: Anesthesia staff commonly have a number of incidences of needle stick and sharp injuries (NSIs), due to their work nature, which has emergency and critical cases. The aim of this study to is identify and examine the factors associated with NSIs incidence among anesthesia staff. A cross-sectional online survey was conducted randomly on a sample of Anesthesia staff working in operating theatres from major hospitals in the Eastern region, of Saudi Arabia. Data on demographic characteristics, history of NSIs, nature of work, compliance with infection control protocols, and knowledge of infection control procedures and disease transmission were collected using a self-administered questionnaire. Data were analyzed with demographic variables by using t-test and ANOVA to find the significant difference. A P value of ≤0.05 is considered for statistical significance. Overall (89.2%) of the participants accidentally suffered NSIs, of which 60.2% said this occurred monthly during routine cases, emergencies, or on-calls, and about (35.5%) reported contamination after their injury. The most commonly reported reason for NSIs was during withdrawing drugs of anesthesia from ampules, needle recapping, and placing a needle in a sharp’s container. Overall, participants had inadequate practices regarding standard precautions. NSIs is a common public health problem in operating rooms among anesthesia staff, particularly during emergencies and stressful work schedules. The promotion of decent working conditions, the elimination of excessive use of injections, and the observance of general precautions will be important for the future control of potential blood-borne pathogens infections from occupational exposure to sharps in this environment..