Objective To investigate the effect of exosomal miR-5106 from M2 macrophages under shear stress on differenti-ation of bone marrow mesenchymal stem cells (BMSCs) into osteoblasts.Methods Exosomes were isolated from M2 macrophages under shear stress and normal conditions.Transmission electron microscopy (TEM) was used to observe the morphological character-istics of exosomes,and Western blotting was performed to detect exosome-specific proteins TSG101,CD63,and Alix.Bioinformatics analysis was used to identify osteogenesis-related differential miRNAs and their binding sites to the BMP-2 promoter.After transfec-tion with differential miRNAs,BMP-2 expression levels were assessed by immunofluorescence staining,and qRT-PCR was performed to validate the expression changes of osteogenesis-related factors in BMSCs.Results miR-5106 was found to bind to the BMP-2 promoter.Immunofluorescence staining showed that the miR-5106 mimic enhanced BMP-2 immunoreactivity,while the miR-5106 inhibitor reduced BMP-2 immunoreactivity.qRT-PCR analysis revealed that the miR-5106 mimic upregulated the mRNA expression of osteogenesis-related factors ALP,BMP-2,OSX,and RUNX2,whereas the miR-5106 inhibitor suppressed their expression.Conclu-sion Shear stress regulates differentiation of BMSCs into osteoblasts via M2 macrophage-derived exosomal miR-5106.
维普
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