Discovery of Biomarkers for Myogenous Temporomandibular Disorders Through Salivary Metabolomic Profiling: A Pilot Study

Aims: To develop a new approach to provide insights into contributing factors to the etiology and pathogenesis of temporomandibular disorders (TMDs) through discrimination of the salivary metabolomic profiling of patients with TMDs of muscular origin (ie, local myalgia) and healthy individuals. Methods: Saliva samples from 19 patients with TMDs of muscular origin (ie, local myalgia) and 39 healthy controls were collected and identified by nuclear magnetic resonance (NMR) spectroscopy. 1H NMR spectra for all samples were acquired using a Bruker Avance-III NMR spectrometer operating at 500 MHz, and data processing was performed in TopSpin, MestreNova, SIMCA, and AMIX softwares for metabolite identification. Results: Eight key metabolites were identified between the healthy controls and patients: L-isoleucine, methylmalonic acid, isopropanolamine, dimethyl sulfone, lactic acid, 4-ethoxyphenylacetic acid, N-acetyl alanine, and D-galactose. Conclusions: The results of this study demonstrate that NMR-based metabolomics coupled with multivariate data analysis is a powerful method for the metabolomic profiling of patients with TMDs of muscular origin (ie, local myalgia).

temporomandibular disorders; salivary diagnosis; NMR analysis; metabolomics

1. Scrivani SJ, Keith DA, Kaban LB. Temporomandibular disorders. New England J Med 2008;359:2693–2705.

2. Okeson JP. Pains of muscle origin. In: Bell’s Oral and Facial Pain, ed 7. Quintessence, 2014:287–326.

3. Shrivastava M, Battaglino R, Ye L. A comprehensive review on biomarkers associated with painful temporomandibular disorders. Int J Oral Sci 2021;13:23.

4. Singh R, Kaur S. Etiology of temporomandibular disorder—A brief review: Part I. J Adv Med Dent Sci Res 2018;6:78–81.

5. Azhar F, Sani A, Baig S. Multiple etiologies of temporomandibular joint disorders—A review. Pak J Med Dent 2019;8:53–57.

6. Carlsson GE. Epidemiology and treatment need for temporomandibular disorders. J Orofac Pain 1999;13:232–237.

7. Kandasamy S, Greene CS. The evolution of temporomandibular disorders: A shift from experience to evidence. J Oral Pathol Med 2020;49:461–469.

8. Emodi-Perlman A, Eli I, Smardz J, et al. Temporomandibular disorders and bruxism outbreak as a possible factor of orofacial pain worsening during the covid-19 pandemic—Concomitant research in two countries. J Clin Med 2020;9:3250.

9. Kaufman E, Lamster IB. The diagnostic applications of saliva—A review. Crit Rev Oral Biol Med 2002;13:197–212.

10. Pfaffe T, Cooper-White J, Beyerlein P, Kostner K, Punyadeera C. Diagnostic potential of saliva: Current state and future applications. Clin Chem 2011;57:675–687.

11. Lee Y-H, Wong DT. Saliva: An emerging biofluid for early detection of diseases. Am J Dent 2009;22:241–248.

12. Lalue Sanches M, Sforça ML, Guimarães Lo Turco E, Faber J, Smith RL, Carvalho de Moraes LO. 1H-NMR-based salivary metabolomics from females with temporomandibular disorders—A pilot study. Clin Chimica Acta 2020;510:625–632.

13. Schiffman E, Ohrbach R, Truelove E, et al. Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) for clinical and research applications: Recommendations of the International RDC/TMD Consortium Network and Orofacial Pain Special Interest Group. J Oral Facial Pain Headache 2014:28:6–27.

14. Goldberg DP. A User’s Guide to the General Health Questionnaire. NFER-NELSON, 1988.

15. Billoir E, Navratil V, Blaise BJ. Sample size calculation in metabolic phenotyping studies. Brief Bioinform 2015;16:813–819.

