ObjectivesTo investigate the efficacy of retreatment and reciprocating rotary systems in removing gutta-percha filling material from root-canal systems.MethodsA total of 90 single-canal human teeth were instrumented using a F3 ProTaper file and obturated with gutta-percha and the Tubliseal sealer using the lateral-compaction technique. They were divided into five groups according to the rotary system used to remove the filling material: Reciproc, which used the Reciproc-R25 file; WaveOne, which used the WaveOne-Primary file; S1, which used the S1 (25/.06) file; ProTaper-R, which used the ProTaper-Retreatment system; and Mtwo-R, which used the Mtwo®-Retreatment system. The time required to remove the filling material (min) and the associated instrument separation, if any, were recorded. Teeth were cleaved longitudinally and photographed with a high-resolution camera. Images were imported to the AutoCAD application to measure the remaining filling material and the root-canal space. Data were analysed using the one-way analysis of variance and Chi-squared tests.ResultsThe Mtwo-R group showed the highest mean remaining filling material (51%) (p < 0.001), with no significant differences among the other groups. The ProTaper group required the least time for filling-material removal (4.95 min), with significant differences compared to the other groups (p < 0.05), except the WaveOne group (5.83 min; p = 1.000). Overall, 13 instruments (15.9%) were used for filling-material removal, with a significantly greater proportion in the Mtwo-R group (33.3%) compared to the other groups (p = 0.009).ConclusionsWithin the limitations of this in vitro study, reciprocating rotary systems were as effective as retreatment rotary systems in removing root-canal filling material. The Mtwo-R system showed the poorest performance with respect to removal effectiveness and instrument separation.