Study on the Applicability of Mytomycin C as a Positive Control Material for Agar Diffusion Cytotoxicity Test

Objective To find a non-volatile positive material control which showed moderately or severely cytotoxic and could be obtained easily. Methods The in vitro cytotoxicity study was conducted according to the agar diffusion method in ISO 10993.5-2009 and YY/T 0127.9-2001. Each sample was placed in contacted with the surface of an agar layer overlaying a monolayer of L-929 Mouse Fibroblast cells which were stained with neutral red vital stain at the bottom of the plate. As the sample, Mytomycin C was diluted to 2 mg/mL, 1 mg/mL and 0.5 mg/mL. Results In the group of 2 mg/mL Mytomycin C, it was observed that the cytotoxicity grade was 3. In the group of 1 mg/mL Mytomycin C, it was observed that the cytotoxicity grade was 3. In the group of 0.5 mg/mL Mytomycin C, it was observed that the cytotoxicity grade was 2. Conclusion 2 mg/mL Mytomycin C could be used as a positive material control of agar diffusion.