A total of 15 healthy men participated in a two-session-experiment: In one session, they performed 30 min controlled eccentric jaw exercise and the other session served as a no-exercise control. Soreness sensations at rest and during maximal biting, pressure pain thresholds (PPTs) and electromyographic (EMG) activity during maximal jaw biting were recorded before (baseline), immediately after (Post-task), and 1 day after the exercise (1-day-after). The temporal summation ratio using intra-muscular electrical stimulation of the masseter was investigated at baseline and at 1-day-after. The eccentric jaw exercise was associated with significant increases in soreness sensation and decreased PPTs at Post-task and at 1-day-after. The EMG activity and biting force did not change. The summation ratio was significantly decreased at 1-day-after in both sessions. The present findings demonstrate that eccentric jaw exercise does not induce detectable changes in temporal summation. However, the summation ratio may have clinical utility to differentiate the location of sensitization.