Temporomandibular disorders (TMDs) are associated with perturbation of proprioceptive and nociceptive function. Recent studies have shown that injection of the neurotrophic protein nerve growth factor (NGF) into the masseter muscle causes sensitization to mechanical pressure stimuli; however, it is not clear if vibration sense and jaw stretch reflexes as measures of proprioceptive function as well as glutamate-evoked pain are also altered. We tested the hypothesis that NGF-induced mechanical sensitization would be associated with changes in vibration sense and stretch reflex sensitivity as well as facilitation of glutamate-evoked pain responses. A double-blind, randomized and placebo-controlled study was conducted on 14 healthy men. In one session subjects received an injection of NGF (5 microg in 0.2 ml) into the masseter muscle and in a control session an injection of buffered isotonic saline (0.9%, 0.2 ml). Subjects assessed their pain intensity on a 0-10 cm visual analogue scale (VAS) for 15 min after the injections. Pressure pain thresholds (PPT), vibration sense and jaw stretch reflexes were recorded at baseline and 1, 2, 3 and 24 h post-injection. The sensitivity to injections of glutamate into the masseter muscle (1 M, 0.2 ml) was assessed after 24 h. ANOVAs were used to assess significant differences. NGF did not cause more pain than isotonic saline, but significantly reduced PPTs 1, 2, 3 and 24 h post-injection (P < 0.001) whereas isotonic saline had no effects on PPTs (P = 0.583). The injection of glutamate after 24 h was associated with reduced PPTs in both sessions, but the PPTs remained lower in the NGF pretreated masseter than in the control masseter (P < 0.001). Ratings of vibratory stimuli and the normalized amplitude of the jaw stretch reflex were not affected by the NGF-induced sensitization; however, after glutamate injection a significant increase in the stretch reflex was observed in the injected masseter muscle in both sessions (P = 0.002). There were no significant differences in the perceived pain intensity of the glutamate injection between the masseter muscle pretreated with NGF or control (P > 0.414), although the glutamate-evoked pain drawing areas were larger for the NGF-pretreated masseter muscle (P = 0.009). In conclusion, this study confirms that masseter muscle injection of NGF is associated with a distinct and prolonged sensitization to mechanical stimuli, but without an effect on large-diameter mechanoreceptive and the muscle spindle afferents. Additional challenge of the NGF pretreated muscle with glutamate did not indicate a conspicuous sensitization to noxious chemical stimuli. These findings are discussed in terms of the concept of "proprioceptive allodynia".