Figure 1.

IL-1β treatment stimulated ROS production and mitochondrial dysfunction in chondrocytes. (A) and (B) Chondrocytes (1×10⁶ cells/well) were seeded in 6 well plate and treated with MitoSox Red (5μM) for 10 minutes followed by IL-1β for 5 minutes. Chondrocytes were analyzed for mitochondrial ROS by flow cytometer. (C) and (D) Chondrocytes were seeded in 6 well plates as above and treated with IL-1β for 5 minutes followed by incubation with JC-1 dye for 30 minutes. The green (FL1) and red (FL2) fluorescence of JC-1 as marker of ΔΨM loss was analyzed by flow cytometer (BD Accuri C6). CCCP was used as positive control. (E) Chondrocytes were either treated with IL-1β (or left untreated as control) and incubated at 37°C for 48 hours. Chondrocytes were stained with propidium iodide in 01% Triton X-100 and analyzed for pre-G1 population indicating cell death by flow cytometer. (F) Chondrocytes were treated as above and analyzed for cell death by TUNEL staining and analysis by flow cytometer. Data shown are the representative of three independent experiments from different patient’s samples, each done in triplicate. Values are expressed as mean ± SD (*p<0.05).