Fig 1: Therapeutic effects of sPLA2i-NPs for attenuation of DMM-induced chronic OA.(A) The study design of sPLA2i-NP treatment for DMM-induced OA mice. (B) Safranin O/Fast Green staining of sham- or DMM-operated knee joints with 2 or 4 months of treatment. Scale bar, 200 µm. (C) The OA severity was accessed by Mankin score at 2 months after surgery (n = 8). (D) Hematoxylin and eosin (H&E) staining of synovium tissue at 2 months after surgery. Scale bar, 100 µm. Red boxed areas indicate the synovial lining layer. (E) Synovial inflammation was evaluated (n = 8). (F) 3D color maps of femoral SBP thickness at 4 months after surgery. Color ranges from 0 µm (blue) to 320 µm (red). (G) Quantification of SBP thickness (Th.) at the medial posterior site (n = 8). (H) Osteophytes (red arrows) were revealed by 2D (top) and 3D (bottom) micro–computed tomography (micro-CT) images. (I) Osteophyte volume was quantified (n = 8). (J) von Frey assay was performed at 1, 2, 4, 8, and 12 weeks after surgery (n = 8). The data of day 0 were acquired before surgery. PWT, paw withdrawal threshold. (K) IHC staining of the tibial articular cartilage at 2 months after surgery. Scale bar, 100 µm. (L) Quantification of sPLA2-IIa–, p-P65–, and p-P100–positive chondrocytes (n = 5). Statistical analysis was performed using one-way ANOVA with Turkey’s post hoc test. Data presented as means ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001 in (C), (E), (G), (I), and (L). *P < 0.05 and **P < 0.01 for sPLA2i-NPs versus PBS in (J).
Fig 2: Therapeutic effects of sPLA2i-NPs for attenuation of load-induced PTOA.(A) The study design of sPLA2i-NP treatment for loading-induced OA mice. (B) Safranin O/Fast Green staining of the articular cartilage in sham or loaded (6 or 9 N) knee joints with 14-day treatment. Scale bar, 100 µm. Dashed lines indicate the range of loss of staining. (C) The length of cartilage lesion in sham and 6-N loaded knee joints was quantified. (D) TUNEL staining in the tibial articular cartilage from sham and 6-N loaded knee joints. Scale bar, 100 µm. (E) Quantification of TUNEL-positive chondrocytes (n = 5). (F) IHC staining of the tibial articular cartilage from sham and 6-N loaded knee joints. Scale bar, 100 µm. (G) Quantification of sPLA2-IIa–, p-P65–, and p-P100–positive chondrocytes (n = 5). (H) H&E staining of synovium tissue. Scale bar, 100 µm. Red boxed areas indicate the synovial lining layer. (I) Synovial inflammation was evaluated (n = 8). (J) von Frey assay at 1 and 2 weeks after 6-N loading (n = 8). The data of day 0 were acquired before loading. Statistical analysis was performed using one-way ANOVA with Turkey’s post hoc test. Data presented as means ± SEM. *P < 0.05 and ***P < 0.001 in (C), (E), (G), and (I) and *P < 0.05 for sPLA2i-NPs versus PBS in (J).
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