Cholesterol oxides are cytotoxic and have been implicated in many disease processes; however, it has been proposed that cholesterol oxides result from cholesterol acting as a sacrificial antioxidant. In this study, the effect of dietary cholesterol on DNA damage, assessed by the alkaline comet assay, was examined in male and female Syrian hamsters. Animals were fed ad libitum a modified AIN-76 diet (control) or a diet with 0.5% cholesterol for 10 weeks. Following the 10-week feeding period, there was no significant difference in body weight between cholesterol-fed and control animals. Cholesterol feeding resulted in significant liver hypertrophy, and increased plasma total and HDL cholesterol in both male and female animals compared with controls. There was no difference in liver cell DNA damage levels as measured by the comet assay. Heart cells from cholesterol-fed hamsters, however, showed a significant decrease in tail DNA (p = 0.050) indicating decreased damage compared with controls and a possible protective effect of cholesterol against DNA damage.