Background: Fulfilling aesthetic norms is an imperative of modern society. Accordingly, aesthetics has found its important role in dental medicine. The aim was to study whether there is a difference in the perception of tooth shade change and anatomical variations of anterior maxillary teeth among dental students depending on the level of education, gender, and experience in assisting in a dental office. Methods: The research was conducted among dental students via the Google Forms platform. Students evaluated the aesthetic acceptability of a smile on the unaltered, reference photograph (RP) and on the other 28 photographs in which the tooth shade (lighter and darker teeth), length (shortened central incisors, elongated, and beveled lateral incisors, elongated canines) and position (atypical and typical rotations and diastemas) of the upper maxillary teeth were altered by digital manipulation. The 1-10 assessment scale was used. Statistical analysis comprised one-way Kolmogorov-Smirnov test, X-2 tests, t-test, and 3-way MANOVA. Results: In the research 208 students participated, 113 were preclinical students, 104 assisted in dental office and 175 were females. There were no significant effects of gender regarding length and position modifications (p > .05), while women were stricter in evaluation of lighter shade (p < .05). Clinical students were stricter in evaluating all manipulated photographs (MP) except those towards darker shade modifications, while assisting in a dental office or having a dentist in close family showed no significant effects (p > .05). Conclusions: With a higher level of targeted academic education, dental students sharpen their ability to notice deviations from the aesthetic norms of a smile.