Cytotoxicities and Anti-Fibrotic Effects of Pirfenidone and Mitomycin C on Human Fibroblasts

Purpose: The cytotoxicities and anti-fibrotic effects of mitomycin C and pirfenidone on human dermal fibroblast were evaluated. Methods: Initially, 24-hour cell cultures were exposed to transforming growth factor (TGF)-β1, different concentrations of mitomycin C, and pirfenidone solutions in order to evaluate cytotoxicity. Expressions of fibronectin, collagen type 1, α smooth muscle, and β-actin were evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR) and western blot in mitomycin C solutions at concentrations of 4 μg/mL and 20 μg/mL, and in pirfenidone solutions at 250 μg/mL and 500 μg/mL. Results: In comparison to cell cultures exposed to TGF-β1 solutions, cytotoxicities were increased in solutions of mitomycin C at 4 μg/mL, 20 μg/mL, 40 μg/mL and pirfenidone at 500 μg/mL, 750 μg/mL, 1,000 μg/mL (p < 0.05, Mann Whitney U-test). The results of real-time RT-PCR show that expressions of fibronectin, collagen type 1, and α smooth muscle were significantly more decreased in all concentrations of mitomycin C and pirfenidone compared to those in TGF-β1 solution. In western blot analysis, expressions of fibronectin and α smooth muscle were decreased in all concentrations of mitomycin C and pirfenidone compared to TGF-β1 solution. Conclusions: Both drugs have cytotoxicities and anti-fibrotic effects, but pirfenidone was found to have less cytotoxicity and mitomycin C was found to have more anti-fibrotic effects when compared to each other. J Korean Ophthalmol Soc 2014;55(7):1077-1083