Acne vulgaris is one of the most common dermatological diseases, and its pathogenesis is multifactorial. Although there are some reports about role of oxidative stress biomarkers in blood, serum, plasma in the pathogenesis of acne, there has been no report about oxidative stress biomarkers in scraping samples. For this reason, the aim of our study is to determine the role of oxidative stress biomarkers and adenosine deaminase (ADA) in scraping samples of acne lesion and to determine a possible link with the clinical severity. Fifty patients with different severity of acne vulgaris and forty healthy controls were enrolled. In both groups, ADA and oxidative stress biomarkers such as the levels of malondialdehyde (MDA), and reduced glutathione (GSH), the activities of superoxide dismutase (SOD) and catalase (CAT) in scraping samples were measured spectrophotometrically. The levels of SOD, CAT, GSH, MDA, and ADA of patients with acne were higher than the control patients (P < 0.05). ADA and MDA levels in patients with severe acne were significantly higher about two- to four-fold compared with other acne groups (P < 0.05). However, CAT, SOD, and GSH levels were lower in patients with severe acne vulgaris than the others (P < 0.05). Increased antioxidant enzyme levels of scraping samples in patients might be a local response of the organism to increased oxidative stress. Also, we believe that oxidative stress increases in parallel with the disease severity, hence antioxidant therapy may be beneficial if given in addition to the routine treatment of severe acne.