Introduction <p>Despite its critical role in esthetics and function, dental anatomy receives less attention than tooth color during dental education.</p> Aim and objectives <p>This study aimed to assess the impact of dental anatomy training on the esthetic beliefs and opinions of dental students.</p> Method and materials <p>A cross-sectional survey was conducted among 1st-year(<i>n</i> = 163) and 6th-year( <i>n</i> = 143) dental students at the Faculty of Dental Medicine, Tunisia. The questionnaire included 7 dichotomous yes/no questions and one multiple-choice question. It was validated by experts and then sent via Google Form to the targeted students. Statistical analysis was performed using SPSS software. Significance level was set at 5%.</p> Results <p>Almost all students agreed that the smile affects social life and that tooth morphology impacts the esthetics 42.3% of 1st year students and 93% of interns agreed that tooth morphology varies between genders, while 65% of first-year students and 86.7% of interns agreed that some features vary among ethnic. Almost all agree that teeth carving helps to understand the teeth morphology (84.0% and 76.2% respectively for first-year students and interns). Only 25.2% of 1st year Students were familiar with the term of the Golden Proportions. Almost all students agreed that the central incisor is the most involved tooth in esthetics. Statistically significant differences were reported according to sex.</p> Conclusion <p>Close agreement between 1st year students and interns suggests the impact of dental anatomy training. Thus, dental anatomy significantly influences students’ esthetic perception.</p>