REPAIR OF CRITICAL-SIZED BONE DEFECTS WITH ANTI-MIR-31-EXPRESSING BONE MARROW STROMAL STEM CELLS AND POLY(GLYCEROL SEBACATE) SCAFFOLDS
REPAIR OF CRITICAL-SIZED BONE DEFECTS WITH ANTI-MIR-31-EXPRESSING BONE MARROW STROMAL STEM CELLS AND POLY(GLYCEROL SEBACATE) SCAFFOLDS
复制标题
使用表达抗 miR-31 的骨髓基质干细胞和聚（癸二酸甘油酯）支架修复临界尺寸的骨缺损。
DOI：
10.22203/ecm.v027a02
复制
发表时间：
2014-01-01
影响因子：
3.1
通讯作者：
Fan, Xianqun
中科院分区：
文献类型：
--
作者：
Deng, Yuan;Bi, Xiaoping;Fan, Xianqun
骨组织工程中关键尺寸缺损的修复是一个重大挑战。结合使用祖细胞与基因治疗代表了一种有前途的方法骨再生。microRNA在大多数基因调控网络中发挥重要作用，调节多种生长因子的内源性表达，同时调节干细胞分化。我们前期的研究表明，敲低miR-31可促进骨髓基质干细胞（BMSCs）的成骨作用。为了研究工程化以表达用于CSD修复的抗miR-31的细胞的治疗潜力，构建编码阴性对照、miR-31前体和反义序列的慢病毒载体，并转导到骨诱导BMSC中。检测成骨特异性基因表达、碱性磷酸酶活性和茜素红S染色，评价miR-31对BMSCs 3周内细胞命运的影响。此外，将miR-31修饰的BMSC接种在聚（甘油癸二酸酯）（PGS）支架上用于修复大鼠中8 mm临界尺寸的颅骨缺损。结果显示，miR-31抑制在mRNA和蛋白水平上显著增加体外成骨特异性基因的表达，并且在体内抗miR组中观察到具有高局部骨矿物质密度的稳健的新骨形成。此外，携带表达抗miR-31的BMSCs的PGS支架在体内骨缺损内表现出良好的生物相容性和高再生率（约60%）。我们的研究结果表明，miR-31基因的传递影响BMSCs的成骨分化和骨再生的潜力，PGS是一个潜在的基板，基因修饰，组织工程骨修复大骨缺损。
The repair of critical-sized defects (CSDs) is a significant challenge in bone tissue engineering. Combining the use of progenitor cells with gene therapy represents a promising approach for bone regeneration. MicroRNAs play important roles in most gene regulatory networks, regulate the endogenous expression of multiple growth factors and simultaneously modulate stem cell differentiation. Our previous study showed that knocking down miR-31 promotes the osteogenesis of bone marrow stromal stem cells (BMSCs). To investigate the therapeutic potential of cells engineered to express anti-miR-31 for CSD repair, lentiviral vectors encoding negative control, miR-31 precursor and anti-sense sequences were constructed and transduced into osteo-inductive BMSCs. The expression of osteogenic-specific genes, alkaline phosphatase activity and Alizarin Red S staining were investigated to evaluate the effects of miR-31 on the cell fate of BMSCs over a 3-week period. In addition, miR-31-modified BMSCs seeded on poly(glycerol sebacate) (PGS) scaffolds were used to repair 8 mm critical-sized calvarial defects in rats. The results showed that miR-31 suppression significantly increased the expression of osteogenic-specific genes in vitro at the mRNA and protein levels, and that robust new bone formation with high local bone mineral density was observed in the anti-miR groups in vivo. Moreover, the PGS scaffolds carrying anti-miR-31-expressing BMSCs exhibited good biocompatibility and a high regeneration rate (~60%) within in vivo bone defects. Our results suggest that miR-31 gene delivery affects the potential of BMSCs for osteogenic differentiation and bone regeneration and that PGS is a potential substrate for genetically modified, tissue-engineered bone in the repair of large bone defects.