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Background and aims: Muscle related temporomandibular disorders (myogenous TMD), one of the most common orofacial pain conditions, is characterized by facial pain and often accompanied by jaw movement limitations. Although the underlying biological mechanisms are still unclear, a cluster of proteins and peptides is assumed to be involved in the pathophysiology. These proteins and peptides may be measured in a simple non-invasive saliva sample. This work investigated whether saliva can be used to sample algogenic substances that can serve as molecular biomarkers for TMD myalgia. Methods: Saliva and blood samples were collected from healthy individuals (n=69) and patients diagnosed with TMD myalgia (n=39) according to the Diagnostic Criteria for TMD. Unstimulated and stimulated whole, parotid, and sublingual saliva were analysed. The protein profiles were investigated using two-dimensional gel electrophoresis followed by identification with liquid chromatography tandem mass spectrometry. Levels of nerve growth factor (NGF), calcitonin gene-related peptide (CGRP), and brain derived neuro-tropic factor (BDNF) were determined using western blotting based technology and multiplex electrochemiluminescence assay panel. Glutamate, serotonin, and substance p (SP) were determined using commercially available methods. Results: Different saliva collection approaches resulted in significant differences in the protein profile as well as in the expression of NGF, BDNF, CGRP, SP, and glutamate. Stimulated whole saliva showed least variability in protein concentration (35%) and was correlated to plasma levels of glutamate. Unlike SP and glutamate, NGF and BDNF expressed a rhythmic variation in salivary expression with higher levels in the morning (p<0.05). Patients with a diagnosis of TMD myalgia had significantly higher levels of salivary glutamate but lower salivary NGF and BDNF compared to controls; in addition, the lower NGF and BDNF levels correlated to psychological dysfunction. The quantitative proteomics data revealed 20 proteins that were significantly altered in patients compared to controls. The identified proteins are involved in metabolic processes, immune response, and stress response. Dissimilarities in protein profile and clinical variables were observed between TMD myalgia and myofascial pain. Conclusions: The work highlights the importance of consistency in saliva collection approaches, including the timing of the collection. It displayed significant changes in pain specific mediators and protein profile in TMD myalgia and furthermore dissimilarities between subclasses indicating different pathophysiology. After extensive validation, potential salivary biomarkers can be combined with clinical features to better understand and diagnose TMD myalgia. [ABSTRACT FROM AUTHOR]

Background: The clinical care of chronic pain requires personalised understanding of the mechanisms involved. Temporomandibular disorders (TMD) are the most common chronic orofacial pain conditions, and oxidative stress has been proposed to be implicated in their pathophysiology, especially in arthrogenous TMD. However, few studies have explored oxidative stress in myogenous TMD (TMDM). Objective: The aims of this study were to compare the salivary oxidative stress profiles between individuals with TMDM and healthy controls, and to explore associations of these markers with clinical characteristics. Methodology: Saliva samples were collected from 39 individuals with TMDM and 37 age and sex‐matched healthy volunteers. Psychological stress levels and clinical characteristics were assessed in all participants. The samples were analysed for total oxidant status (TOS), total antioxidative capacity (TAC) and superoxide dismutase activity (SODa). Comparisons between groups were performed using parametric and non‐parametric tests depending on data distribution. Results: Psychological stress was higher in TMDM compared to controls (P <.001). TAC levels were significantly higher (P <.05) whereas TOS levels were significantly lower (P <.05) in TMDM compared to controls. There were no differences in SODa levels between groups and no correlations were found between clinical characteristics and oxidative stress markers. Conclusion: Individuals with TMDM showed higher levels of antioxidative markers, but lower levels of oxidative markers. These results can be explained in part by chronicity and adaptation to the disease and other factors, such as psychological stress. Longitudinal studies must be conducted to clarify the role of oxidative stress in TMDM. [ABSTRACT FROM AUTHOR]