The in vivo genotoxicity of isotretinoin assessed by comet assay
Keywords:COMET assay, Isotretinoin
Isotretinoin (13-cis-retinoic acid) is a vitamin A derivative, which is used as the most effective medication in the treatment of skin acne. The evidence related to the genotoxicity of isotretinoin are controversial. While inducing apoptosis and necrosis at higher doses in in vitro studies, isotretinoin had no genotoxic effect when tested in human lymphocytes. It has been reported that patients with cystic acne showed elevated level of plasma biomarker of DNA oxidation, after treatment by isotretinoin. However, the controversial effect of isotretinoin on frequency of micronuclei was reported. Thus, further elucidation of isotretinoin biological effects may help to reach a conclusion on its use. Therefore, the present study was carried out. The comet assay is a technique that can measure DNA damage such as single strand breaks, double strand breaks, crosslinks and base damage for individual eukaryotic cells. The purpose of this study is to evaluate the extent of DNA damage in leukocytes of patients on isotretinoin treatment. To this aim, a fresh blood sample was obtained from 23 volunteers, including 12 healthy individuals and 12 patients with acne on isotretinoin treatment. After preparation of the slides, images of 100 randomly cells were captured per sample (Nikon fluorescence microscope) and DNA damage was evaluated by measuring the tail length (TL), tail DNA percent (TD), and tail moment (TM) using TriTek CometScore V 2.0 software. Statistical analysis indicated that no significant difference in TL, TD and TM parameters was found between two groups. The results of the present study did not show the genotoxicity effect of isotretinoin on the patient’s leucocytes.
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