MiR‐140‐5p promotes osteogenic differentiation of mouse embryonic bone marrow mesenchymal stem cells and post‐fracture healing of mice

Abstract

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 realtime 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. Significance of the study 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 osteogenic medium-treated C3H10T1/2 cells. MiR-140-5p increased the expression of osteocalcin and bone mineral density and bone mass and promoted bone healing of miR-140-5p-transgenic mice with

MicroRNA-140-5p was involved in osteonecrosis of the femoral head by inhibiting osteogenic diffentiation and migration of hBMSCs

“…Previous studies have shown that the development and progression of ONFH are caused by reduced osteogenesis and enhanced adipogenesis of BMSCs 28,29 . Controversial osteogenic effects of MiR-140-5p on MC3T3-E1 cell lines and MSCs have been reported [30][31][32] . In the present study, we found that forced expression of miR-140-5p reduced the expression of collagen type 1and OCN in hBMSCs.…”

The effect of function in mesenchymal stem cells after bilirubin stimulation

“…MSCs can be isolated from autologous and expanded in vitro. As has been widely acknowledged, MSCs can differentiate into several directions, including osteogenic, adipogenic, chondrogenic and neurogenic differentiation [31,32]. We investigated the adipogenic and osteogenic differentiation of MSCs stimulated by bilirubin.…”

Effect of Sperm Cryopreservation on miRNA Expression and Early Embryonic Development

“…Increasing evidences found that many miRNAs associated with the cryopreservation of sperm in mammals are related with fertility, the development of early embryo, and pregnancy. Among the 11 DEmiRs that we identified in human frozen-thawed sperm, hsa-miR-106b-5p ( Wei et al, 2017 ; He et al, 2020 ), hsa-miR-590-3p ( Shi X. et al, 2020 ), hsa-miR-328-3p ( Bai et al, 2021 ), and hsa-miR-140-5p ( Nie et al, 2019 ; Jiao et al, 2020 ) may cause a decreased fertilization rate in IVF and ICSI cases when using cryopreserved sperm since these miRNAs are all related to cell cycle and mitosis. miR-106b-5p is a key miRNA morula and blastocyst developmental state, having potential interactions with embryo-expressed genes ( Sanchez et al, 2021 ).…”

“…Upregulated miR-328-3p is a biomarker in teratozoospermia and may decrease fertilization and embryo formation rate ( Corral-Vazquez et al, 2019 ). Moreover, for the three common DEmiRs between human and mouse sperm, it is reported that overexpression of miR-140-5p significantly inhibits cell proliferation and promotes cell apoptosis in chronic myeloid leukemia cells ( Geng et al, 2020 ; Jiao et al, 2020 ), while miR-30b-5p can promote cell proliferation and cell cycle progression in cancer ( Qin et al, 2017 ), and miR-148b-3p is correlated with fertility rate ( Chen et al, 2018 ; Sekulovski et al, 2021 ). More importantly, in our study, downregulation of miR-148b-3p, according to qRT-PCR analysis, in both human and mouse frozen-thawed sperm may still affect the expression of miR-148b-3p during subsequent embryonic development, leading to the significantly lower embryo formation rate.…”