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. 2020 Aug 6;10(8):318.
doi: 10.3390/metabo10080318.

Metabolic Profiles of Whole, Parotid and Submandibular/Sublingual Saliva

Marco Meleti  1   2 , Eleonora Quartieri  2   3 , Rita Antonelli  1 , Margherita E Pezzi  1 , Benedetta Ghezzi  1 , Maria Vittoria Viani  1 , Giacomo Setti  1 , Emanuela Casali  2 , Elena Ferrari  2 , Tecla Ciociola  2 , Alberto Spisni  2 , Thelma A Pertinhez  2   3
Affiliations

Metabolic Profiles of Whole, Parotid and Submandibular/Sublingual Saliva

Marco Meleti et al. Metabolites. .

Abstract

The detection of salivary molecules associated with pathological and physiological alterations has encouraged the search of novel and non-invasive diagnostic biomarkers for oral health evaluation. While genomic, transcriptomic, and proteomic profiles of human saliva have been reported, its metabolic composition is a topic of research: metabolites in submandibular/sublingual saliva have never been analyzed systematically. In this study, samples of whole, parotid, and submandibular/sublingual saliva from 20 healthy donors, without dental or periodontal diseases, were examined by nuclear magnetic resonance. We identified metabolites which are differently distributed within the three saliva subtypes (54 in whole, 49 in parotid, and 36 in submandibular/sublingual saliva). Principal component analysis revealed a distinct cluster for whole saliva and a partial overlap for parotid and submandibular/sublingual metabolites. We found exclusive metabolites for each subtype: 2-hydroxy-3-methylvalerate, 3-methyl-glutarate, 3-phenylpropionate, 4-hydroxyphenylacetate, 4-hydroxyphenyllactate, galactose, and isocaproate in whole saliva; caprylate and glycolate in submandibular/sublingual saliva; arginine in parotid saliva. Salivary metabolites were classified into standard and non-proteinogenic amino acids and amines; simple carbohydrates; organic acids; bacterial-derived metabolites. The identification of a salivary gland-specific metabolic composition in healthy people provides the basis to invigorate the search for salivary biomarkers associated with oral and systemic diseases.

Keywords: metabolomics; parotid; saliva; salivary gland; submandibular/sublingual.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Experimental set-up used for parotid (A), submandibular/sublingual (B), and whole saliva (C) collection.
Figure 2
Figure 2
1H-NMR spectra of unstimulated WS, PS, and SM/SL saliva from the same participant, acquired at 25 °C. The left region of the spectra shows the vertical scale increased by a factor of 10.
Figure 3
Figure 3
PCA score plot of 1H-NMR spectra of whole saliva (blue), parotid saliva (red), and submandibular/sublingual saliva (green) samples. The contribution of the three principal components of the total variance are PC1 = 62.6%, PC2 = 6.8%, and PC3 = 5.1%.
Figure 4
Figure 4
Heatmap analysis of metabolites grouped according to the categories: (A) amino acids, (B) carbohydrates, (C) organic acids, and (D) selected prokaryotic metabolites. PS (red), SMS (green), and WS (blue) samples.
Figure 5
Figure 5
Eukaryotic (A) and prokaryotic (B) cell count distributions obtained with the three salivary types of all the subjects of the study. The significance level of the independent t-tests is set at 0.05 (* p < 0.05).
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