BioMed Research International

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Research on Saliva Secretion and Composition

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Volume 2018 |Article ID 3921247 |

Zeineb Zian, Joaira Bakkach, Amina Barakat, Naima Ghailani Nourouti, Mohcine Bennani Mechita, "Salivary Biomarkers in Systemic Sclerosis Disease", BioMed Research International, vol. 2018, Article ID 3921247, 7 pages, 2018.

Salivary Biomarkers in Systemic Sclerosis Disease

Academic Editor: Fernando C. e Silva
Received19 Oct 2017
Revised14 Jan 2018
Accepted11 Feb 2018
Published12 Mar 2018


Scleroderma or systemic sclerosis (SSc) is frequently detected at an advanced stage due to diagnosis difficulties. Salivary biomarkers, if existing, could be used for predictive diagnosis of this disease. Human saliva contains a large number of proteins that can be used for diagnosis and are of great potential in clinical research. The use of proteomic analysis to characterize whole saliva (WS) in SSc has gained an increasing attention in the last years and the identification of salivary proteins specific for SSc could lead to early diagnosis or new therapeutic targets. This review will present an overview about the use of WS in SSc studies. The proteomic technologies currently used for global identification of salivary proteins in SSc, as well as the advantages and limitations for the use of WS as a diagnostic tool, will be presented.

1. Introduction

Scleroderma or systemic sclerosis (SSc) is a rare systemic autoimmune disease affecting the connective tissue and characterized by involvement of the skin, blood vessels, and visceral organs. It is associated with dysfunction of the immune cells, fibroblasts, and endothelial cells. The etiology of SSc remains unknown [1]. SSc affects preferentially women, more often during and after their childbearing age [2]. Sex ratio varies in published series from 3 to 9 women for a man [3].

There are two main clinical forms of SSc that differ primarily in their degree of skin involvement: limited cutaneous scleroderma (lcSSc) and diffuse cutaneous scleroderma (dcSSc), which are associated with different clinical complications [4]. Oral manifestations are frequent in SSc [5], and the majority of oral clinical features start with tongue rigidity and facial skin hardening [6]. On the other hand, it was shown that SSc affects salivary glands [7], and these latter can also be subject to fibrosis in SSc patients [8].

The medications and fibrotic changes in salivary glands of patients with SSc can contribute to reduced salivary flow in these patients [9]. This diminishing of saliva production in SSc patients is mostly related to concomitant Sjögren’s syndrome [10]. Although a little knowledge about salivary gland involvement in SSc has been reported in the literature, it was demonstrated, in a previous study, that salivary gland changes (increased expression of E-selectin and TNF-α and infiltration by mast cells) are detectable in the early stages of the disease, before the onset of skin changes and when the criteria for a diagnosis of SSc are absent [11].

The identification of salivary protein profiles could lead to early diagnosis or new therapeutic targets of SSc [12]. Furthermore, the presence of the biomarker may correlate with different clinical symptoms of the disease due to its absence in healthy subjects [13]. In the last years and with technological and analytical development, saliva has attracted an increased interest for use in diseases diagnosis and treatment. To date, there have been few reports aimed to use the WS in SSc research [7, 1214]. Giusti et al. [14] performed, for the first time, a study in an attempt to characterize the WS protein profiles of patients with SSc using a proteomic research approach.

In this review we will give an overview of the use of WS in SSc research. The proteomic technologies currently used for global identification of salivary proteins in SSc, as well as the advantages and limitations for the use of WS in the disease, will be presented.

2. Whole Saliva

A number of proteomics researches contributed to the clarification and knowledge of the salivary proteome, and more than 2000 proteins and peptides have been found in WS and individual salivary glands [15]. Saliva is a mucoserous exocrine fluid produced by three major salivary glands (parotid, submandibular (SM), and sublingual (SL)) and other minor glands located under the oral mucosa [16]. Besides, WS comes also from local and systemic sources [17]. It is a combination of the secretions from the major and minor salivary glands, oral mucosa transudate, the gingival fold, desquamated epithelial and blood cells, nasal secretions, viruses, fungi, bacteria, and food debris [18]. WS contains hormones, immunoglobulins, proteins, enzymes, and mucosal glycoproteins [18]. It also contains a number of antimicrobial proteins which play an important role in reducing oral infections [19]. Its role in protecting oral structures has been well reported. Although saliva includes blood derived products, differently to serum, this oral fluid contains many locally secreted proteins which may be specific markers for some local diseases [16]. The presence in WS of many molecules that are circulating in the blood presents several advantages for disease diagnosis and prognosis as the collection of this fluid is noninvasive, safe, and easy; relatively low amounts of sample are needed and storage and transportation are not complicated [20, 21]. That is why, nowadays, WS is used as a diagnostic tool in clinical diagnosis, monitoring disease progression and management of patients [22, 23].

