Neuroanatomy education is challenging due to the spatial complexity of brain structures and the difficulty of three-dimensional visualization in traditional teaching methods. Immersive learning technologies, such as 3D Virtual Learning Objects (VLOs), have demonstrated potential for improving knowledge comprehension in advanced educational settings, aligning with the principles of Smart Education and Digital Twins. This research seeks to evaluate the impact of 3D VLOs as digital twins of neuroanatomical structures on improving the spatial understanding of neuroanatomy students, promoting immersive and active learning strategies.An interactive 3D VLO modeling key brain structures was implemented, integrated into a virtual learning platform. The methodology used was ADDIE for the development of the VLOs and the educational process. An experimental design was applied. Spatial understanding and retention were measured pre- and post-intervention, complemented by usability and learning perception surveys using the TAM model. The hypothesis was tested using the Wilcoxon test, and the alternative hypothesis that VLOs improve spatial understanding of neuroanatomy was accepted. The results identified that students who used 3D OVAs significantly improved their spatial understanding and retention of neuroanatomical content compared to traditional methods. They also reported greater motivation, satisfaction, and interactivity with learning. 3D OVAs as digital twins constitute an effective tool for teaching neuroanatomy and, in Education 4.0, enhancing spatial understanding. This strategy aligns with the objectives of Smart Education and the integration of advanced learning technologies, offering a replicable model for education in health sciences and other disciplines that require complex spatial visualization.