Adamts 4

Sections were deparaffinized and hydrated and then placed in a citric acid solution to undergo 16 hours of incubation in a 60°C water bath for antigen retrieval, followed by incubation in a hydrogen peroxide solution for 10 minutes and blocking in 10% goat serum at 37°C for 30 minutes. The sections were then incubated in antibodies against p65, ADAMTS‐4 (purchased from ABclonal, Wuhan, China), Col1a2, MMP‐13, HIF‐1α (purchased from Abcam, Cambridge, MA, USA), and Col2a1 (purchased from Millipore, Burlington, MA, USA) at 4°C overnight, followed by incubation in secondary antibody (purchased from ABclonal, Wuhan, China) for 1 hour at room temperature, DAB chromogenesis, hematoxylin staining, and mounting to observe the expression of the corresponding proteins under a microscope.

MT, Rat IL-1β, 4′,6-diamidino-2-phenylindole (DAPI), collagenase type II, and EX527 were purchased from Sigma Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) (BI, Kibbutz Bet Hamek, Israel), Dulbecco’s Modified Eagle’s Medium-(DMEM-) F12 Medium, and Phosphate Buffered Saline purchased from Corning (New York, NY, USA). 1% penicillin/streptomycin, cell counting kit-8 (CCK-8), and staining kit were purchased from Beyotime (Haimen, China). Trypsin (0.25%) was purchased from Gibco (Grand Island, NY, USA). Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China). Horseradish peroxidase-conjugated goat anti-mouse/rabbit immunoglobulin G (IgG) secondary antibody was purchased from ABclonal Technology (UK). GAPDH was purchased from CST (Danvers, MA, USA).