This study investigated the potential of salivary bacterial and protein markers for evaluating the disease status in healthy individuals or patients with gingivitis or caries. Saliva samples from caries- and gingivitis-free individuals (n = 18), patients with gingivitis (n = 17), or patients with deep caries lesions (n = 38) were collected and analyzed for 44 candidate biomarkers (cytokines, chemokines, growth factors, matrix metalloproteinases, a metallopeptidase inhibitor, proteolytic enzymes, and selected oral bacteria). The resulting data were subjected to principal component analysis and used as a training set for random forest (RF) modeling. This computational analysis revealed four biomarkers (IL-4, IL-13, IL-2-RA, and eotaxin/CCL11) to be of high importance for the correct depiction of caries in 37 of 38 patients. The RF model was then used to classify 10 subjects (five caries-/gingivitis-free and five with caries), who were followed over a period of six months. The results were compared to the clinical assessments of dental specialists, revealing a high correlation between the RF prediction and the clinical classification. Due to the superior sensitivity of the RF model, there was a divergence in the prediction of two caries and four caries-/gingivitis-free subjects. These findings suggest IL-4, IL-13, IL-2-RA, and eotaxin/CCL11 as potential salivary biomarkers for identifying noninvasive caries. Furthermore, we suggest a potential association between JAK/STAT signaling and dental caries onset and progression.

Academic Centre for Dentistry Amsterdam Department of Oral Biochemistry Free University of Amsterdam and University of Amsterdam 1081 LA Amsterdam The Netherlands

Austrian Institute of Technology Molecular Diagnostics Giefinggasse 4 1210 Wien Austria

BioVendor Laboratorní Medicína a s Research and Diagnostic Products Division Immunoassays Clinical Validation and BioVendor Analytical Testing Service Karasek 1767 1 62100 Brno Czech Republic

Clinic of Conservative and Preventive Dentistry Center of Dental Medicine University of Zurich Plattenstrasse 11 8032 Zurich Switzerland

Department of Dental Medicine Division of Oral Diseases Karolinska Institutet 141 04 Huddinge Sweden

Department of Medical Microbiology and Infectious Diseases Erasmus University Medical Centre Rotterdam 3015 GD Rotterdam The Netherlands

Hahn Schickard Georges Koehler Allee 103 79110 Freiburg Germany

Laboratory for MEMS Applications IMTEK Department of Microsystems Engineering University of Freiburg Georges Koehler Allee 103 79110 Freiburg Germany

Statistical Services Center of Dental Medicine University of Zurich Plattenstrasse 11 8032 Zurich Switzerland

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