Nickel-titanium (NiTi) file separation during endodontic treatment is an undesirable event. This phenomenon needs to be understood by knowing the factors influencing fracture in endodontic files. There is a large amount of literature where these factors and their influence have been studied, increasing the knowledge about the mechanisms involved, mainly related to wire technology, file shapes and geometry, operator manipulation, the anatomy of the root canal, and the irrigation and sterilization processes. As many factors are involved, the complexity of the fracture phenomena increases and the isolated correlation of one factor with the file fracture becomes a small part of comprehending the separation phenomena. This thematic review aims to compile important reports from 2014 to 2022 on the factors influencing NiTi file separation. The information obtained was classified into wire technology, file geometry, operational aspects, irrigation and sterilization, and anatomy. For this purpose, the Scopus, Web of Science and ScienceDirect databases were consulted using a search string. Filters were applied to consolidate the final set of relevant papers covering the subject of factors influencing endodontic file separation. It was found that the fracture of NiTi files incorporates different mechanisms that operate simultaneously during the endodontic procedure and strongly affect the instrument performance. The collected information promotes good practices to prevent file separation.