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This study aimed to evaluate and compare the frequency of microcracks and its severity at different root canal dentin locations, after preparation with thermomechanically heat-treated engine-driven nickel-titanium instruments. Forty mandibular premolars were assigned to four experimental groups (n = 1 ProTaper Next, ProTaper Gold, WaveOne Gold and Reciproc Blue. After pre-instrumentation micro-computed tomography scans, the root canals were prepared to size 25. Following post-instrumentation scans, pre- and post-instrumentation scann