Background: A needle stick injury is a serious occupational health hazard in health care settings. Health care workers are at risk of bloodborne diseases and the psychological consequences of these injuries. Objectives: we aimed in this study to assess the prevalence of NSI among health care workers in King Saud Medical City and to assess their knowledge, attitude and practices regarding needle stick injuries. Methods: A cross-sectional study was conducted from 17th August 2022 until 30th November 2023. A structured questionnaire from Saudi Ministry of Health guidelines was used to collect the data. The content validity was evaluated by four academics from King Khalid University in Abha, and some minor changes were made. Results: As regard the prevalence of NSI, out of 220 participants, 132 individuals did not report any needle-stick injuries (NSI). While 92.3% of participants accurately defined NSI and acknowledged blood-borne disease transmission risks, only 39.1% followed preventive measures like recapping needles and proper disposal. There were knowledge gaps observed, with varying awareness levels on Hepatitis B and C prevention. Participants demonstrated awareness of post-exposure guidelines but lacked knowledge on immediate actions to take post-injury. Attitudes towards NSI varied, with a significant percentage 44.1% prioritizing patient care over healthcare worker safety. Interestingly, significant support was shown for immediate reporting of sharp injuries and the belief that NSIs are preventable. In terms of practice, adherence to safe needle practices varied, with the majority 90.9% using assigned disposal containers and being vaccinated against Hepatitis B, but not all receiving recent training on safe devices. Conclusion: This study revealed that NSIs remain a prevalent occupational health hazard among healthcare workers. Although a significant number of participants demonstrated adequate knowledge and awareness of NSIs and bloodborne disease transmission risks, there were noticeable gaps in their implementation of preventive measures and immediate post-injury actions. Attitudes towards NSIs varied, with some prioritizing patient care over healthcare worker safety. However, there was strong support for the immediate reporting of sharp injuries and belief in the preventability of NSIs.