To evaluate extracellular matrix levels, 3 × 105 NP cells were suspended in 20 µl of complete medium (Gibco, Thermo Fisher Scientific, MA, USA) and seeded at the center of a 24-well plate. After 2 h of incubation at 37 °C, 0.5 ml of complete medium (Gibco, Thermo Fisher Scientific, MA, USA) was added. After an additional 24 h of incubation, the following treatments were performed. After 3 d, the cells were subjected to staining with toluidine blue solution (Solarbio, Beijing, China) and alcian blue solution (Solarbio, Beijing, China) for 1 h at room temperature. In this experiment, NP cells were respectively treated with IL-1β and IL-1β + mannose.
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