In the realm of occupational accidents, the healthcare sector holds a significant position as it is classified as a highly hazardous workplace, thus bearing a heightened probability of accidents. Consequently, it becomes imperative to assess the occupational Health Literacy (OHL) of healthcare workers and identify the influencing factors. The aim is to determine the OHL levels of healthcare workers and examine their relationship with demographic, socio-cultural characteristics, and variables related to the work environment. This cross-sectional survey was conducted among 200 healthcare workers employed in healthcare services, administrative services, and support services at an education and research hospital in Niğde, Turkey, from November-December 2023. Data collection involved the utilization of a personal information form encompassing participants' socio-demographic characteristics and occupational health practices, along with the OHL Scale. Descriptive statistics, intergroup difference analyses, and multiple regression analysis were employed for data analysis. The participants demonstrated an average OHL score of 94.07 ± 11.33. They scored highest in Understanding occupational health and safety (OHS) Information (40.93) and lowest in Evaluating OHS Information (9.57). OHL among healthcare workers varied based on demographics, socio-cultural factors, and work-related variables. Age, education, tenure, vaccination, regular health screenings, workplace accidents and personal protective equipment (PPE) usage positively impacted OHL. Findings revealed participants possess a certain level of OHL. Age, education, tenure, vaccination, regular health screenings, workplace accidents and PPE usage accounted for 53% of OHL variance. Enhancing workplace safety measures and improving healthcare workers' OHL are recommended.