Abstract: The ozonides produced by ozonation procedure have significant killing effects on bacteria, fungi and yeasts. They have effects on activation of local microcirculation, tissue growth, and on other processes involved in the treatment of skin disorders. The purpose of present study is to formulate and evaluate the healing effects of ozonated extra virgin olive oil on acne vulgaris as a single-blind, non randomized clinical trial. Each patient received a placebo on his/her lesions for one month and after 10 days of washouts, they were treated with ozonated olive oil applied on their whole lesions for one month. The results were expressed as the percentages of reduced inflammatory versus non-inflammatory lesions. After one month of ozonated olive oil treatment, a seventy-five percent healing was observed in 50% of patients while under the same condition the remaining twenty five percent of the patients had 50% healing. After one month of applying ozonated olive oil, both inflammatory and noninflammatory acne lesions were decreased by 65%. In conclusion, topical application of ozonated olive oil has shown significant healing effects on patients suffering from acne vulgaris. Biocompatibility behavior of ozonated olive oil makes it an excellent candidate to be prescribed as a highly potential anti-acne medicine