The role of miR-31-modified adipose tissue-derived stem cells in repairing rat critical-sized calvarial defects
The role of miR-31-modified adipose tissue-derived stem cells in repairing rat critical-sized calvarial defects
复制标题
miR-31 修饰的脂肪组织来源的干细胞在修复大鼠临界大小的颅骨缺损中的作用。
DOI：
10.1016/j.biomaterials.2013.05.042
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发表时间：
2013-09-01
期刊：
影响因子：
14
通讯作者：
Fan, Xianqun
中科院分区：
文献类型：
--
作者：
Deng, Yuan;Zhou, Huifang;Fan, Xianqun
随着microRNAs（miRNAs）在各种疾病的治疗和监测中的应用日益广泛，miRNAs已成为生物学和医学研究的重要工具。最近的研究表明，miRNAs参与了干细胞的成骨分化。然而，很少有研究报道使用miRNA修饰的成体干细胞使用组织工程技术修复临界尺寸缺陷（CSD）。已知miR-31是一种多效作用的miRNA，其抑制癌症转移并靶向成纤维细胞中的特殊富含AT的序列结合蛋白2（Satb 2）。然而，目前尚不清楚miR-31的功能是否是在成骨分化和骨再生期间增强脂肪组织来源的干细胞（ASC）成骨，沿着其与Satb 2的关联。在这项研究中，我们系统地评估了miR-31在增强ASC成骨中的功能以及miR-31修饰的ASC在大鼠CSD模型中的治疗潜力，该模型采用β-磷酸三钙（β-TCP）支架。用慢病毒（Lenti）-miR-31、Lenti-as-miR-31（反义）或Lenti-NC（阴性对照）处理ASC。然后将这些基因修饰的ASCs与β-TCP支架组合以修复大鼠的CSD。结果表明，Lenti-as-miR-31可显著增强体外培养的ASCs成骨mRNA和蛋白的表达。此外，我们首次发现骨形态发生蛋白-2（BMP-2）启动的Runt相关转录因子2（Runx 2）、Satb 2和miR-31调控环在ASCs的成骨分化和骨再生中发挥重要作用。更重要的是，我们发现miR-31敲低的ASCs显着改善了CSD的修复，包括增加骨体积，增加骨矿物质密度（BMD）和减少体内支架残留。这些数据证实了miR-31修饰的ASC在体外和体内成骨中的重要作用。(C)2013爱思唯尔有限公司版权所有。
With the increasing application of microRNAs (miRNAs) in the treatment and monitoring of different diseases, miRNAs have become an important tool in biological and medical research. Recent studies have proven that miRNAs are involved in the osteogenic differentiation of stem cells. However, few studies have reported the use of miRNA-modified adult stem cells to repair critical-sized defects (CSDs) using tissue engineering technology. It is known that miR-31 is a pleiotropically acting miRNA that inhibits cancer metastasis and targets special AT-rich sequence-binding protein 2 (Satb2) in fibroblasts. However, it is not clear whether the function of miR-31 is to enhance adipose tissue-derived stem cell (ASC) osteogenesis, along with its association with Satb2, during osteogenic differentiation and bone regeneration. In this study, we systematically evaluated the function of miR-31 in enhancing ASC osteogenesis and the therapeutic potential of miR-31-modified ASCs in a rat CSD model with beta-tricalcium phosphate (beta-TCP) scaffolds. ASCs were treated with lentivirus (Lenti)-miR-31, Lenti-as-miR-31 (antisense) or Lenti-NC (negative control). These genetically modified ASCs were then combined with beta-TCP scaffolds to repair CSDs in rats. The results showed that in cultured ASCs in vitro, Lenti-as-miR-31 significantly enhanced osteogenic mRNA and protein expression when compared with the Lenti-NC group. Moreover, we firstly found that a Runt-related transcription factor 2 (Runx2), Satb2 and miR-31 regulatory loop triggered by bone morphogenetic protein-2 (BMP-2) plays an important role in ASCs' osteogenic differentiation and bone regeneration. More importantly, we found that miR-31-knockdown ASCs dramatically improved the repair of CSDs, including increased bone volume, increased bone mineral density (BMD) and decreased scaffold residue in vivo. These data confirm the essential role of miR-31-modified ASCs in osteogenesis in vitro and in vivo. (C) 2013 Elsevier Ltd. All rights reserved.