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AIM: To determine the incidence of ProFile nickel-titanium rotary instrument fracture in an endodontic laboratory simulation. METHODOLOGY: Three hundred and sixty dental students used 2880 ProFile nickel-titanium rotary instruments to prepare 1440 simulated root canals in 720 plastic teeth, and another 2880 ProFile nickel-titanium rotary instruments to prepare 1440 natural root canals in 720 extracted teeth. A standardized crown-down rotary instrumentation technique was used, which included measures to prevent fracture. Rotary instrument fracture was monitored during and after completion of the laboratory simulation exercises to determine the incidence of fracture. When fracture occurred, data were collected concerning the size of the instrument, the length of the instrument fragment, the location of the fragment in the canal and the curvature of the canal in which the instrument fractured. RESULTS: The incidence of instrument fracture was 0.41% in plastic simulated canals and 0.31% in natural root canals. The overall incidence of instrument fracture was 0.36%. Of the instruments that fractured 67% were size 25, 0.04 taper; and 81% of the fragments were located in the apical third of the canal. The mean, median and mode of the fragment lengths were all 3 mm. CONCLUSIONS: The low overall incidence of fracture in this study suggests that ProFile rotary instruments are safe for use by dental students in laboratory simulations and that if preventive measures are taken the incidence of instrument fracture can be minimized