It is stated that engine-driven instruments might cause dentinal defects during root canal preparation. These defects might spread and progress into greater fractures or vertical root fracture. This study aimed to compare the incidence of dentinal defects that might arise all through preparation of root canals, using One Shape, RaCe, and WaveOne systems in canals with a curvature (0-20°) and (20-40°). In this in vitro study, 150 mandibular first molars were enrolled. Based on the degree of curvature in the mesial roots, the samples were divided into two groups (n=75) of straight and moderately curved canals (0-20º), and severely curved canals (20-40°). Then each group was randomly divided into four sub-groups. In subgroups 1 to 3 from each group, canals were prepared using WaveOne, One Shape, and RaCe. Then all roots were sectioned at 3, 6 and 9-mm distances from the apex. The slices were evaluated using stereomicroscope at 12× magnification. The data were analyzed using the Chi-square and Fisher's exact tests and the level of significance was set at 0.05. Fracture and other defects were not found in the control groups. In canals with curvature (0-20°), WaveOne caused the maximum dentinal defects and RaCe produced the least. Moreover, in canals with curvature (20-40°), One Shape caused the maximum dentinal defects while WaveOne and RaCe caused equal dentinal defects approximately. There was a statistically significant relationship between the performance of RaCe and One Shape in canals with curvature (0-20º) and (20-40º), (pValue< 0.05) while no statistically significant difference was observed in connection to the performance of WaveOne (p> 0.05).