OBJECTIVE The objective of this study was to investigate the association between 11 patient safety culture dimensions and the implementation of 7 organizational patient safety defenses. DESIGN Data were gathered within a cross-sectional, retrospective survey. SETTING Emergency departments (EDs) in the Netherlands. PARTICIPANTS Thirty-three EDs of non-academic hospitals, which belong to the clientele of Dutch largest medical liability insurer. MAIN OUTCOME MEASURES Implementation of the separate organizational patient safety defenses (0 = insufficient/sufficient, 1 = good). RESULTS Analyses showed that several culture dimensions were negatively or positively associated with the implementation of the patient safety defenses. A culture in which hospital handoffs and transitions were perceived adequate was related to less frequent implementation of four of seven organizational patient safety defenses, whereas a culture with well-perceived hospital management support for patient safety predicted more frequent implementation of four of seven organizational patient safety defences. CONCLUSIONS Results suggest that well-perceived culture dimensions might inhibit improvements by lack of a sense of urgency as well as facilitate improvements by inducing feelings of support for organizational changes and improvements. The influence of patient safety culture appeared to be not always as straightforward as it seems to be in advance.