Objectives: This micro-computed tomographic (CT) study aimed to compare the shaping abilities of ProTaper Next (PTN), One Shape (OS), and One Curve (OC) files in 3-dimensionally (3D)-printed mandibular molars. Materials and Methods: In order to ensure standardization, 3D-printed mandibular molars with a consistent mesiobuccal canal curvature (45°) were used in the present study (n = 18). Specimens were instrumented with the OC, OS, or PTN files. The teeth were scanned preand post-instrumentation using micro-CT to detect changes of the canal volume and surface area, as well as to quantify transportation of the canals after instrumentation. Two-way analysis of variance was used for statistical comparisons. Results: No statistically significant differences were found between the OC and OS groups in the changes of the canal volume and surface area before and after instrumentation (p > 0.05). The OC files showed significantly less transportation than the OS or PTN systems for the apical section (p < 0.05). In a comparison of the systems, similar values were found at the coronal and middle levels, without any significant differences (p > 0.05). Conclusions: These 3 instrumentation systems showed similar shaping abilities, although the OC file achieved a lesser extent of transportation in the apical zone than the OS and PTN files. All 3 file systems were confirmed to be safe for use in mandibular mesial canals.