. The serum concentrations of BMP-2 were （203.80 ± 8.96） ng/L in the experimental group and （ 139.15 ± 4.19） ng/L in the control group at four weeks （P 〈 0.01 ）. Conclusion Transplantation of BMSCs transfected by BMP-2 promotes fracture healing in diabetic rats.