What question did this study set out to answer?

This study aims to assess the feasibility of using salivary DNA methylation as biomarkers for diagnosing and stratifying Temporomandibular Disorders (TMD).

May 9, 2026Open Access

38 Salivary DNA methylation signatures as potential biomarkers for chronic painful temporomandibular disorders: A pilot study

Key Points

This study aims to assess the feasibility of using salivary DNA methylation as biomarkers for diagnosing and stratifying Temporomandibular Disorders (TMD).
Cross-sectional pilot study enrolling 13 TMD patients and 4 controls from a pain clinic.
Saliva samples analyzed with Illumina EPIC BeadChip arrays to explore ~850,000 CpG sites.
Bioinformatics used for principal component analysis and identification of differentially methylated sites.
Principal component analysis indicated a clear clustering between TMD patients and controls.
Methylation profile scores successfully differentiated between high-impact and low-impact TMD subgroups.
High-impact TMD patients had significantly higher pain intensity and psychological scores compared to low-impact patients.

Abstract

Objectives/Goals: To evaluate the feasibility of developing salivary DNA methylation-based biomarkers for objective diagnosis and stratification of Temporomandibular Disorders (TMD), addressing the critical need for quantitative diagnostic tools to complement current subjective assessments and clinical examinations. Methods/Study Population: This cross-sectional pilot study enrolled 13 TMD patients and 4 pain-free controls from the University of Minnesota TMD Orofacial Pain Clinic. Participants underwent comprehensive DC/TMD diagnostic evaluation and completed validated instruments measuring pain intensity (VAS), psychological status (PHQ-9, PCS), and functional impairment (GCPS). Saliva samples were analyzed using Illumina EPIC BeadChip arrays interrogating ~850,000 CpG sites. Bioinformatic analyses included principal component analysis, identification of differentially methylated sites, and development of composite Methylation Profile Scores for patient stratification Results/Anticipated Results: Principal component analysis showed clear clustering separation between TMD patients and controls. Methylation profile scores demonstrated clear stratification between controls, high-impact, and low-impact TMD subgroups. High-impact TMD participants (n=6) showed significantly elevated clinical severity scores compared to low-impact TMD (n=7): pain intensity (52.0±11.3 vs 25.0±13.8), depression (20.5±7.3 vs 8.4±4.9), and catastrophizing (12.6±8.1 vs 6.4±6.5). Promoter-level analysis identified candidate genes associated with TMD severity phenotypes, supporting the potential for epigenetic biomarkers to enable objective patient stratification and guide precision treatment selection. Discussion/Significance of Impact: Epigenetic biomarkers capturing TMD heterogeneity could transform clinical practice by enabling early-risk stratification, preventing progression to high-impact chronic pain through personalized interventions, and reducing the substantial healthcare burden of this prevalent orofacial pain condition.

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