MiR‐140‐5p is high expressed in normal fracture healing, but its specific role and mechanism in tissue‐to‐bone healing are rarely reported. Therefore, this study investigated the effects of miR‐140‐5p on tissue‐to‐bone healing. Clone formation experiment, flow cytometry, Alizarin Red S Staining and Oil Red O Staining were performed to investigate the biological characteristics of mouse embryonic bone marrow mesenchymal stem cells C3H10T1/2. MiR‐140‐5p mimic was transfected into osteogenic medium (OS)‐treated C3H10T1/2 cells to investigate the effects of miR‐140‐5p on osteogenic differentiation. MiR‐140‐5p transgenic mouse model and the transgenic fracture model were established, and the effects of miR‐140‐5p on osteogenic differentiation, bone mineral density (BMD) and bone mass of bone tissues were detected by haematoxylin and eosin staining and computed tomography scan. The expressions of osteocalcin, differentiation‐related genes (Runx2, ALP, Spp1 and Bglap3) and miR‐140‐5p were determined by quantitative real‐time polymerase chain reaction. C3H10T1/2 cells showed the abilities of forming cloned differentiation of osteogenesis, fat cells, and its phenotypes including CD44, CD90.1 and Sca‐1 but excluding CD45 haematopoietic stem cell marker. Overexpression of miR‐140‐5p promoted the expressions of differentiation‐related genes and calcium deposition of OS‐treated C3H10T1/2 cells. MiR‐140‐5p increased the expression of osteocalcin, BMD and bone mass and promoted bone healing of miR‐140‐5p‐transgenic mice with fracture. MiR‐140‐5p promoted osteogenic differentiation of mouse embryonic bone marrow mesenchymal stem cells and post‐fracture healing in mice.

中文翻译：

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MiR‐140‐5p促进小鼠胚胎骨髓间充质干细胞的成骨分化和小鼠骨折后的愈合

MiR140-5p在正常骨折愈合中高表达，但很少报道其在组织至骨愈合中的特定作用和机制。因此，本研究调查了miR‐140‐5p对组织至骨愈合的影响。进行了克隆形成实验，流式细胞术，茜素红S染色和油红O染色研究小鼠胚胎骨髓间充质干细胞C3H10T1 / 2的生物学特性。将MiR‐140‐5p模拟物转染到成骨培养基（OS）处理的C3H10T1 / 2细胞中，以研究miR‐140‐5p对成骨细胞分化的影响。建立了MiR‐140‐5p转基因小鼠模型和转基因骨折模型，并且miR‐140‐5p对成骨细胞分化的影响，用苏木精和曙红染色及计算机断层扫描检查骨密度和骨组织的骨量。通过定量实时聚合酶链反应测定骨钙蛋白，分化相关基因（Runx2，ALP，Spp1和Bglap3）和miR-140-5p的表达。C3H10T1 / 2细胞具有形成成骨，脂肪细胞及其表型（包括CD44，CD90.1和Sca-1）的克隆分化能力，但不包括CD45造血干细胞标记。miR-140-5p的过表达促进了OS处理的C3H10T1 / 2细胞的分化相关基因的表达和钙沉积。MiR‐140‐5p可增加miR‐140‐5p‐转基因骨折小鼠的骨钙素，BMD和骨量的表达，并促进骨愈合。