Background: The personal protective equipment (PPE) and frequent hand hygiene procedures needed during the COVID-19 outbreak impair skin integrity in healthcare workers (HCWs). We aimed to evaluate the prevalence and risk factors of adverse skin reactions related to infection-prevention measures among HCWs. Methods: A questionnaire survey was administered to evaluate the duration of PPE, disinfectant, and moisturizing agent use, as well as handwashing frequency among our hospital’s HCWs. Results: The questionnaire was completed by 702 HCWs with a mean age of 34.8±9.8 years. Adverse skin reactions were reported by 79.5% of our participants. Hands (63.5%) and face (48.9%) were the most commonly affected areas. Female sex, being a doctor/nurse, having a history of underlying chronic dermatoses, and PPE usage more than six hours per day were increased the risk of adverse skin reactions. Handwashing more than 10 times/day and moisturizing less than 5 times/day were also related to increased adverse skin reactions. In HCWs, wearing more than one mask was associated with pressure-induced skin changes on the face and triggering herpes labialis. Conclusion: Hand hygiene-associated dermatitis is triggered by frequent handwashing and less moisturizing among HCWs. Surgical masks may also be just as responsible as N95 masks for causing facial skin damage.