Background <p>Patient safety culture encompasses the values, beliefs, norms, and behaviours that influence patient safety. It is positively associated with improved patient outcomes and inversely related to adverse events. This study explores the patient safety culture in the operating theatre setting to support national efforts aimed at mitigating adverse events. Specifically, it assesses the patient safety culture in six Norwegian hospitals and potential associations between scores from the Norwegian Hospital Survey on Patient Safety Culture 2.0 (N-HSOPSC 2.0) scores and socio-demographic factors, including gender, field of expertise, years of experience, leadership responsibilities, and hospital trust.</p> Methods <p>An exploratory cross-sectional multicentre study using the N-HSOPSC 2.0 was conducted between 2021 and 2022 among operating theatre staff across six Norwegian hospitals in three hospital trusts. Descriptive statistics were used to report the scores of 10 factors and three outcome variables. Generalised linear models were employed to analyse the relationships between socio-demographic variables (gender, field of expertise, years of experience, leadership responsibilities and hospital trust) and the N-HSOPSC 2.0 scores.</p> Results <p>A total of 217 individuals responded to the questionnaire. The teamwork factor had the highest percentage of positive responses (85.8%), while all other factors showed less than 75% positive responses, indicating an unsatisfactory patient safety culture. Notably, three factors had less than 50% positive responses, underscoring the need for targeted improvement initiatives. Respondents with leadership responsibilities reported significantly more positive perceptions of the patient safety culture on three factors and one outcome variable compared to those without such responsibilities. Similarly, individuals identified as unit leaders or administrators reported significantly more positive perceptions of four patient safety culture factors and a more favourable overall patient safety rating. Perceptions varied across different fields of expertise, depending on the specific N-HSOPSC 2.0 factor.</p> Conclusion <p>This study underscores the pivotal influence of organisational and societal conditions on the development of patient safety culture in operating theatre settings. It also highlights the necessity for tailored initiatives to address the unique needs of different fields of expertise. Further research is recommended to explore the underlying mechanisms that influence the patient safety culture in operating theatres.</p>