Journal: Inflammation

Article Title: Loss of the miR-221/222 Cluster Promotes the Pathogenesis of Osteoarthritis in Inflamed Mouse Chondrocytes and Osteoarthritis Models

doi: 10.1007/s10753-025-02411-4

Figure Lengend Snippet: Downregulation of miR-221 and miR-222 expression in OA tissue and IL-1β-stimulated chondrocytes. ( A-B ) Safranin O staining of the knee joints of OA model mice post-DMM surgery at the indicated time point showed progressive deterioration of articular cartilage. ( C ) Decreased expression of miR-221 and miR-222 in cartilage tissues from mouse joints ( n = 5 biologically independent samples). ( D ) Validation of in vitro OA model induction: NO levels were measured in the supernatant of IL-1β (10 ng/ml)-stimulated chondrocytes, along with mRNA expression levels of MMP-13 and COL2A1 in the same cells ( n = 5 biologically independent samples). ( E-F ) Time- and dose-dependent reduction in miR-221 and miR-222 expression in chondrocytes treated with IL-1β ( n = 5 independent biological samples). Data are presented as mean ± SEM. p values were calculated using one-way ANOVA followed by Tukey’s post hoc test

Article Snippet: After that, cells were incubated with anti-MMP-13 or COL2A1 antibodies (Both 1:500, Bioss), followed by FITC-conjugated goat anti-rabbit IgG (1:500, Proteintech).

Techniques: Expressing, Staining, Biomarker Discovery, In Vitro

Journal: Inflammation

Article Title: Loss of the miR-221/222 Cluster Promotes the Pathogenesis of Osteoarthritis in Inflamed Mouse Chondrocytes and Osteoarthritis Models

doi: 10.1007/s10753-025-02411-4

Figure Lengend Snippet: Exacerbation of OA progression in miR-221/222 cluster KO mice. ( A ) X-ray images of mouse joints 8 weeks post-DMM showing increased osteophytes, irregular joints, and pronounced joint stenosis in the KO + DMM group compared with the WT + DMM group. ( B-F ) Histological staining of joint sections at 8 weeks post-DMM confirms more severe degradation of cartilage and synovial fibrosis in the KO + DMM group ( n = 5 independent biological samples). ( G-K ) Western blot analysis indicates elevated type I collagen expression ( G ), while immunofluorescence analysis shows reduced COL2A1 levels ( H , I ) and increased MMP-13 expression ( J , K ) in KO + DMM joints at 8 weeks post-surgery ( n = 3 independent biological samples). For all the graphs, the values represent the mean ± SEM. p values were calculated via one-way ANOVA, followed by Tukey’s post hoc test

Article Snippet: After that, cells were incubated with anti-MMP-13 or COL2A1 antibodies (Both 1:500, Bioss), followed by FITC-conjugated goat anti-rabbit IgG (1:500, Proteintech).

Techniques: Staining, Western Blot, Expressing, Immunofluorescence

Journal: Inflammation

Article Title: Loss of the miR-221/222 Cluster Promotes the Pathogenesis of Osteoarthritis in Inflamed Mouse Chondrocytes and Osteoarthritis Models

doi: 10.1007/s10753-025-02411-4

Figure Lengend Snippet: Increased inflammatory mediator levels and the expression of chondrocyte OA-related factors in miR-221/222 cluster KO mice. ( A-C ) Levels of IL-6 ( A ), TNF-α ( B ), and NO ( C ) production in IL-1β (10 ng/ml) treated chondrocyte-free supernatant from WT and miR-221/222 cluster KO mice ( n = 5 independent biological samples). ( D-E ) Expression of inflammation-related proteins (COX2 and iNOS), OA-associated proteins (MMP-13), and COL2A1 in IL-1β (10 ng/ml) treated chondrocytes from WT and miR-221/222 cluster KO mice ( n = 3 independent biological samples). ( F-I ) Immunofluorescence confirming COL2A1 ( F and G ) and MMP-13 ( H and I ) expression in IL-1β (10 ng/ml) treated chondrocytes from WT and miR-221/222 cluster KO mice ( n = 5 independent biological samples). For all the graphs, the values represent the mean ± SEM. p values were calculated via one-way ANOVA, followed by Tukey’s post hoc test

Article Snippet: After that, cells were incubated with anti-MMP-13 or COL2A1 antibodies (Both 1:500, Bioss), followed by FITC-conjugated goat anti-rabbit IgG (1:500, Proteintech).

Techniques: Expressing, Immunofluorescence

Journal: Inflammation

Article Title: Loss of the miR-221/222 Cluster Promotes the Pathogenesis of Osteoarthritis in Inflamed Mouse Chondrocytes and Osteoarthritis Models

doi: 10.1007/s10753-025-02411-4

Figure Lengend Snippet: Alleviation of OA progression by the administration of agomir miR-221/222. ( A ) Schematic of the experimental design. ( B–D ) Intra-articular injection of the miR-221/222 agomir for 8 weeks attenuated OA progression in DMM-induced WT mice, as evidenced by improved histological staining ( B ) and decreased MMP-13 expression in joint tissues ( C , D ). E–F) Treatment also enhanced COL2A1 expression levels in the joints. ( n = 9 biologically independent samples). Data are presented as mean ± SEM. p values were determined using one-way ANOVA followed by Tukey’s post hoc test

Article Snippet: After that, cells were incubated with anti-MMP-13 or COL2A1 antibodies (Both 1:500, Bioss), followed by FITC-conjugated goat anti-rabbit IgG (1:500, Proteintech).

Techniques: Injection, Staining, Expressing