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The purpose of this study was to evaluate the effect of different temperatures (0°C, 10°C, 22°C, 37°C, and 60°C) on the cyclic fatigue life of nickel-titanium (NiTi) files using a new fatigue test model in zirconium oxide.Three superelastic NiTi files (EndoSequence [Brasseler USA, Savannah, GA], ProFile [Dentsply Tulsa Dental Specialties, Tulsa, OK], and K3 [SybronEndo, Orange, CA]), and 3 heat-treated (K3XF [SybronEndo], Vortex [Dentsply Tulsa Dental Specialties], and HyFlex CM [Coltene-Whaledent, Allstetten, Switzerland]) NiTi files, all size 25/.04, were subjected to cyclic fatigue tests inside a novel, artificial ceramic canal with a curvature of 60° and a 5-mm radius. The model was immersed in water at 5 different preset temperatures. The number of cycles to failure (NCF) was recorded, and the fracture surface of the fragments was examined by a scanning electron microscope. The data were analyzed using 2-way analysis of variance with the significance level at 0.05.When the temperature was reduced from 60°C to 0°C, the NCF significantly increased from over 2 to 10 times for the NiTi file groups (P < .01). K3XF had the highest fatigue resistance of all files at 0°C (P < .05). Vortex files had the highest NCF at 10°C–60°C. The NCF of heat-treated files was significantly higher than superelastic NiTi files at 10°C and 20°C (P < .05). There was no significant difference in the NCF of HyFlex CM at 0°C and 22°C. There was little difference in the fractographic appearance among different temperatures, except that the fraction area occupied by the dimple region of some instruments at 0°C was slightly smaller than at 60°C.Cooling down to low temperatures may be an interesting strategy to improve the fatigue resistance of rotary NiTi files.The effect of different temperatures (0°C, 10°C, 22°C, 37°C, and 60°C) on the cyclic fatigue life of nickel-titanium (NiTi) files is evaluated using a new fatigue test model in zirconium oxide.When the temperature is reduced from 60°C to 0°C, the number of cycles to failure (NCF) significantly increases from over 2 to 10 times for the NiTi file groups.K3XF has the highest fatigue resistance of all files at 0°C.The Vortex file has the highest NCF at 10°C–60°C.The NCF of heat-treated files is significantly higher than superelastic NiTi files at 10°C and 20°C.