The aim of this study was to evaluate the changes in T2 values and apparent diffusion coefficient (ADC) in the masseter muscle by clenching in healthy volunteers. 37 volunteers were enrolled in the study. We measured bite force using pressure-sensitive paper and a T2 map. The ADC map was obtained at rest, during clenching, immediately after and 5 min after clenching. The spin-echo sequence was used to calculate T2, and single-shot spin-echo echo planar imaging was used to calculate the ADC. The motion-probing gradients (MPGs) were applied separately along the posterior-to-anterior (PA), right-to-left (RL) and superior-to-inferior (SI) directions, with b values of 0, 300 and 600 s mm(-2) in each direction. ADC-PA, ADC-RL, and ADC-SI values were obtained, and we calculated the ADC-iso for the mean diffusivity. There were no significant differences between the stronger and weaker sides of bite force before, during or 5 min after clenching for T2 and ADC. The bite force had little effect on these parameters; thus, we used the average of the two sides for the following analyses. Time course analysis of ADC-iso, ADC-PA, ADC-RL and ADC-SI demonstrated a marked increase after clenching and a rapid decrease immediately after clenching, although they did not completely return to the initial values; however, the change in ADC-RL was significantly greater than those in ADC-PA or ADC-SI (P<0.001 each). The changes in T2 were similar to those of ADC, although not as marked. ADC (especially ADC-RL) was altered by contraction of the masseter muscle.