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This study aimed to evaluate and compare canal transportation and centering ability of 4 different root canal preparation systems produced with thermal treatments by means of micro–computed tomographic imaging.Eighty mesial canals of human extracted mandibular molars were selected based on similar morphologic parameters and were randomly assigned to 4 experimental groups (n = 20) according to the canal instrumentation technique: HyFlex CM (HCM [Coltène-Whaledent, Allstätten, Switzerland]), HyFlex EDM (HEDM [Coltène-Whaledent]), WaveOne Gold (WOG [Dentsply Sirona, Ballaigues, Switzerland]), and OneCurve (OC [Micro-Mega, Besancon, France]). The specimens were scanned before and after root canal preparation using X-ray micro-computed tomographic imaging at a resolution of 19.9 μm. Apical transportation and centering ability were then analyzed at 3 different levels: 3 mm, 5 mm, and 7 mm from the apex, representing the apical, midroot, and coronal thirds of the root, respectively. One-way analysis of variance and Kruskal-Wallis tests were used to statistically compare the groups. The significance level was set at 5%.HCM caused less canal transportation than WOG at the 3-mm level in both the buccal and lingual canals (P < .05). Also, HCM resulted in less canal transportation than WOG and OC at the 7-mm level regarding lingual canals. No statistically significant differences were recorded between the groups when the mean centering ratios were compared.The 4 evaluated systems safely prepared root canals causing minimal canal transportation and producing relatively centered preparations. In terms of canal transportation, HCM performed better than WOG at the apical level and better than WOG and OC at the coronal level.