fig7
Figure 7. miR-21a-5p alleviated DMM-induced OA progression in vivo. (A) Schematic timeline of the in vivo study designed to evaluate the role of miR-21a-5p in macrophage efferocytosis, inflammatory responses, and cartilage injury; (B and C) Representative micro-CT reconstructions of mouse knee joints and corresponding assessment of osteophyte formation. Osteophytes are highlighted in yellow (n = 5 mice per group); (D) Representative SO/FG-stained sections of knee joints from mice receiving the indicated treatments with or without miR-21a-5p intervention. Scale bars: 1,500 and 20 μm; (E) Quantitative evaluation of OARSI, synovitis, and osteophyte scores based on histological staining (n = 5 mice per group); (F) Representative immunofluorescence images showing iNOS expression in mouse knee synovial tissue. Scale bar: 20 μm; (G) Quantitative assessment of iNOS immunofluorescence signals (n = 5 mice per group); (H) Representative TUNEL-stained images of mouse articular cartilage. Scale bar: 20 μm; (I) Quantification of TUNEL-positive cells in cartilage (n = 5 mice per group); (J) Representative immunofluorescence images demonstrating Aggrecan and Mmp3 expression in mouse articular cartilage. Scale bar: 20 μm; (K and L) Quantitative measurements of Aggrecan and Mmp3 immunofluorescence signals, respectively (n = 5 mice per group); (M) Representative immunofluorescence images illustrating YAP1 expression in mouse knee synovium. Scale bar: 20 μm; (N) Quantitative analysis of YAP1 immunofluorescence intensity (n = 5 mice per group); (O) Yap1 expression in mouse knee synovial tissue was determined by RT-qPCR (n = 3 mice per group). Osteophyte maturity, OARSI, synovitis, and osteophyte scores were analyzed using the Mann-Whitney U test. Continuous outcomes involving multiple experimental groups were evaluated by two-way ANOVA followed by Šidák’s multiple-comparison test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. DMM: Destabilization of the medial meniscus; OA: osteoarthritis; micro-CT: micro-computed tomography; SO/FG: Safranin O/Fast Green; OARSI: Osteoarthritis Research Society International; iNOS: inducible nitric oxide synthase; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; RT-qPCR: quantitative real-time polymerase chain reaction; NC: negative control; ANOVA: analysis of variance.
