Dermestid beetles (Coleoptera: Dermestidae) are necrophagous insects feeding on mummified carcasses. After six to seven molts, the larvae stop feeding and dig pupation chambers to hide and safely evolve into adults. Such pupation chambers have already been observed on archaeological mammals’ bones, but the attribution and interpretation of these osteological lesions lack experimental evidence in a forensic context. To observe whether dermestid larvae dig pupation chambers in human bones, 20 or 40 Dermestes maculatus (De Geer, 1774) larvae were placed in a dermestarium with different types of bones varying in species (Bos taurus or human), age (adult or immature), and preservation method (fresh or dry). Our results show that dermestid larvae caused multiple lesions, including larval mandible traces on cortical bone, cortical perforations, drilling of pupation chambers, destruction of the trabecular network, and the perforation of cartilage. Bone destruction was mainly observed on aged dry bones, while fresh bones only exhibited soft tissue and superficial cartilage lesions. According to these results, pupation chambers could indicate the simultaneous presence of several corpses at different decomposition stages, or the addition of new corpses while others were already skeletonized. These conclusions are particularly important in the case of mass graves, where chronology is sometimes difficult to establish.