Fig. 6

Intraarticular administration of LPCAT3-siRNA using lipid nanoparticles ameliorates OA in mice. A OA was induced in knee tissues of mice by performing DMM surgery in 10-12 week-old C57WT mice, followed by intraarticular injection of LPCAT3 siRNA lipid nanoparticles (LPC3 siRNA) or scrambled siRNA (siRNA) or PBS as untreated control (sham) (n = 12 mice per group), administered immediately and thereafter at 1, 2, 4, and 6 weeks post-DMM surgery (5 injections total) at a dosage of 0.5 mg/kg body weight in 20 μl volume using an insulin syringe. B The synovial articular cartilage tissues were collected at 8 weeks post-DMM and the efficiency of knockdown of LPCAT3 was analyzed by immunoblotting. Blots represent data from 4 animals/groups. C The knee tissues were collected at 8 weeks post DMM and stained with Safranin O fast green (Scale bar- 50 µM). D, E The sections were graded in a blinded fashion by two independent observers and OARSI score (D) and synovitis score (E) were presented. F-J The frozen synovial cartilage tissues were homogenized in the RIPA buffer containing protease inhibitors and indomethacin (1 mg/mL), and centrifuged at 10,000g at 4°C to remove tissue debris. The supernatants were collected. The eicosanoids such as PGE₂ (F) and LTB₄ (G) and the cytokines such as TNFα (H), IL-6 (I), and MCP-1 (J) were analyzed by ELISA. Data represent mean ± SD of 2 independent experiments, a total of 8 animals in a group. ***P < .001.*P < .05