Background: Safety culture is increasingly recognized as an important strategy to improve patient safety. Objectives: The present study aimed at assessing workforce perception of hospital safety culture at Shark El Madina hospital and to identify differences in perception of safety culture dimensions among different professional categories at the study hospital. Methods: The study utilized a self administered questionnaire titled “Patient Safety Climate in Health Care Organizations" (PSCHO) which is composed of 38 items. A total of 186 completed questionnaires were collected distributed among 64 physicians, 74 nurses and 48 technicians. The mean score of items and dimensions were calculated as well as the percent problematic response (PPR) which isthe fraction of survey participants who answered in ways that are inconsistent with an optimal safety culture. Results: The study revealed that the overall PPR was 46.9%, being highest among nurses (53.1%) %), followed by physicians (44.3%) and lowest among technicians (39.5%). Nurses had higher percentage of PPR than other professional groups in 8 dimensions. The 3 dimensions which had the lowest mean scores among the three professional categories were fear of blame followed by unit recognition and support for safety followed by organizational resources for patient safety. Nurses showed lower mean scores in five out of nine safety culture dimensions. Conclusion and Recommendations: Patient safety culture among health care workers at Shark El Madina hospital is relatively negative. There are differences among professional categories with nurses showing more negative view of safety culture dimensions. It is recommended to establish safety cultural change through education and training of staff, encouragement of no blame environment and provision of resources for patient safety.