IntroductionThe aim of this study was to compare the shaping ability of four root canal preparation systems in newly developed 3D-printed root canal models.Materials and methodsFor this study, 1080 3D-printed acrylic resin blocks with nine different root canal configurations were produced. They were prepared with Reciproc R25 (#25), F6 SkyTaper (#25 and #30) F360 (#25 and #35) and One Shape (#25) (N = 30 per system). Pre- and post-instrumentation images were superimposed for evaluation of the centering ratio of the different systems. Ledges, instrument fractures and preparation times were also recorded. Analysis of variance (ANOVA) and post-hoc Tukey tests were conducted, comparing the mean canal centering ratios and the mean preparation times.ResultsThere were significant differences between all systems regarding the centering ratios in the different root canal configurations (ANOVA p < 0.001). The root canal configuration had considerable effect on the centering ratio of the instruments. The best overall mean centering ratios were achieved with F6 SkyTaper #25 instruments especially in canal configurations with big curvature angles and radii, while F360 #35 was least centered especially in canals with small curvature angles and radii. Most ledges occurred with OneShape, while it was the significantly (p < 0.001) fastest preparation system (86.7 s (SD 13.53)) and Reciproc the significantly (p < 0.001) slowest (103.0 s (SD 20.67)).Conclusion3D-printed root canals are suitable to produce challenging canal configurations and to investigate the limitations of root canal instruments. We found that all instruments caused canal transportations. However, F6 SkyTaper #25 files had better overall centering ratios than the other instruments. In canal configurations with small curvature radii, the centering ratio of some instruments is low and the probability for ledges is increased.