Extracellular vesicles derived from synovial inflammatory macrophages induce the onset of osteoarthritis in a FMRP-selectively sorted manner [RNA-seq]

Purpose:Next Generation Sequencing has revolutionized the effect of EVs secreted by synovial inflammatory macrophages on the cartilage of OA progression. The goals of this study are to compare NGS-derived cartilages transcriptome profiling (RNA-seq) between exosomes derived from synovial inflammatory macrophages and PBS treated OA rats model. The key biological processes and siginaling pathway were identified with immunofluorescence (IF), Transmission electron microscopy (TEM) and to evaluate protocols for optimal high-throughput data analysis. Overall design: Methods: 8-week old SD rats undergoing DMM+ACLT operations allocated into two distinct categories: OA + PBS group; OA + PBS-EVsM1 group. Three weeks after the surgery, rats received several injections of 50 µl PBS or 50 µl PBS-EVsM1 (1×1010 particles/mL) intra-articularly throughout the next 6 weeks (twice a week). After 10 weeks of surgery, the rats' anesthesia was kept up with isoflurane, and the specimens were collected. Cartilages mRNA profiles of OA rats treated with PBS or EVs derived from synovial inflammatory macrophages (EVsM1) were generated by deep sequencing, in replicate, using illumina NovaseqTM 6000 platform. The bioinformatics examination, comprising genes differentially expressed (DEGs), KEGG and GSEA enrichment. Immunofluorescence (IF), Transmission electron microscopy (TEM) were performed to evaluate the optimal high-throughput data analysis.