Introduction Knowledge and attitudes of medical professionals and students towards artificial intelligence in the context of medical research remain poorly explored. Objective To determine the knowledge and attitudes about artificial intelligence in Cuban health professionals and students. Methods Cross-sectional descriptive study. A validated survey was used to gather data on demographics, AI resources, perceived knowledge, attitudes, confidence, and interest in AI policy. Ordinal regression was conducted to explore the influence of participant roles on attitudes and knowledge about AI. Results This study surveyed 728 Cuban health professionals and students, revealing a moderate level of AI knowledge, with significant differences between groups. Social networks were the primary source of AI information, Academics-Researchers also relied on academic journals and textbooks. Confidence in assessing AI outputs and mitigating clinical harms was moderate, but a majority (45.05%) believed AI could change their roles. Interest in reviewing or developing AI policies was high (39.97%). Undergraduate students expressed greater interest in learning about AI applications and confidence in understanding machine learning, while Academics-Researchers demonstrated strong attitudes towards AI and interest in policy development. A significant barrier to AI learning across groups was a lack of institutional support, though lack of time was also prevalent among undergraduate students. Conclusions Cuban health professionals and students demonstrate moderate AI knowledge, with significant differences between groups. Interest in AI and its potential impact on their roles is high, but barriers to adoption, like lack of institutional support and time, require attention.