MiR-181d-5p regulates implant surface roughness-induced osteogenic differentiation of bone marrow stem cells

Materials Science and Engineering: C(2021)

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摘要
Constructing moderate surface roughness is a widely used, non-toxic, cost-effective, and outcome-predictable approach to accelerate implant osteointegration in clinical settings. MicroRNAs (miRNAs) play vital regulatory roles in the osteogenic differentiation of bone marrow stem cells (BMSCs). However, their specific contribution to the influence of surface roughness on osteoblastic behavior remains unknown. Therefore, applying the smooth titanium surface as a control, a typical titanium surface with moderate roughness was prepared here to reveal the mechanism through which surface roughness regulates cell osteogenic behavior by altering miRNA expression. First, the morphology and roughness of two surfaces were characterized, and the enhanced osteogenic differentiation of BMSCs on rough surfaces was verified. Then, twenty-nine differentially expressed miRNAs in BMSCs cultured on different surfaces were selected via miRNA chip and corresponding functional prediction. After verifying the expression of these miRNAs using quantitative real-time polymerase chain reaction, four were considered eligible candidates. Among these, only miR-181d-5p significantly affected RUNX2 gene expression based on overexpression and knockdown experiments. From the osteogenesis-related gene and protein expression, as well as alkaline phosphatase and alizarin red experiments, we further confirmed that the downregulation of miR-181d-5p promoted osteogenic differentiation of BMSCs, and vice versa. In addition, rescue assays showed that the knockdown of miR-181d-5p improved the inferior osteogenesis observed on smooth surfaces, whereas the overexpression of miR-181d-5p suppressed the superior osteogenesis observed on rough surfaces. These results indicate that the moderate surface roughness of the implant stimulates the osteogenic differentiation of BMSCs by remarkably downregulating miR-181d-5p. These findings provide helpful information and a theoretical basis for the development of advanced implant materials for fast osteointegration.
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关键词
MicroRNAs,Surface roughness,Bone marrow stem cells,Osteogenic differentiation
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