MicroRNA‐98 regulates osteogenic differentiation of human bone mesenchymal stromal cells by targeting BMP2

Abstract

To study the effects of microRNA‐98 (miR‐98) on human bone mesenchymal stromal cells (hBMSCs). The patients undergoing hip arthroplasty were selected by inclusion/exclusion criteria for this study. The extracted hBMSCs were detected of osteogenic differentiation by alizarin red S staining, and of cell phenotype by flow cytometry. Bioinformatics, dual luciferase report, western blotting, RT‐PCR and immunoblotting were used in our study. The hBMSCs were divided into miR‐98 mimics, miR‐98 negative control (NC), miR‐98 inhibitors, Mock and miR‐98 inhibitors + siBMP2 groups. Human bone mesenchymal stromal cells were extracted and purified in vitro and had specific cytological morphology, surface markers and abilities of self‐renewal and differentiation. Compared with the NC group and Mock group, the miR‐98 mimics group showed increased miR‐98 level while the miR‐98 inhibitors group decreased miR‐98 level (both P < 0.01). Dual luciferase reporter showed BMP2 was the target gene of miR‐98. The levels of mRNA and protein expression of BMP2, protein expression of RUNX2, alkaline phosphatase activity and osteocalcin content significantly decreased in the miR‐98 mimics group while increased in the miR‐98 inhibitors group and showed no changes in the NC group and Mock group (all P < 0.05). The miR‐98 mimics group showed obviously declined stained red particles and the miR‐98 inhibitors group showed opposite result. After lowering the expression of miR‐98, osteogenic differentiation ability of hBMSCs rose, which was weakened by the transfection with siBMP2. miR‐98 may regulate osteogenic differentiation of hBMSCs by targeting BMP2.

In-vitro characterization of canine multipotent stromal cells isolated from synovium, bone marrow, and adipose tissue: a donor-matched comparative study

“…In support of these potential explanations, pilot studies in our laboratory revealed that canine BMP-2 is transcribed at an extremely low level under control or traditional osteogenic conditions in early cMSC osteogenic cultures, requiring greater than 35 cycles to reach detection levels using qPCR (unpublished observations). In contrast, human MSCs robustly transcribe BMP-2 under similar conditions [ 150 – 153 ]. In-vivo evidence supporting the relevance of our in-vitro findings also exists.…”

Common and rare variants of WNT16, DKK1 and SOST and their relationship with bone mineral density

“…According to various functional predictors, rs17143305 may affect the binding of miRNAs hsa-miR-541 and hsa-miR-4263 and has been reported as an eQTL for genes RP11-3L10.1 and FAM3C in different tissues. Interestingly, hsa-miR-541 has been described as a negative regulator of osteoblastic differentiation 28 , in the only study in which it is studied in relation to bone tissues, to the best of our knowledge. On the other hand, as explained above, FAM3C was reported to have a negative effect on osteogenic differentiation 23 .…”

Small non-coding RNA landscape of extracellular vesicles from human stem cells

“…mir125b directly targeting BMPR1b was identified as negative regulator of osteogenic differentiation of human BM-MSC 63 . mir-98 in BM-MSCs regulated osteogenic differentiation by targeting BMP2 64 . mir-195 not only acts as a negative regulator of osteogenesis in human BM-MSCs, but also as an inhibitor of stem cell proliferative capacity and as an anti-angiogenic by targeting VEGF 65 .…”