The increase in antibiotic resistance requires effective non-antibiotic therapies for acne. Cold atmospheric plasma (CAP) inactivates bacteria and improves wound healing, but its effect on acne has not been investigated. The objective of this controlled split-face study was to assess safety and efficacy of CAP in moderate acne. One side of the face received 8-10 treatments with cold helium plasma within 4-6 weeks; follow-up was two and four weeks thereafter. Acne lesions were counted, followed by global acne severity ratings. Of the 34 patients included, 29 completed the study. No serious adverse events occurred. The two facial sides did not significantly differ in the number of inflammatory and non-inflammatory lesions. An interaction effect of number and type of treatment was found for inflammatory lesions. Lesion reduction after 10 treatments was significantly higher on the treated than on the untreated side. Percentage of patients reporting improved aesthetics was higher for the treated than for the untreated side after treatment completion (79% vs. 45%) and at the two- (72% vs. 45%) and four-week follow-up (79% vs. 52%). In conclusion, CAP was safe with excellent tolerability, showed moderate reduction in acne lesions and led to higher patient-based ratings of aesthetics than non-treatment.