© 2022 Japanese Stomatological Society.Aim: In this cross-sectional descriptive study, it was aimed to determine the relationship between the prevalence of temporomandibular disorder (TMD) and oral parafunctions in dental assistant students and to evaluate the effects of gender and education years. Methods: The results of 105 individuals (mean age: 20.72 ± 3.51 years; 75 females, 30 males) were analyzed in this descriptive cross-sectional study, which included student participants who received dental assistant education program from Erciyes University Vocational Health College. The Fonseca Anamnestic Index (FAI) questionnaire was used to define the presence and symptoms of TMD, and the Oral Behavior Checklist (OBC) questionnaire was used to determine the oral parafunctions of individuals. Fisher's exact, Mann–Whitney U, Kendall tau-b, and Spearman correlation and linear regression analyses were used to evaluate the data. Results: Any level of TMD was detected in 64.8% of all individuals included in the study, and most of these patients were women (74.3%). No significant difference was found between the TMD prevalence of the first- and second-grade students based on the years of education of the participants (p = 0.614). Considering the sex and class parameters, statistically significant weak (r = 0.378 for females; r = 0.375 for the first grade) and middle (r = 0.418 for males; r = 0.478 for the second grade) degree positive correlation was determined (p < 0.01) between the OBC and TMD scores for both parameters. Conclusion: The prevalence of TMD is higher in dental assistant students, especially females, and this is associated with certain degrees of oral parafunctions.