Interleukin 1β il 1β

All chemicals and reagents were obtained commercially and were analytical grade. Hexadecyl trimethyl ammonium bromide (CTAB), tetraethyl orthosilicate (TEOS), 3-glycidoxypropyltrimethoxysilane (GPTMS), aqueous ammonia, sodium carbonate, ethanol, acetic acid and methyl alcohol were obtained from Sinopharm Chemical Reagent Co, China.
0.2% trypsin was obtained from Hyclone, USA. Dulbecco’s modified Eagle’s medium (DMEM) and 1% penicillin/streptomycin were obtained from Thermo Fisher Scientific, USA. Monosodium iodoacetate (MIA) was obtained from Aladdin Industrial Co, China. 10% fetal bovine serum (FBS) and 5% bovine serum albumin (BSA) were obtained from Jackson Immuno Research Inc. USA. Phosphorylated p65, p65, phosphorylated IKKβ, IKKβ, phosphorylated IκBα, IκBα primary antibodies were obtained from Cell Signaling Technology, USA. Collagenase, Celastrol (CSL) and interleukin 1β (IL-1β) were obtained from Sigma-Aldrich, USA. Ethylenediaminetetraacetic acid (EDTA), 10% paraformaldehyde, horseradish peroxidase (HRP)-conjugated secondary antibody, Cell Counting Kit-8 (CCK-8), RIPA lysis buffer, BCA Protein Assay Kit and enhanced chemiluminescence (ECL) detection kit were obtained from Beyotime Institute of Biotechnology, China.

The coding sequence of PART‐1 and TGFBR2 was subcloned into the pcDNA3.1 (+) vector (Invitrogen). PART‐1 siRNA, TGFBR2 siRNA, nonsense siRNA, miR‐590‐3p mimic, miR‐590‐3p inhibitor and respective controls were purchased from Guangzhou Ribobio company. Chondrocytes were transfected with the above plasmids, siRNAs or miRNAs using Lipofectamine 3000 (Invitrogen), and transfected chondrocytes were collected for in vitro assays at 24‐72 hours after transfection. In some experiments, the cells were stimulated with 5 ng/mL interleukin 1β (IL‐1β; Sigma‐Aldrich) for 24 hours.