Scrambled mirna agomir

An equal volume of mir-145-5p agomir or scrambled miRNA agomir (RiboBio, Guangdong, China) was injected in the tail vein of the mice on Day 30. Injected mir-145-5p agomir (n=4) or scrambled miRNA agomir (n=4) in the tail vein of mice at 1-week intervals. Four weeks after injection, the hind feet were used for micro-computed tomography (CT) analysis, and the ankles were used for histopathological analysis.

Male DBA/1 mice (15–20 g; 8 weeks old) were used in the present study. A total of 30 animals were housed at 24±1°C with a 12:12-h light/dark cycle and a relative humidity of 56±5%. Animals were provided access to food and water ad libitum, and were fed a commercial diet according to the guidelines of the Animal Ethical and Welfare Committee. All mice were housed individually. Lyophilized bovine type II collagen (Chondrex, Inc., Redmond, WA, USA) was dissolved overnight at 4°C in 0.05 M acetic acid under constant stirring. Next, the collagen was emulsified using Freund's complete adjuvant (Chondrex, Inc.) to provide a final concentration of 2 mg/ml. Mice were injected in the tail with 0.1 ml of emulsion on day 1 and booted intraperitoneally on day 21 with collagen. On day 30, an equal volume of mir-145-5p agomir or scrambled miRNA agomir (15 nM; Guangzhou RiboBio Co., Ltd.) was injected in the tail vein of mice at 1-week intervals. Four weeks after the first injection, the mice were sacrificed by injecting pentobarbital sodium at a dose of 100 mg/kg, then various indications such as respiration, heartbeat, pupil reflection, and nerve reflex were observed in the mice, and when all the aforementioned reactions disappeared, death was judged. Next, the right hind limb was dissected and synovial tissue was collected for subsequent experiments.