Background: Junior colleges have been establishing plans for the development of start-up education for students and have been providing various educational programs to support students’ start-up businesses. Objectives: The effects of entrepreneurship and entrepreneurial self-efficacy on the start-up intention of college physical therapy students were analyzed. Design: Questionnaire design. Methods: Surveys on entrepreneurship, entrepreneurial self-efficacy, and start-up intention were conducted with 269 college physical therapy students, and the levels of individual measurement variables, correlations, and variables affecting start-up intention were analyzed. Results: College physical therapy students showed differences in start-up intention between genders, and male students tended to have a higher startup intention (P<.001) than female students. The correlations among the major variables were analyzed. According to the results, all sub-factors of start-up intention had significant positive correlations with all sub-factors of entrepreneurship and entrepreneurial self-efficacy (P<.05). In addition, multiple regression analysis was conducted to identify factors affecting start-up intention. Start-up education was shown to have no direct effect on start-up intention. Innovation (β=.238, P=.000), risk sensitivity (β=.228, P=.000), and initiative (β=.220, P=.001), which are sub-factors of entrepreneurship, were shown to have effects on start-up intention. Self-regulated efficacy (β=-.193, P=.039) and task challenge (β=.210, P=.004), which are sub-factors of entrepreneurial self-efficacy, were found to have effects on start-up intention. Conclusions: To boost the start-up intention of college physical therapy students and induce their start-ups, customized start-up education that considers entrepreneurship level and gender is necessary.