MicroRNA-126慢病毒增强MSCs-水凝胶修复下颌骨缺损效果的研究

摘要

Objective To study the effect of lentiviral vector of MicroRNA-126 on survival differentiation of mesenchymal stem cells under ischemic condition.Methods Lentiviral vector of MicroRNA-126 was constructed and then transfected into mouse mesenchymal stem cells (MSCs).Flow detection was used to detect transfection ability and synthesis of collagen type Ⅰ.ELISA was used to detect osteocalcin,Insulin-like growth factor 1 (IGF-1) and transforming growth factor-β (TGF-β) in cell culture supernatant.Mandibular bone defect model was constructed and then injected the hydrogel mixed with MSCs.After 48h,the survival rate and proportion of differentiation into osteoblasts was determined.Results Flow results showed lentiviral vector of MicroRNA-126 could transfected into MSCs.ELISA results showed that MicroRNA-126 lentiviral vector transfected MSCs secreted more bone gla protein and IGF-1 and TGF-β,which increased by 1.43,1.87 and 2.63 times compared with the control virus treated group.Animal experiments showed the number of stem cells survived in the MicroRNA-126 group was 4.56 times that of the control group and the percentage of MSC differentiation into osteoblasts was 75％,which is 3.95 times more than that in the control group.Conclusion The research shows that lentiviral vector of MicroRNA-126 could maintain MSCs survival and differentiation under ischemic condition.%目的 构建MicroRNA-126慢病毒载体以及研究MicroRNA-126对缺血条件下间充质干细胞存活和成骨分化的影响.方法 构建MicroRNA-126慢病毒载体并感染间充质干细胞(Mesenchymal stem cells,MSCs);流式检测转染能力和MSC合成Ⅰ型胶原能力;ELISA检测细胞培养上清中骨钙素、胰岛素样生长因子1(Insulin-like growth factor 1,IGF-1)和转化生长因子β(Transforming growth factor-β,TGF-β)的含量;构建下颌骨骨缺损模型,在体内注射混合有MSCs的水凝胶,移植48 h,检测存活率和分化为成骨细胞的比率.结果 流式的结果显示MicroRNA-126的慢病毒载体能顺利的转染进入细胞;ELISA的结果显示MicroRNA-126的慢病毒载体转染的MSC能分泌更多的骨钙素、IGF-1和TGF-β,相对于对照病毒处理组分别上升了1.43、1.87和2.63倍(P＜0.05);动物实验的结果显示移植48h后,MicroRNA-126组存活的干细胞的数量是对照病毒组的4.56倍,分化为成骨细胞的MSC的比例为75％,是对照病毒组的3.95倍(P＜0.05).结论 MicroRNA-126可以维持缺血条件下间充质干细胞存活及其向成骨细胞的分化.