As a vital part of patient care delivery, patient safety culture contributes to the quality of care provided by nurses. Safe patient care is positively linked to the attitudes of nurses. This study aimed to assess the perception of nurses working in a newly established teaching hospital. A cross-sectional study involving 194 nurses from three different units was conducted by using a 24-item Hospital Survey of Patient Safety Culture. Data on gender, working unit, age, years of working, and attendance in workshops on patient safety were also collected. The majority of the nurses had a positive total score of patient safety culture. The lowest score was 76 (63%), and the highest score was 120 (96%). The awareness on patient safety culture significantly differed between gender, years of working, and working units. Post-hoc comparisons using Tukey’s HSD test yielded a significant difference between nurses from critical care units and those from medical and surgical units. The mean score and total positive score on awareness on patient safety culture of the former were higher than those of the latter. Overall, the majority of the staff nurses in International Islamic University Malaysia Medical Center had a positive total score on awareness on patient safety culture. Awareness on patient safety, which is considered crucial worldwide, should be enhanced to influence the development of a positive patient safety culture within hospitals. This implementation would directly develop high-quality care to patients and positively impact health organizations.