BackgroundThe aim of this study was to compare cyclic fatigue resistance (CFR) and separation patterns during rotating movement of five heat-treated nickel titanium files in both single and double-curved canals. MethodsCFR was compared between five groups of 24 files each: HyFlex® EDM, HyFlex™ CM, Vortex Blue®, Protaper Next®, and One Curve, by determining the time needed to fracture (TTF) and the number of cycles to failure (NCF) in a cyclic fatigue testing device with a single 5-mm radius of curvature (ROC) and 60° angle, and a double curvature; coronal curvature (5 mm ROC and 60° angle) and apical curvature (2 mm ROC and 70° angle). The separation pattern was evaluated, and file fragment separated length were measured. One-way ANOVA and Tukey’s post-hoc were used to compare statistical differences between the groups. Scanning Electron Microscope (SEM) was used to investigate the fracture sites. Results HyFlex™ CM files displayed significantly higher CFR, followed by HyFlex® EDM and Vortex Blue® in single curvature canals, and Protaper Next® (PTN) in the double curvature canals, as observed by TFF and NCF. PTN group had the highest tendency for files to extrude apically after separation in both curvatures. ConclusionHyFlex™ CM instruments have superior resistance to cyclic flexural fatigue within single canals with single and double curvatures, followed by HyFlex® EDM and Vortex Blue® instruments. Protaper Next® files followed by One Curve had the highest tendency to extrude apically after separationClinical relevanceHyFlex™ CM instruments showed the highest safety in preparing canals with single or double curvatures, considering CFR.