不同诱导阶段的骨髓间充质干细胞修复兔骨损伤的实验研究

The Efficacy of Different Stages of Induced BMSCs in Repairing Bone Injury of Rabbit

目的对比不同诱导阶段的骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)对兔骨折损伤的疗效,并找到适宜移植的最佳诱导时间。方法体外分离培养BMSCs,表型鉴定。绿色荧光蛋白(Green fluorescent protein,GFP)转基因标记后,定向诱导。将实验兔40只随机分为5组:未诱导组5只;诱导3 d组10只;诱导7 d组10只;诱导21 d组10只;对照组5只。建立兔尺骨骨折动物模型,造模24 h后,将GFP标记的不同诱导阶段细胞按3×106cells/Kg于骨折处局部注射,移植后行X线检查并评分。结果骨折处X线评分,BMSCs未诱导组、诱导7 d、诱导21 d组评分比较无明显差异(P>0.05),且均显著高于对照组(P<0.01),3 d组评分高于7 d和21 d组(P<0.05),且显著高于对照组(P<0.01)。结论BMSCs成骨诱导3 d可能是最适宜移植的。

Objective To compare the efficacy of different stages of induced BMSCs in repairing bone injury of rabbit, and to find out the most suitable cells for transplantation. Methods BMSCs were isolated, cultured and identified in vitro. After labeled by GFP, BMSCs were induced and 40 rabbits were randomly divided into five groups according to different time of induction： non-induce group （n=5）, 3-day group （n=10）, 7-day group （n=10）, 21-day group （n=10）, control group （n=5）. The bone injury model of rabbit was established. After successfully modeled for 24 hours, GFP labeled, induced BMSCs were injected into fracture area with the dosage of 3×106 cells/Kg. X-ray exam was carried out and evaluated. Results According to X-ray evaluation criteria, the X-ray score among non-induce group, 7-day group and 21-day group has no difference （P〉0.05）, but all significantly higher than control group （P〈0.01）. The X-ray score in 3-day group was higher than 7-day group and 21-day group （P〈0.05）, and significantly higher than control group （P〈0.01）. Conclusion BMSCs induced for 3 days turned out to be the most suitable cells for transplantation.