16. Bueno CH, Pereira DD, Pattussi MP, Grossi PK, Grossi ML. Gender differences in temporomandibular disorders in adult populational studies: A systematic review and meta-analysis. J Oral Rehabil 2018;45:720–729.

17. Wheelock ÅM, Wheelock CE. Trials and tribulations of ‘omics data analysis: Assessing quality of SIMCA-based multivariate models using examples from pulmonary medicine. Mol Biosyst 2013;9:2589–2596.

18. Zhang X, Shi Y, Wang L, et al. Metabolomics for biomarker discovery in fermented black garlic and potential bioprotective responses against cardiovascular diseases. J Agric Food Chem 2019;67:12191–12198.

19. Geng C, Guo Y, Qiao Y, et al. UPLC-Q-TOF-MS profiling of the hippocampus reveals metabolite biomarkers for the impact of Dl-3-n-butylphthalide on the lipopolysaccharide- induced rat model of depression. Neuropsychiatr Dis Treat 2019;15:1939–1950.

20. Gu C, Mao X, Chen D, Yu B, Yang Q. Isoleucine plays an important role for maintaining immune function. Curr Protein Pept Sci 2019;20:644–651.

21. Kanehira H, Agariguchi A, Kato H, Yoshimine S, Inoue H. Association between stress and temporomandibular disorder. Nihon Hotetsu Shika Gakkai Zasshi 2008;52:375–380.

22. Salameh E, Alshaarani F, Hamed HA, Nassar JA. Investigation of the relationship between psychosocial stress and temporomandibular disorder in adults by measuring salivary cortisol concentration: A case-control study. J Indian Prosthodont Soc 2015;15:148–152.

23. Solís-Ortiz S, Arriaga-Avila V, Trejo-Bahena A, Guevara-Guzmán R. Deficiency in the essential amino acids l-isoleucine, l- leucine and l-histidine and clinical measures as predictors of moderate depression in elderly women: A discriminant analysis study. Nutrients 2021;13:3875.

24. Wishart DS, Guo AC, Oler E, et al. HMDB 5.0: The Human Metabolome Database for 2022. Nucleic Acids Res 2022;50:D622–D631.

25. Cömert Kılıç S. Does glucosamine, chondroitin sulfate, and methylsulfonylmethane supplementation improve the outcome of temporomandibular joint osteoarthritis management with arthrocentesis plus intraarticular hyaluronic acid injection. A ran-domized clinical trial. J Craniomaxillofac Surg 2021;49:711–718.

26. Ahn H, Kim J, Lee M-J, Kim YJ, Cho Y-W, Lee G-S. Methylsulfonylmethane inhibits NLRP3 inflammasome activa-tion. Cytokine 2015;71:223–231.

27. Butawan M, Benjamin RL, Bloomer RJ. Methylsulfonylmethane: Applications and safety of a novel dietary supplement. Nutrients 2017;9:290.

28. Schwane JA, Johnson SR, Vandenakker CB, Armstrong RB. Delayed-onset muscular soreness and plasma CPK and LDH activities after downhill running. Med Sci Sports Exerc 1983;15:51–56.

29. Gladden LB. Lactate metabolism: A new paradigm for the third millennium. J Physiol 2004;558:5–30.

30. Doshi TL, Nixdorf DR, Campbell CM, Raja SN. Biomarkers in temporomandibular disorder and trigeminal neuralgia: A conceptual framework for understanding chronic pain. Can J Pain 2020;4:1–18.

31. Shoji Y, Choo HL, Leong CO, Oo AL, Townsend G. Orofacial pain of muscular origin is not associated with herpes virus-6 infection: A pilot study. J Oral Facial Pain Headache 2014;28:346–349.

32. Milam S. Pathophysiology and epidemiology of TMJ. J Musculoskeletal Neuronal Interact 2003;3:382–390.

33. Wadhwa S, Kapila S. TMJ disorders: Future innovations in diagnostics and therapeutics. J Dent Educ 2008;72:930–947.