BACKGROUND This study evaluated the effect of traditional and conservative endodontic access cavity designs in combination with WaveOne Gold and TruNatomy instrumentation systems on the fracture resistance of mandibular first molars by means of nonlinear finite element analysis (FEA). METHODS Micro-CT images of 4 human mandibular first molars were used to generate representative FEA models. The mandibular first molars samples were scanned before and after endodontic access cavity preparation and instrumentation of all 3 canals. Five nonlinear static loads were applied vertically and horizontally to specific contact points on the occlusal surface of the teeth. Maximum von Mises stress before failure and distribution of von Mises strains were recorded and compared between groups. RESULTS Molars with conservative endodontic access cavities required similar levels of loads to reach failure compared with their control samples, whereas molars with traditional endodontic access cavities required significantly reduced loads in order to fail. According to the numerical investigation, the type of instrumentation system was found to have an insignificant effect on the fracture resistance of the teeth under study. Von Mises stress was concentrated around the cervical region and in the larger distal roots for all numerical models. CONCLUSIONS The fracture resistance of mandibular first molars is influenced significantly by a reduction in dental hard tissue, which was found to control the level of the ultimate failure load for each tooth.