Figure 2.

Impact of nicotine on ihNPs cellular activity and chondrogenic markers. Initially, immortalized human nucleus pulposus cells (ihNPs) of IVD were treated with 100 µM, 500 µM and 1000 µM of nicotine for 7 days to evaluate its effects on proliferation ability (A). Owing to the increased deteriorating effect, concentration of 1000 µM nicotine was applied for the further experiments. (B) Gene and (C) protein expressions of chondrogenic markers including Sox9, Col II, and aggrecan. We further relatively quantified the expression of alcian blue in control and nicotine-treated ihNPs (D). The results are presented as mean ± S.D. (n = 3; *P < 0.05 and ***P < 0.001, respectively compared to control group).