Ultrasensitive Multiplex Salivary Cytokine Profiling Reveals Distinct Inflammatory Patterns in Dental Caries

BACKGROUND Dental caries is a prevalent chronic disease characterized by complex inflammatory responses in the oral environment. This case-control pilot study evaluated the analytical performance of an ultra-sensitive electrochemiluminescence multiplex assay (MSD S-PLEX®) for salivary cytokine profiling and explored whether combined cytokine signatures can discriminate dental caries lesions (CL) from healthy (H) and non-carious gingival inflammation (GI). METHODS Unstimulated whole saliva from 79 individuals (H, CL, GI) was analyzed using the S-PLEX® assay to quantify nine salivary cytokines (IL-1β, TNF-α, IL-6, IL-4, IL-10, IL-12p70, IFN-γ, IL-17, and IL-2). Group comparisons were performed using Mann-Whitney tests, and diagnostic performance was assessed by logistic regression and the area under the receiver operating characteristic curve. RESULTS The assay showed high sensitivity and reproducibility. Cytokine profiling revealed selective elevation of certain cytokines in saliva from CL. Among the models tested, TNF-α + IL-17 provided the best discrimination between CL and H (AUC = 0.901, p  < 0.001) and between CL and H + GI (AUC = 0.795, p  = 0.0003). Inclusion of additional cytokines did not improve performance. However, discrimination between CL and GI was limited with TNF-α + IL-17, while IL-1β + IL-17 provided a modest but significant discrimination (AUC = 0.737, p  = 0.004). CONCLUSION This study provides evidence that ultrasensitive multiplex salivary cytokine profiling can reveal caries-associated inflammatory patterns. These findings highlight the translational potential of salivary cytokine panels as complementary tools for improved discrimination of caries and underline their potential for advancing precision approaches in caries research and management.