In this experimental study, we aimed to determine whether guided music listening (GML) - a music intervention based on models of mood mediation and attention modulation - modulates masticatory muscle activity and awake bruxism in subjects with chronic painful muscular temporomandibular disorders (TMD myalgia, mTMD), a condition causing a significant burden to patients, their families, and healthcare systems. Awake bruxism - a stress behavior characterized by clenching of the teeth - is a strong contributor to chronic mTMD. GML modulates psychological stress and motor responses and could thus reduce muscle activity in chronic musculoskeletal conditions, including mTMD. We recorded the electromyographic (EMG) activity in the right masseter of 14 women with chronic (>6months) mTMD (median [IQR] = 39.5.3 [24.3] years) and 15 pain-free women (median [IQR] = 30.0 [3.5] years) during a GML session, including 3 music (stressful, relaxing, and participants' favorite music) and a no-music (pink noise) control blocks, each lasting 15minutes. We measured the motor effort of the right masseter relative to the participants' maximum voluntary contraction (MVC), the muscular effort to maintain mandibular posture (EMGposture ), and to produce spontaneous awake bruxism episodes (EMGbruxism ), and the duration and frequency of spontaneous awake bruxism episodes. We tested between-group and within-group (between blocks) differences, as well as the effect of the interaction group by experimental block on these outcome measures. In both groups, EMGposture was significantly affected by the interaction group by experimental block (P<.001). Compared to pink noise [mean (95% CI); mTMD: 2.2 (1.6-2.8) %MVC; Controls: 1.1 (0.5-1.7) %MVC], EMGposture increased during the stressful music block [contrast estimate (95% CI); mTMD: +0.8 (0.7-0.8) %MVC; Controls: +0.3 (0.3-0.4) %MVC; both P<.001], and decreased during the relaxing [mTMD: -0.4 (-0.5 to -0.4) %MVC; Controls: -0.3 (-0.4 to -0.3) %MVC; both P<.001] and favorite [mTMD: -0.5 (-0.6 to -0.5) %MVC; Controls: -0.5 (-0.5 to -0.4) %MVC; both P<.001] music blocks. EMGposture was greater in mTMD individuals than controls during the favorite music [contrast estimate (95% CI): +1.1 (0.2-1.9) %MVC; P=.019] and the pink noise [+1.1 (0.2-2.0) %MVC; P=.014] blocks. EMGbruxism was significantly affected by the interaction group by experimental block (P<.001). In mTMD participants, compared to the pink noise block [mean (95% CI); 23.8 (16.0-31.6) %MVC], EMGbruxism increased during the stressful music block [contrast estimate (95% CI); +10.2 (8.6-11.8) %MVC], and decreased during the relaxing [-6.2 (-8.1 to -4.3) %MVC; P<.001] and favorite [-10.2 (-12.2 to -9.1) %MVC; P<.001] music blocks. These effects were not observed in the control group [mean (95% CI); pink noise: 19.3 (10.9-27.6); stressful: 21.2 (12.9-29.4) %MVC; relaxing: 21.6 (13.3-29.9) %MVC; favorite: 24.2 (15.8-32.7) %MVC; all P >.05]. EMGbruxism was significantly greater in mTMD participants than controls during the stressful music block [contrast estimate (95% CI): +12.9 (1.6-24.2) %MVC; P=.026). GML did not affect the duration or the frequency of awake bruxism in either group (median [IQR], mTMD: 23.5 [96.7] s, range 1-1300seconds; Controls: 5.5 [22.5], range 0-246seconds; P=.108). The frequency of awake bruxism episodes was greater in the mTMD group compared to controls only during the pink noise block (median [IQR], mTMD: 5 [15.3] episodes, range 0-62 episodes; Controls: 1 [3] episode, range 0-27 episodes; P=.046). No significant between-group differences were found in either the overall time spent engaging in awake bruxism (median [IQR], mTMD: 23.5 [96.7] s, range 1-1300seconds; Controls: 5.5 [22.5], range 0-246seconds; P=.108), or during each block (all P >.05). In subjects with chronic mTMD, relaxing music and the individual's favorite music decreased the muscular effort during spontaneous awake bruxism episodes by 26% and 44% (relative changes), respectively. In contrast, stressful music increases it by about 43%. Because of its positive effects on awake bruxism, GML with selected music could be a promising and non-invasive component of a multimodal approach for the management of chronic mTMD.