Passive drool saliva collection method is considered as the gold standard, but there are other saliva collection devices such as Salivettes [24, 25] which present a less viscosity and allow an easy handling as well as a better sample processing, particularly in some special cases such Xerostomia [26]. Unlike blood, due to its noncoagulating nature, saliva is easier to handle for diagnostic analysis procedures. The noninvasive collection procedures for saliva contribute to the procurement of repeated samples to follow the patients over time. Various collection and storage protocols of WS were described in published studies [7, 1214, 2736]. Due to the presence of circadian rhythms in WS flow rate and composition, WS collection should be made under standard conditions [37]. The variations in collection and/or storage procedures can change the salivary proteomic profiles after collection and therefore alter the biomarkers content and their detection [21], from where the need for adopting standardized procedures in saliva analysis. In fact, in order to avoid protein degradation, some authors have added 0.2% trifluoroacetic acid (TFA) to saliva sample [38]. Others have used a protease inhibitor cocktail and have stored the saliva at −20°C [39]. Furthermore, minimizing the time between collection and analysis of the sample has been proposed by some groups research [1214, 36, 40]. Thus, it has been indicated that instead of using chemical inhibitors of proteolysis, collection of saliva into ice cold tubes could minimize proteolysis, as well as storage of the samples at −80°C rather than −20°C [41], in particular for storage at longer durations [42].

3. Applications of Whole Saliva for Systemic Sclerosis Study

In the recent years, proteomic approaches started to be used in the study of WS from patients with SSc. Knaś et al. [7] study was the first to describe the alteration on the salivary glands function in both subsets of SSc.

Among the different proteomic approaches, saliva has been studied using several techniques, either separately, or, more often, in combination such as one-dimensional polyacrylamide gel electrophoresis (1D-PAGE), two-dimensional polyacrylamide gel electrophoresis/mass spectrometry (2-DE/MS), capillary electrophoresis (CE), 2D-liquid chromatography/mass spectrometry (2D-LC/MS), nuclear magnetic resonance (NMR), matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS), surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF/MS), Western blot, electrospray ionization (ESI), immunoassays (radio-immune assays, immunoradiometric assays, enzyme immuno-assays, and enzyme-linked immunosorbent assays) and lectin binding assays on PAGE gels [15, 22, 4354]. The choice for the technique is depending on the objectives of the study and on the salivary proteins of interest. It has been reported that 2-DE combined with mass spectrometry has been widely used to study salivary proteins [14, 15, 49]. However, due to its limitations it is not the most important tool used in this field and does not allow the study of the complete proteome. In fact, other techniques have been shown to be more performant, including a variety of chromatographic combinations that has successfully characterized more than 3000 different components in saliva [38, 5557]. Surface chromatography combined with MALDI-TOF/MS has allowed rapid and high-throughput detection of important proteins and peptides [58]. From our knowledge, only four studies focused on WS in SSc have been published, and only one study [14] has identified the salivary biomarkers in these patients. In these proteins separation was achieved by using 2-DE technique with subsequent protein identification being made by MALDI-TOF-MS (Table 1).

StudySaliva samplePatients/controlsAnalytical methodsFindings

Giusti et al. [14]UWS15/152-DE, MALDI-TOF/MSPresence of 9 proteins only in SSc (calgranulin A, calgranulin B, psoriasin, Arp2/3 complex, ß2-microglobulin, TPI, GAPDH, cyclophilin A, and cystatin B).

Baldini et al. [13]UWS44/80SDS/PAGE, Western blotSignificant association of psoriasin with pulmonary involvement in dcSSc.

Knaś et al. [7]UWS/SWS97/55ELISA, Spectrophotometrically(i) In UWS of dcSSc and lcSSc:
 (1) Salivary flow, the output of total protein, and peroxidase activity were significantly lower.
 (2) sIgA and lactoferrin were significantly higher.
(ii) In SWS:
 (1) In lcSSc, the total lysozyme and peroxidase activity were significantly higher.
 (2) In dcSSc, the salivary flow was significantly lower and the total sIgA and peroxidase activity were significantly higher.

