Background: Patient safety culture plays a pivotal role in enhancing care quality, pre-venting errors, promoting clinical outcomes, and reducing healthcare costs. It also helps min-imize adverse events, increasing patient safety, and elevating the quality of medical care pro-vided. This study aims to evaluate the perception of patient safety culture within a university hospital. It aims to examine the correlation between the demographic and workplace charac-teristics of the staff and their perceptions of patient safety culture. Methods: A quantitative cross-sectional study design was employed for this study. The Arabic version of the Hospital Survey of Patient Safety Culture (HSOPSC) v2.0 was used to evaluate the perception of pa-tient safety culture among hospital staff. The survey was distributed to all staff via email us-ing the Survey Monkey® online platform over two weeks. Descriptive statistics were used to measure the perception. The relationship between variables was determined using the Mann-Whitney U test, Kruskal-Wallis test, and multiple linear regression. Results: “Staffing and Workplace” (59.5%) and “Response to Error” (51.9%) were found the weakest composites. “Teamwork” (82.5%), and the “Organizational Learning—Continuous Improvement” (81.1%) were the highest positive composites. The rest of the composites were perceived pos-itively. Nursing staff perceived the patient safety culture more positively than other profes-sions (Mean Rank = 157.23, p-value. Conclusions: This study presents an assessment of pa-tient safety culture among healthcare staff at a university hospital. The staff perceived the overall patient safety culture positively. However, efforts must be made to improve the two weak composites. A qualitative approach study could be done to explore deeply the reasons for this weakness in these composites. The demographics of the staff were not associated with patient safety culture.