To investigate ex vivo the influence of gamma irradiation therapy and restorative material on fracture resistance, fracture mode and strain of root filled human premolars. Sixty extracted human maxillary premolar teeth were randomly divided into six groups (n = 10) determined by two study factors: (i) restorative materials: sound teeth, root filled teeth restored with composite resin, root filled teeth restored with amalgam; (ii) gamma irradiation: irradiated (subjected to 60 Gy of gamma irradiation in daily increments of 2 Gy) and nonirradiated. For the strain gauge test, two strain gauges per sample were attached on the buccal and palatal cusp surfaces (n = 5). Strain values were recorded during loading of 0-150 N. Fracture resistance (N) was assessed in a mechanical testing machine (n = 10). Strain gauge for each cusp and fracture resistance data were analysed by two-way anova (3 × 2) followed by the Tukey's honestly significant difference test (α = 0.05). The failure mode was evaluated using an optical stereomicroscope and classified according to the location of the failure. Gamma radiation therapy significantly reduced the fracture resistance of intact teeth. The strain was higher for teeth restored with amalgam than for those restored with composite resin. The teeth restored with composite resin had similar strain values to sound teeth. Nonirradiated teeth had more restorable failures than irradiated teeth. Gamma irradiation significantly reduced fracture resistance and increased cusp strain. The use of composite resin resulted in better biomechanical behaviour than amalgam for restoring root filled teeth whether or not they were submitted to radiotherapy.