Giusti et al. [12]UWS134/74ELISASignificant correlation between salivary psoriasin and DLCO in SSc.

dcSSc, diffuse cutaneous systemic sclerosis; DLCO, diffusion capacity of carbon monoxide; ELISA, enzyme-linked immunosorbent assay; lcSSc, limited cutaneous systemic sclerosis; MALDI-TOF, matrix-assisted laser desorption ionization; MS, mass spectroscopy; SDS/PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; SSc, systemic sclerosis; SWS, stimulated whole saliva; UWS, unstimulated whole saliva; 2-DE, two-dimensional gel electrophoresis.

4. Salivary Proteins in Systemic Sclerosis

To date, there is a lack of early diagnostic markers, and the time between the diagnosis and symptom onset can be translated by years. Identification of the salivary proteins biomarkers involved in SSc may contribute to the early detection of the disease. The specific salivary markers identified so far were reported by Giusti et al. [14] in a study including 15 patients with dcSSc, in which they compared the differences between WS of SSc patients and control subjects. Indeed, it was reported that both previously identified and newly identified proteins occurred in WS of SSc patients but did not match with healthy subjects. Some of these proteins, such as keratin 6L, psoriasin, TPI, and Arp2/3 complex, might play a pathological role in SSc, suggesting that some of them may be considered as new therapeutic targets or diagnostic markers for SSc. It was found also that, except Keratin 6L, the expression of most of the called typical salivary proteins like α-amylase, prolactin-inducible protein precursor, albumin, or cystatins remained unchanged between control subjects and patients. In contrast, the same research team showed that the expression of these normal proteins was altered in Sjögren’s syndrome patients compared to the controls, with decreased levels of some salivary proteins [36] (Table 2).


Calgranulin AP05109Present in chronic inflammation and in epithelial cells constitutively or induced during dermatoses. Involved in the metabolism of arachidonic acid in human neutrophils. Seem to have a major role in inflammatory and immunological responses. May interact with components of the intermediate filaments in monocytes and epithelial cells. May play a role in inhibiting the matrix metalloproteinases activity.

Calgranulin BP06702Present in acute and in chronic inflammation. Stimulate neutrophil adhesion. Involved in the metabolism of arachidonic acid in human neutrophils. May play a role in inhibiting the protein kinases and the matrix metalloproteinases activity. May interact with components of the intermediate filaments in monocytes and epithelial cells.

Cystatin BP04080Proteinase inhibiting properties. Tightly binding reversible inhibitor of cathepsins L, H, and B.

PsoriasinP31151Present in fetal ear, skin, and tongue and human cell lines. Highly expressed in psoriasis and in other inflammatory skin diseases. Seem to participate in tumor progression. Also highly expressed in the urine of patients with bladder squamous cell carcinoma.

ß2-MicroglobulinQ6IAT8Component of the MHC class I molecules.

Cyclophilin AP62937Involved in expression, folding, and degradation of proteins. Catalyze the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.

P04406Glycolytic enzymes present in the cytoplasm. Play a role in degradation of carbohydrate and glycolysis.

Triose phosphate
P60174A highly conserved glycolytic enzyme. Mediated by glycolysis in red blood cells and in brain cells. Biosynthesis of carbohydrate and gluconeogenesis.

Actin-related protein 2/3 complex subunit 2O15144Strong candidate for the control of actin polymerization in chemotaxis.

Among those proteins, three that belong to the S100 calcium- and zinc-binding protein family related to inflammation have been identified: calgranulin A (S100A8), calgranulin B (S100A9), and psoriasin (S100A7) [12, 14, 45, 59]. S100A8 and S100A9 are mainly localized in the cytosol of neutrophils and are involved in the metabolism of arachidonic acid in human neutrophils [43, 60]. Some findings suggest that high concentrations of S100A8 and S100A9 might play a role in inhibiting the matrix metalloproteinases activity by the sequestration of zinc [59, 61]. This inhibition or reduced activity of MMP plays a crucial role in reducing extracellular matrix degradation in SSc individuals and leads to extensive fibrosis of this disease. Regarding psoriasin (S100A7), it was firstly identified by Madsen et al. [62], as a protein expressed in epithelial cells of the psoriatic skin. Increase of psoriasin expression has been also observed in WS of patients with dcSSc [14]. Although the biological effect of psoriasin in SSc remains unknown, a significant association of this protein and pulmonary involvement of dcSSc has been demonstrated [13]. Arp2/3 complex has been newly identified in WS [14]. This complex plays a role in the regulation of actin polymerization in cells, and it is necessary for neutrophil chemotaxis and phagocytosis [63]. The ß2-microglobulin is a component of MHC class I molecules which may play a role in the immune dysregulation of SSc [14]. Triose phosphate isomerase (TPI) and glyceraldehyde-3 P-dehydrogenase (GAPDH) 190 are glycolytic enzymes present in cytoplasm that may act as autoantigens in SSc and also in other autoimmune diseases such as systemic lupus erythematosus [64]. Regarding cyclophilin A, its contribution to the pathogenesis of immune-mediated endothelial activation and dysfunction was suggested by Kim et al. [65]. It is involved in the expression, folding, and degradation of proteins and catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides [66]. Cystatin B is an intracellular thiol proteinase inhibiting [14, 44, 67]. However, its role in SSc has not been reported so far.

