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
发布时间：2018-09-10 17:55
【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(P0.05);(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。
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) are widely used in tissue engineering for their multidirectional differentiation potential. MiRNA plays an important role in promoting osteogenic differentiation of BMSCs. Objective: to investigate the effects of miR-218 and miR-26a on osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to provide a preliminary basis and reference for clinical application in inducing osteogenic differentiation of BMSCs. Methods: bone marrow of both lower extremities of Wistar rats were isolated and cultured. Bone marrow mesenchymal stem cells were isolated and transferred to the third generation. MiR-218miR-26a and polyethyleneimide (PEI) were incubated in a specific proportion to form miRNA/PEI complex, and negative control group was set up. Results and conclusion: (1) Rat bone marrow mesenchymal stem cells (BMSCs) grew well; (2) the suitable concentration of miRNA mimics transfection was 50 nmol/L; (3) qRT-PCR to detect alkaline phosphatase (ALP), which was associated with osteogenesis. Compared with the control group and the negative control group, the mR NA expression level of type I collagen increased significantly (P0.05); (4), the cytoplasm of alkaline phosphatase staining was significantly stained; (5) alizarin red staining showed more mineralized nodules. The results showed that the miR-218/PEI complex constructed with PEI as the carrier could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and the osteogenic ability of miR-26a was slightly better than that of miR-218. under the same conditions.
【作者单位】： 吉林大学口腔医院牙发育及颌骨重塑吉林省重点实验室;北京大学口腔医学院;
【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)~~
【分类号】：R68
，
本文编号：2235184
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) are widely used in tissue engineering for their multidirectional differentiation potential. MiRNA plays an important role in promoting osteogenic differentiation of BMSCs. Objective: to investigate the effects of miR-218 and miR-26a on osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to provide a preliminary basis and reference for clinical application in inducing osteogenic differentiation of BMSCs. Methods: bone marrow of both lower extremities of Wistar rats were isolated and cultured. Bone marrow mesenchymal stem cells were isolated and transferred to the third generation. MiR-218miR-26a and polyethyleneimide (PEI) were incubated in a specific proportion to form miRNA/PEI complex, and negative control group was set up. Results and conclusion: (1) Rat bone marrow mesenchymal stem cells (BMSCs) grew well; (2) the suitable concentration of miRNA mimics transfection was 50 nmol/L; (3) qRT-PCR to detect alkaline phosphatase (ALP), which was associated with osteogenesis. Compared with the control group and the negative control group, the mR NA expression level of type I collagen increased significantly (P0.05); (4), the cytoplasm of alkaline phosphatase staining was significantly stained; (5) alizarin red staining showed more mineralized nodules. The results showed that the miR-218/PEI complex constructed with PEI as the carrier could promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and the osteogenic ability of miR-26a was slightly better than that of miR-218. under the same conditions.
【作者单位】： 吉林大学口腔医院牙发育及颌骨重塑吉林省重点实验室;北京大学口腔医学院;
【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)~~
【分类号】：R68
，
本文编号：2235184
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