Patient safety culture is relevant both in the delivery of care and in the training of nursing staff, its purpose being to prevent and reduce risks associated with health care. This research aims to evaluate patient safety culture from the perspective of the nursing teams in a highly complex public hospital in the city of Valparaíso, Chile. A cross-sectional study with a quantitative approach applying descriptive, bivariate, and inferential statistical analysis was conducted on 259 nurses and nursing assistants from 13 adult medical–surgical units of the Carlos Van Buren hospital. The participants were obtained through a non-probabilistic convenience sample, answering the hospital survey on Patient Safety Culture version 2.0 (HSOPS 2.0), adapted to the Chilean population. The best-evaluated dimension was communication and receptivity; the worst was the support administrators provide for patient safety. This study identified the weaknesses and strengths of the hospital, the most worrying weakness being the shortage of human capital, material, and financial resources necessary to improve patient safety. This study was not registered.