miR-3960 from Mesenchymal Stem Cell-Derived Extracellular Vesicles Inactivates SDC1/Wnt/β-Catenin Axis to Relieve Chondrocyte Injury in Osteoarthritis by Targeting PHLDA2

Figure 2

MSCs-EVs carrying miR-3960 inhibits chondrocyte injury in OA. (a) Venn diagram of 134 downregulated miRNAs (red circle) in GSE105027 dataset and the top 30 miRNAs enriched in MSCs-EVs (blue circle) and miR-3960 expression in GSE105027 dataset. (b) miR-3960 expression in the cartilage tissues of OA patients () and patients with traumatic amputation () determined by RT-qPCR. (c) miR-3960 expression in the IL-1β-treated chondrocytes determined by RT-qPCR. (d) The expression of miR-3960 in MSCs-EVs was determined by qRT-PCR ( vs. that in MSCs). (e) Transfer of Cy3-labeled miR-3960 into chondrocytes via MSCs-EVs (). (f) miR-3960 expression in the chondrocytes treated with IL-1β+MSCs-EVs determined by RT-qPCR. (g) miR-3960 expression in the chondrocytes treated with miR-3960 mimic determined by RT-qPCR. (h) miR-3960 expression in the chondrocytes treated with IL-1β+MSCs-EVs-miR-3960 mimic determined by RT-qPCR. Chondrocytes were treated with IL-1β and/or MSCs-EVs-miR-3960 mimic. (i) Cell proliferation assessed by CCK8 assay. (j) Apoptosis assessed by flow cytometry. (k) Protein levels of collagen II, aggrecan, MMP13, and ADAMTS5 in chondrocytes measured by immunoblotting. (l) Levels of IL-6 and TNF-α in chondrocytes measured by ELISA. vs. patients with traumatic amputation, chondrocytes without treatment, chondrocytes treated with mimic-NC, or chondrocytes treated with MSCs-EVs-mimic-NC; ^# vs. chondrocytes treated with IL-1β; & vs. chondrocytes treated with IL-1β+MSCs-EVs-mimic-NC. Data are shown as the of three technical replicates. Data between two groups were compared by independent -test. Data comparisons among multiple groups were analyzed by the one-way ANOVA with Tukey’s post hoc test, and comparison of data at different time points was analyzed using repeated measurement ANOVA with Tukey’s post hoc test.

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