Background: Safety culture is a subset of organizational culture, and beliefs and values specifically reflect health and safety. The present study aimed to measure patient safety culture in a selected hospital with a national accreditation approach and safety-friendly hospital standards. Methods: The present study is a cross-sectional study conducted in 2018-2019 at Baqiyatallah University of Medical Sciences. This applied research is a descriptive study conducted using a qualitative method. The sample size of this study was determined using the Richard J. Shavelson formula. Then, 262 hospital staff were selected, of whom 257 participated in the first stage and 201 in the second stage. The Hospital Survey on Patient Safety Culture (HSOPSC) questionnaire was used to assess safety culture. Results: Comparing the first stage of monitoring safety culture with the second stage of interventions showed that the lowest level of safety culture was associated with the number of reported incidents, and the patient safety grade had the highest rank. Also, the difference between the first and second stage monitoring was shown in determining the level of safety culture, workplace (+0.3), direct manager (-0.1), communication and feedback (+0.1), frequency of reported incidents (-0.1), patient safety level (+0.2), your hospital (-0.1), number of reported incidents (+0.3), and their relationship was not significant. Conclusions: According to the results of this study, it is possible to promote an effective safety culture by strengthening interdepartmental cooperation and strengthening communication between hospital units through interaction and intimate communication between supervisors and subordinates.