Background: Artificial intelligence (AI) holds great promise in transforming healthcare delivery. However, successful implementation of AI projects in healthcare depends on patients' acceptance and trust. There is only limited empirical research examining public perceptions, particularly on the use of personal health data in AI applications in healthcare. Objective: To examine public knowledge and comfort levels with AI use in healthcare, including use of personal health data with and without consent, and to assess how sociodemographic factors, digital literacy, and health conditions influence these perceptions. Methods: We analyzed data from 6,904 Canadian adults who participated in the 2023 Canadian Digital Health Survey. AI-related knowledge and comfort levels were measured using ordinal scales. Sociodemographic characteristics, digital health literacy, and self-reported chronic health conditions were included as predictors. Ordinal logistic regression models were used to assess associations between these factors and AI-related attitudes. Results: 42.3% reported moderate knowledge of AI, while only 7.8% described themselves as very knowledgeable. Overall, 44.6% were comfortable with AI use in healthcare, increasing to 64.7% when personal health data were used with consent, but decreasing when used without consent (52.6% uncomfortable). Respondents were most comfortable with AI use for epidemic tracking and workflow management, and less so for clinical tasks. Fully weighted ordinal logistic regression models indicated that men (OR=1.57, p<.001 non-citizens higher-income respondents p those with graduate education higher digital health literacy and more chronic conditions exhibited greater odds of reporting ai knowledge. for comfort use in healthcare aged men .001 or comfort. lower-income white reported lower levels. using personal data consent adults were less comfortable than showed while other racial groups without contrast black conclusions: the findings point to enhancing transparent policies ethical governance as key increasing public trust ai-driven healthcare.>