Background <p>The healthcare sector is undergoing a digital transformation, where the integration of Artificial Intelligence (AI) plays a vital role in reshaping healthcare practices. AI technologies promise to improve work procedures, mitigate future risks, and expedite proactive patient care. This guide seeks to address the significant knowledge gap among healthcare professionals, focusing on AI’s integration within healthcare systems and its impact on patient safety culture.</p> Methods <p>A descriptive correlational design was implemented to explore the correlations between healthcare providers’ knowledge and attitudes toward artificial intelligence (AI) and their perception of patient safety culture. A non-probability, convenient sample of 238 healthcare providers comprising physicians, nurses, and technicians working at Najran University Hospital was included in the study. Data collection was conducted using two self-reported questionnaires: the Structured Knowledge and Attitude Questionnaire on Artificial Intelligence and the Hospital Survey on Patient Safety Culture (HSOPSC).</p> Results <p>The research findings indicated that the healthcare providers’ knowledge and attitude toward AI are significant predictors of patient safety culture, with a combined explanatory power of 60% (<i>R</i>² = 0.60). Both knowledge (B = 0.296, <i>p</i> = .000) and attitude (B = 0.502, <i>p</i> = .000) show significant positive relationships with patient safety culture, indicating that higher knowledge and a more positive attitude toward AI contribute to a stronger patient safety culture.</p> Conclusions <p>This study's findings emphasize the need to enhance healthcare providers’ knowledge and attitudes toward AI to reinforce a culture of patient safety. Healthcare institutions are encouraged to incorporate AI-centered education and training programs to enhance providers’ comprehension and confidence in leveraging AI for safer clinical practice.</p>