Objectives: This study aims to compare the cyclic fatigue resistance of VDW.ROTATE, TruNatomy, 2Shape, and HyFlex CM nickel-titanium (NiTi) rotary files at body temperature. Materials and Methods: In total, 80 VDW.ROTATE (25/0.04), TruNatomy (26/0.04), 2Shape (25/0.04), and HyFlex CM (25/0.04) NiTi rotary files (n = 20 in each group) were subjected to static cyclic fatigue testing at body temperature (37°C) in stainless-steel artificial canals prepared according to the size and taper of the instruments until fracture occurred. The number of cycles to fracture (NCF) was calculated, and the lengths of the fractured fragments were measured. The data were statistically analyzed using a 1-way analysis of variance and post hoc Tamhane tests at the 5% significance level (p < 0.05). Results: There were significant differences in the cyclic fatigue resistance among the groups (p < 0.05), with the highest to lowest NCF values of the files as follows: VDW.ROTATE, HyFlex CM, 2Shape, and TruNatomy. There was no significant difference in the lengths of the fractured fragments among the groups. The scanning electron microscope images of the files revealed typical characteristics of fracture due to cyclic fatigue. Conclusions: The VDW.ROTATE files had the highest cyclic fatigue resistance, and the TruNatomy and 2Shape files had the lowest cyclic fatigue resistance in artificial canals at body temperature.