Health education is one of the knowledge areas in which augmented reality (AR) technology is widespread, and it has been considered as a facilitator of the learning process. In literature, there are still few studies detailing the role of mobile AR in neuroanatomy. Specifically, for the spinal cord, the teaching-learning process may be hindered due to its abstract nature and the absence of three-dimensional models. In this sense, we implemented a mobile application with AR technology named "NitLabEduca" for studying the spinal cord with an interactive exploration of 3D rotating models in the macroscopic scale, theoretical content of its specificities, animations, and simulations regarding its physiology. To investigate "NitLabEduca's" effects, eighty individuals with and without previous neuroanatomy knowledge were selected and grouped into control and experimental groups. Divided, they performed learning tasks through a questionnaire. We used the System Usability Scale (SUS) to evaluate the usability level of the mobile application and a complimentary survey to verify the adherence level to the use of mobile applications in higher education. As a result, we observed that participants of both groups who started the task with the application and finished with text had more correct results in the test (p < 0.001). SUS results were promising in terms of usability and learning factor. We concluded that studying the spinal cord through "NitLabEduca" seems to favor learning when used as a complement to the printed material.