Middle ear ossicles are critical for audition and rarely preserved in fossils. Based on microcomputed tomography images, our comparative 3D shape analysis of Neandertal ossicles shows striking shape differences between Neandertals and anatomically modern humans (AMHs). However, these morphological differences do not affect the functional properties of the ossicles, potentially indicating consistent aspects of vocal communication in Neandertals and AMHs. Instead, a strong relationship between ossicle morphology and tympanic cavity architecture is found.