Dietary supplements are concentrated sources of nutrients used as an addition to a normal diet with a nutritional or physiological effect. In this cross-sectional study, it was aimed to determine the factors of dietary supplement use, the related knowledge, and their relationship with supplement use in a faculty of health sciences among university students. This cross-sectional study was carried out with 333 voluntary university students from Ankara Yıldırım Beyazıt University, the Faculty of Health Sciences. A questionnaire form consisting of 5 sections was applied to the students by face-to-face interview method. The most remarked purposes of supplement use were to improve health (44.2%) and boost immunity (22.1%). There was no difference between age, gender, department, coexistent disease, tobacco smoking and alcohol consumption in terms of dietary supplement use (p = 0.611, p = 0.720, p = 0.682, p = 0.062, p = 0.491, and p = 0.658, respectively). No significant difference was observed between physical activity status, the duration of physical activity, dieting status, the source of diet program, daily frequency of main meals and snacks, body mass index (BMI) groups, and the reason for meal skipping among students according to dietary supplement use (p = 0.180, p = 0.205, p = 0.686, p = 0.226, p = 0.533, p = 0.449, p = 0.976, and p = 0.758, respectively). There was not any significant difference between physical activity status, the duration of physical activity, dietary supplement use, dieting status, the frequency of main meals and snacks and BMI in terms of total knowledge score of dietary supplements (p = 0.245, p = 0.713, p = 0.278, p = 0.067, p = 0.466, p = 0.647, and p = 0.851, respectively). Scientific attention should be paid to the use of dietary supplements. Arbitrary approaches can lead to unwanted interactions. Therefore, in order for health and government policies to be established related to dietary supplements, it is essential to determine the factors related to their use.