Ozonized autohemotransfusion has been used as a complementary therapy in patients with peripheral arterial disease (PAD). To determine whether ozone therapy could acutely modify artery vasodilatory capacity, a flow-mediated dilation test was performed at the brachial artery level before and after an ozonized autohemotransfusion in 16 patients with PAD, mean (± SD) age 55±1.8 years, and 14 healthy volunteers matched for age, sex and body mass index. Before ozonized autohemotransfusion, the mean baseline diameter of the brachial artery was higher in PAD patients than in healthy subjects (4.6±0.54 mm versus 3.6±0.54 mm, P<0.001) while mean flow-mediated brachial artery dilation and percentage of increase in flow were significantly lower in PAD patients than in controls (6.3±6.1% versus 11.8±2.4%, P<0.02; 433±61% versus 580±46%, P<0.02, respectively). No significant changes were observed after ozonized autohemotransfusion, indicating that ozonized autohemotransfusion does not modify endothelium-dependent ischemia-induced vascular reactivity.