BACKGROUND: Artificial Intelligence (AI) Chatbots (e.g., ChatGPT, Microsoft Bing, and Google Bard) can emulate human interaction and may support physiotherapy education. Despite growing interest, physiotherapy students' perspectives remain unexplored. This study investigated Italian physiotherapy students' knowledge, use, and perception of the benefits and limitations of AI Chatbots. METHODS: A cross-sectional study was conducted through Survey Monkey from February to June 2024. One thousand five hundred and thirty-one physiotherapy students from 10 universities were involved. The survey consisted of 23 questions investigating: (a) respondent characteristics, (b) AI Chatbot knowledge and use, (c) perceived benefits, and (d) limitations. Multiple-choice and Likert-scale-based questions were adopted. Factors associated with knowledge, use, and perceptions of AI were explored using logistic regression models. RESULTS: Of 589 students (38%) that completed the survey, most were male (n = 317; 53.8%) with a mean age of 22 years (SD = 3.88). Nearly all (n = 561; 95.3%) had heard of AI Chatbots, but 53.7% (n = 316) never used these tools for academic purposes. Among users, learning support was the most common purpose (n = 187; 31.8%), while only 9.9% (n = 58) declared Chatbot use during internships. Students agreed that Chatbots have limitations in performing complex tasks and may generate inaccurate results (median = 3 out of 4). However, they neither agreed nor disagreed about Chatbots' impact on academic performance, emotional intelligence, bias, and fairness (median = 2 out of 4). The students agreed to identify the risk of misinformation as a primary barrier (median = 3 out of 4). In contrast, they neither agreed nor disagreed on content validity, plagiarism, privacy, and impacts on critical thinking and creativity (median = 2 out of 4). Young students had 11% more odds of being familiar with Chatbots than older students (OR = 0.89; 95%CI 0.84-0.95; p = < 0.01), whereas female students had 39% lesser odds than males to have used Chatbots for academic purposes (OR = 0.61; 95%CI 0.44-0.85; p = < 0.01). CONCLUSIONS: While most students recognize the potential of AI Chatbots, they express caution about their use in academia. Targeted training for students and faculty, supported by institutional and national guidelines, could guarantee a responsible integration of these technologies into physiotherapy education. TRIAL REGISTRATION: Not applicable.