Objectives: The aim of this cross-sectional study was to evaluate the level of knowledge about oral health and oral hygiene habits among the student population of the University of Split. Materials and methods: The research was based on an online survey questionnaire, and was conducted among 1088 students, 32.4% of which study in the field of biomedicine and health. Data were processed by Mann-Whitney or Kruskal Wallis 1-way ANOVA test. Statistical significance was set at P ≤ 0.05. Results: It was confirmed that the respondents have a good level of oral health knowledge according to the median answer 11 (9-13) for all surveyed students, where 4.3% of the respondents answered all the questions correctly. Respondents of older age (≥ 23 years) show better knowledge of oral health than the younger ones (P ≤ 0.001), as well as respondents of higher years of study (4 - 6 years) compared to those of lower age (P ≤ 0.001), and also students attending one of the biomedical studies (P ≤ 0.001). Most respondents brush their teeth with a toothbrush and toothpaste several times a day (85.7%) between two and three minutes (70.5%). Only 26.7% of respondents use dental floss daily, and 15.3% use interdental brushes. Conclusions: Students of the University of Split have enough knowledge about oral health, but most of them visit their dentist only when needed. Oral-hygiene habits of the respondents showed that a small number of them use additional means such as dental floss or interdental brushes. The obtained data emphasize the importance of additional education of students with the aim of better understanding and maintaining oral health, and thus overall health.