Abstract: Determination of the glide path is recommended before using rotary instruments. This study aimed to evaluate the dynamic cyclic fatigue resistance of new and used glider rotary instruments in up to 6 root canals. Seventy-two TruNatomy Glider files were used for the preparation of root canals of extracted lower molars, which were then submitted to the dynamic cyclic fatigue test carried out in a curved metallic artificial canal. The instruments were divided into 4 groups (n = 18): Control group, new instruments without any use in the root canal; Group 2U, instruments used in 2 mesial canals; Group 4U, instruments used in 4 mesial canals; Group 6U, instruments used in 6 mesial canals. The time to failure (TF) of the instrument was recorded, and the number of cycles to failure (NCF) was calculated. The data were submitted to 1-way analysis of variance and to the Games-Howell test for multiple comparisons, adopting a significance level of 5%. TF and NCF were significantly affected by the number of file uses. The Games-Howell test revealed that TF and NCF were significantly greater in the control group than in Group 4U. In Group 2U, TF and NCF were intermediate and not significantly different from the control group. Group 6U had significantly lower TF and NCF than all other groups. The TruNatomy Glider can be used as a glide path for up to 2 mesial canals of mandibular molars, whereas its use on 4 or 6 root canals is not suggested Ver menos