Objective To investigate the changes in microRNA expression profiles within apoptotic vesicles derived from BMSCs under simulated negative pressure environments, and to provide a theoretical basis for the mechanism by which mechanical stress microenvironments influence the progression of osteoarthritis. Methods A negative pressure cellular environment was established using a pressure-loading system. Cell viability and apoptosis were assessed via CCK-8 assay, Western blotting, and Annexin V-FITC/PI double staining. Apoptotic vesicles were isolated by differential centrifugation and characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. Using the Hiseq Single End mode for small RNA sequencing analysis, differential expression analysis of microRNAs was performed using DESeq to screen for differentially expressed microRNAs.Real time quantitative PCR validation of differentially screened microRNAs.After treating BMSCs with differential microRNA inhibitors, the effects of the screened differential microRNAs on BMSCs were detected. Results Compared to apoptotic vesicles generated by bone marrow mesenchymal stem cells (BMSCs) under STS chemical treatment, those produced under a -40 kPa pressure environment exhibited significantly upregulated miR-183-5p and downregulated miR-3473. GO and KEGG enrichment analysis revealed that differential microRNAs affect cell activity and inflammatory response through multiple signaling pathways.Inhibition of miR-183-5p and miR-3473 expression reduces the proliferation activity of BMSCs. After inhibiting the expression of miR-183-5p, the expression levels of inflammatory factors increased. Inhibition of miR-3473 expression, unchanged IL-6 expression level, and significant increase in TNFα expression level. Conclusions microRNAs specifically expressed in apoptotic vesicles derived from bone marrow mesenchymal stem cells (BMSCs) under negative pressure stimulation may act as critical mechanical signaling mediators, modulating inflammatory response processes to participate in the pathogenesis and progression of arthritis.