5. Advantages and Limitations

In the recent years and with the advances in proteomic technologies, salivary research emerged as an important area for the diagnosis of several local and systemic diseases. As mentioned above, saliva showed several advantages for systemic diseases research as well as for SSc including mainly safety and easy collection using simple, inexpensive, and noninvasive methods. The presence of several serum components in saliva has great benefits for research of new biomarkers. Moreover, due to its noncoagulating nature, saliva is easier to handle in diagnostic analysis procedures [15, 20, 22, 23]. Our knowledge about specific advantages and limitations of the use of this tool with diagnosis purposes in SSc is still limited. We mention in this review a raised concern for the use of WS in SSc patients with Sjogren’s syndrome. These latter were shown to have generally a reduced salivary flow rate that could limit the collection and use of WS as research material for this group [7], but this needs to be more investigated. The most known advantages and limitations of WS that are likely to be extrapolated for SSc are presented in Table 3.


(i) Readily accessible and informative biofluid.(i) Many informative molecules in lower amounts of saliva.
(ii) Easy, safe, inexpensive, and noninvasive diagnostic approach.(ii) Centrifugation may also remove other proteins.
(iii) Noncoagulating nature.(iii) Presence of several proteases degrading protein biomarkers.
(iv) More sensitive and more specific markers for oral diseases.(iv) Difficult to have saliva completely free of stimulation which influences the results.
(v) Simple collection and minimal equipment required.(v) Possibility of assaying proteins only after recent Exposure.
(vi) Storage and transportation at low cost.(vi) Difficult detection with low concentrations of proteins of interest in saliva.
(vii) Less amounts of sample
(viii) Contains serum constituents

6. Conclusion

In conclusion, salivary biomarkers study is becoming an important part of the diagnosis of several diseases. Identification of salivary proteins in SSc is a promising finding that paved the way to new diagnostic biomarkers for this pathology, but this needs to be more investigated as there are so far only few studies published in this regard.

Several approaches (SELDI-TOF/MS, HPLC, and other affinity chromatography techniques) hold promise for salivary proteomic analysis to discover and validate new biomarkers or therapeutic targets.

Deeper and more comprehensive studies are required to elucidate the functional significance of these proteins during the SSc and to improve diagnosis as well as treatment. In addition, larger populations are needed to validate and generalize these results in future studies of salivary proteomic. On the other hand, research to the salivary proteome in SSc patients from different regions of the world is required due to the variability in genetic and environmental factors of each subject.

Conflicts of Interest

The authors declare that they have no conflicts of interest.


The authors would like to thank Professor Fernando Capela e Silva and Dr. Elsa Lamy, from ICAAM–Institute of Mediterranean Agricultural and Environmental Sciences, University of Évora, Portugal, for their valuable contribution in reviewing the manuscript.


