Tuning fork tests, particularly the Rinne and Weber tests, are fundamental in assessing hearing loss. However, medical students often struggle with key auditory physiology concepts, such as the auditory masking effect and sound lateralization. This study evaluated a simulation-based teaching method to enhance first-year medical students' understanding of these concepts. A prospective educational intervention was conducted with 123 first-year Bachelor of Medicine, Bachelor of Surgery (MBBS) students using a pretest and posttest design with a validated questionnaire. During laboratory sessions, students performed tuning fork tests on peers while instructors demonstrated a novel method to simulate unilateral conductive deafness by closing the external auditory meatus with a finger. Students replicated this method before completing posttest evaluations. Pretest data showed that 66.66% of students were uncertain or disagreed with their understanding of the auditory masking effect, and 60.16% were unsure about sound lateralization. Posttest results demonstrated significant improvement: 79.67% strongly agreed they understood the auditory masking effect, and 91.05% strongly agreed they comprehended sound lateralization (P < 0.001). Additionally, student engagement increased, with 96.74% reporting they enjoyed the session compared to 26.01% before the intervention. This study demonstrates that a simple, reproducible simulation-based approach enhances medical students' understanding of auditory physiology concepts. By actively experiencing a simulated unilateral conduction defect, students gain deeper conceptual insight and greater confidence in performing tuning fork tests. This innovative method holds promise for broader adoption in medical education.