The comet assay a method to measure DNA damage in oral submucous fibrosis patients: A case-control study

Abstract

Context: Oral submucous fibrosis (OSF) is a chronic, progressive condition and rise in incidence of this disease has been attributed to an increased uptake of betel nut and its products, especially among young people. Due to its potential for malignant transformation, easy and reliable techniques for its early detection are needed. Comet assay is a simple technique to detect and quantify single strand breaks, double strand breaks and alkali labile damages. Aims: The present pilot study was undertaken to assess the deoxyribonucleic acid (DNA) damage in OSF and healthy groups using comet assay. Settings and Design: The hospital-based study was conducted by evaluating buccal mucosal cells of 50 individuals in the age range of 16-47 years and comet assay was carried out for all the two groups under alkali conditions. Materials and Methods: Oral epithelial cells were obtained from buccal mucosa of each 25 cases of healthy and OSF groups were subjected to comet assay. DNA damage was evaluated by measuring the tail length. Statistical Analysis Used: Student's t-test and Turkeys-multiple post-hoc procedures were used to statistically analyze the obtained data. Results: Increase in the tail length in buccal epithelial cells of OSF group when compared with the healthy group was noted. Furthermore, there was a significant increase in the DNA damage with duration of habits. Conclusions: We conclude that increase in the tail length formation in OSF groups when compared to healthy groups; this indicates DNA damage in the oral epithelial cells. The comet assay is a relatively simple, but sensitive and well-validated tool for measuring strand breaks in DNA in single cells.