聚乙烯亚胺递送miR-218及miR-26a对骨髓间充质干细胞成骨分化的影响  

Effects of miR-218 and miR-26a on osteogenic differentiation of bone marrow mesenchymal stem cells by polyethyleneimine delivery

作　　者：孙璇 魏泽全 布文奂[1] 方滕姣子 西洛片多 刘麒麟[1] 孙宏晨[1] Sun Xuan;Wei Ze-quan;Bu Wen-huan;Fang Tengjiaozi;Xiluopianduo;Liu Qi-lin;Sun Hong-chen(Development and Jaw Remodeling, Hospital of Stomatology, Jilin University, Changchun 130021, Jilin Province, China;Peking University School of Stomatology, Beijing 100034, China)

机构地区：[1]吉林大学口腔医院牙发育及颌骨重塑吉林省重点实验室,吉林省长春市130021 [2]北京大学口腔医学院,北京市100034

出　　处：《中国组织工程研究》2017年第25期3964-3971,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81320108011);吉林省省级产业创新专项资金项目(2016C044-3);吉林省自然科技基金项目(20170101093JC);大学生创新创业训练计划(2015781125)~~

摘　　要：背景:骨髓间充质干细胞因其多向分化潜能广泛应用于组织工程领域,miRNA对于促进骨髓间充质干细胞成骨分化具有重要作用。目的:探究miR-218和miR-26a对大鼠骨髓间充质干细胞成骨分化的影响,为诱导骨髓间充质干细胞成骨分化的相关研究及临床应用提供前期依据和选材参考。方法:提取Wistar大鼠双下肢股骨骨髓,分离培养骨髓间充质干细胞传至第3代备用。miR-218、miR-26a和聚乙烯亚胺(PEI)以特定比例混合孵育形成miRNA/PEI复合物,设立阴性对照组。结果与结论:(1)大鼠骨髓间充质干细胞生长状态良好;(2)MTT法筛选miRNA mimics转染适宜浓度为50 nmol/L;(3)qRT-PCR检测成骨相关基因碱性磷酸酶、Ⅰ型胶原的mR NA表达水平增高(P<0.05);(4)与空白对照组和阴性对照组相比,碱性磷酸酶染色后胞质均产生了较为明显的着色;(5)茜素红染色出现较多矿化结节。结果显示,以PEI为载体构建的miR-218/PEI复合物、miR-26a/PEI复合物及miR-218+miR-26a/PEI共转染复合物对骨髓间充质干细胞的成骨分化具有一定的促进作用,相同条件下miR-26a成骨能力稍优于miR-218。BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are widely used in the field of tissue engineering because of their multi-directional differentiation potential.Micro RNAs play an important role in promoting osteogenic differentiation of BMSCs.OBJECTIVE:To investigate the effect of miR-218and miR-26a on the osteogenic differentiation of rat BMSCs,and to provide reference for the study on osteogenic differentiation of BMSCs and the clinical application.METHODS:The bone marrow of the femur of Wistar rats was extracted and the BMSCs were isolated and cultured to the3rd generation.MiR-218,miR-26a and polyethyleneimine(PEI)were mixed in a specific ratio to form a miRNA/PEI complex.Meanwhile,a negative control group was established.RESULTS AND CONCLUSION:(1)Rat BMSCs grew well.(2)The optimal concentration of miRNA mimics was50nmol/L by MTT method.(3)The expression of alkaline phosphatase and collagen type I mRNA was significantly increased(P<0.05).(4)Alkaline phosphatase staining showed that compared with the blank control group and the negative control group,the cytoplasm showed obvious coloring.(5)There were a lot of mineralized nodules shown by alizarin red staining.Therefore,miR-218/PEI complex,miR-26a/PEI complex and miR-218+miR-26a/PEI co-transfection complex could promote the osteogenic differentiation of BMSCs.Under the same conditions,the osteogenesis of miR-26a was slightly better than that of miR-218.

关 键 词：干细胞 骨髓干细胞 聚乙烯亚胺 MIR-218 miR-26a 骨髓间充质干细胞 成骨分化 RNA干扰 转染 载体 

分 类 号：R394.2[医药卫生—医学遗传学]