BACKGROUND: Artificial intelligence (AI) is transforming healthcare by enhancing diagnostics, treatment planning, and patient management. However, the successful integration of AI depends on healthcare professionals' (HCPs) knowledge, attitudes, and practices (KAP). The study aimed to evaluate the KAP of HCPs regarding the role of AI in healthcare. Additionally, the study aimed to identify key barriers, such as ethical concerns and technical limitations, that hinder AI adoption. MATERIALS AND METHODS: A cross-sectional study was conducted using an online structured questionnaire targeting doctors, medical students, and nurses. The survey, distributed Google Forms, measured participants' demographics, KAPs regarding AI. Purposive and snowball sampling methods were employed to recruit a total of 350 participants. RESULTS: A total of 350 participants completed the survey, with the majority being medical students (52%) and younger professionals aged 18-27 years (73.1%). AI's primary use was in diagnostics (71.1%), followed by treatment protocol development (45.4%) and drug development (33.1%). Most respondents rated their technical skills as low or very low, with medical students showing slightly higher proficiency compared to nurses and faculty members. Ethical concerns were prominent, with 46% of participants highlighting data privacy and security issues, followed by concerns about high costs (23.7%) and over-reliance on manual processes (16.6%). Significant differences were observed between professions: medical students and faculty were more open to AI's role in making critical decisions (60.98% and 61.94%, respectively) than nurses (29.41%). Additionally, 97% of nurses expressed willingness to undergo AI training compared to 86% of faculty and 82% of medical students. However, confidence in AI's ability to improve clinical outcomes was mixed, with 35.71% agreeing, 29.71% neutral, and 40% regularly using AI tools in practice. CONCLUSION: The findings highlight a positive attitude toward AI in healthcare, with substantial recognition of its benefits, especially in diagnostics. However, significant gaps in knowledge, technical skills, and awareness of ethical issues remain, particularly among nurses. Tailored education and training programs are needed to bridge these gaps and enhance AI readiness in healthcare practice.