Il 12

After treatment of the chondrocytes with KBH-JP-040 (0, 50, 100, and 200 μg/mL) for 1 h and following co-incubation for 24 h with rhIL-1α (5 ng/mL) and aggrecan (MyBioSource, San Diego, CA, USA), collagen type-II content was assessed using a sandwich enzyme-linked immunosorbent assay (ELISA) kit (Chondrex, Inc., Redmond, WA). Prostaglandin E2 (PGE2) (R&D Systems Inc., Minneapolis, MN, USA). The levels of MMP-1, MMP-9 (R&D Systems Inc.), MMP-13 (MyBioSource), IL-6, IL-10 (eBioscience, San Diego, CA, USA), and IL-12 (MyBioSource) protein in culture medium were assayed with an ELISA kit (R&D Systems Inc.) following the manufacturer’s instructions.

Cytokines in the BAL and serum were quantified with enzyme-linked immunosorbent assays (ELISA) following the manufacturer’s instructions of the following kits for the guinea pigs: TNF-α (MBS9303082; MyBioSource, San Diego, CA, USA), IFN-γ (MBS701377; MyBioSource, San Diego, CA, USA), TGF-β1 (CSB-E06773GU; CUSABIO TECHNOLOGY LLC; Houston, TX, USA), IL-1β (MBS765173; MyBioSource, San Diego, CA, USA), 1L-6 (MBS269054; MyBioSource, San Diego, CA, USA) IL-8 (MBS282965; MyBioSource, San Diego, CA, USA), IL-12 (MBS704591; MyBioSource, San Diego, CA, USA).