  1. J. Martin and C. Fonseca, “The genetics of scleroderma,” Current Rheumatology Reports, vol. 13, no. 1, pp. 13–20, 2011. View at: Publisher Site | Google Scholar
  2. C. E. Weckerle and T. B. Niewold, “The unexplained female predominance of systemic lupus erythematosus: Clues from genetic and cytokine studies,” Clinical Reviews in Allergy & Immunology, vol. 40, no. 1, pp. 42–49, 2011. View at: Publisher Site | Google Scholar
  3. B. Admou and et al., “Faible prévalence des anticorps anti-centromère dans la sclérodermie au Maroc (à propos de 272 cas),” Annales de Biologie Clinique, vol. 65, no. 3, pp. 291–297, 2007. View at: Google Scholar
  4. F. D. Carmona, J. Martin, L. Beretta et al., “The Systemic Lupus Erythematosus IRF5 Risk Haplotype Is Associated with Systemic Sclerosis,” PLoS ONE, vol. 8, no. 1, p. e54419, 2013. View at: Publisher Site | Google Scholar
  5. J. B. Albilia, D. K. Lam, N. Blanas, C. M. L. Clokie, and G. K. B. Sándor, “Small mouths⋯big problems? A review of scleroderma and its oral health implications,” Journal of the Canadian Dental Association, vol. 73, no. 9, pp. 831–836, 2007. View at: Google Scholar
  6. C. Cazal, A. P. V. Sobral, R. F. N. Neves, F. W. V. Freire-Filho, A. B. Cardoso, and M. M. F. da Silveira, “Oral complaints in progressive systemic sclerosis: two cases report,” Medicina Oral Patologia Oral y Cirugia Bucal, vol. 13, no. 2, p. 114, 2008. View at: Google Scholar
  7. M. Knaś, A. Zalewska, N. Waszkiewicz et al., “Salivary: flow and proteins of the innate and adaptive immunity in the limited and diffused systemic sclerosis,” Journal of Oral Pathology & Medicine, vol. 43, no. 7, pp. 521–529, 2014. View at: Publisher Site | Google Scholar
  8. G. I. Mason, J. Hamburger, and J. B. Matthews, “Mast cells, extracellular matrix components, TGFβ isoforms and TGFβ receptor expression in labial salivary glands in systemic sclerosis,” Annals of the Rheumatic Diseases, vol. 59, no. 3, pp. 183–189, 2000. View at: Publisher Site | Google Scholar
  9. S. R. Porter, C. Scully, and A. M. Hegarty, “An update of the etiology and management of xerostomia,” Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, vol. 97, no. 1, pp. 28–46, 2004. View at: Publisher Site | Google Scholar
  10. M. Baron, M. Hudson, S. Tatibouet et al., “Relationship between disease characteristics and orofacial manifestations in systemic sclerosis: Canadian systemic sclerosis oral health study III,” Arthritis Care & Research, vol. 67, no. 5, pp. 681–690, 2015. View at: Publisher Site | Google Scholar
  11. M. Hebbar, J.-M. Gillot, E. Hachulla et al., “Early expression of E-selectin, tumor necrosis factor α, and mast cell infiltration in the salivary glands of patients with systemic sclerosis,” Arthritis & Rheumatology, vol. 39, no. 7, pp. 1161–1165, 1996. View at: Publisher Site | Google Scholar
  12. L. Giusti, F. Sernissi, E. Donadio et al., “Salivary psoriasin (S100A7) correlates with diffusion capacity of carbon monoxide in a large cohort of systemic sclerosis patients,” Journal of Translational Medicine, vol. 14, no. 1, article no. 262, 2016. View at: Publisher Site | Google Scholar
  13. C. Baldini and et al., “Association of psoriasin (S100A7) with clinical manifestations of systemic sclerosis: is its presence in whole saliva a potential predictor of pulmonary involvement?” The Journal of Rheumatology, vol. 35, no. 9, pp. 1820–1824, 2008. View at: Google Scholar
  14. L. Giusti and et al., “Specific proteins identified in whole saliva from patients with diffuse systemic sclerosis,” The Journal of Rheumatology, vol. 34, no. 10, pp. 2063–2069, 2007. View at: Google Scholar
  15. E. Lamy, A. R. Costa, C. M. Antunes, R. Vitorino, and F. Amado, “Protein Electrophoresis in Saliva Study,” in Electrophoresis, K. Ghowsi, Ed., InTech, 2012. View at: Google Scholar
  16. S. Hu, J. A. Loo, and D. T. Wong, “Human saliva proteome analysis and disease biomarker discovery,” Expert Review of Proteomics, vol. 4, no. 4, pp. 531–538, 2007. View at: Publisher Site | Google Scholar
  17. S. Podzimek, L. Vondrackova, J. Duskova, T. Janatova, and Z. Broukal, “Salivary Markers for Periodontal and General Diseases,” Disease Markers, vol. 2016, Article ID 9179632, 2016. View at: Publisher Site | Google Scholar
  18. T. Pfaffe, J. Cooper-White, P. Beyerlein, K. Kostner, and C. Punyadeera, “Diagnostic potential of saliva: current state and future applications,” Clinical Chemistry, vol. 57, no. 5, pp. 675–687, 2011. View at: Publisher Site | Google Scholar
  19. R. M. Silver and C. P. Denton, Case Studies in Systemic Sclerosis, Springer London, London, UK, 2011. View at: Publisher Site
  20. P. V. Rao, A. P. Reddy, X. Lu et al., “Proteomic identification of salivary biomarkers of type-2 diabetes,” Journal of Proteome Research, vol. 8, no. 1, pp. 239–245, 2009. View at: Publisher Site | Google Scholar
  21. E. Lamy and M. Mau, “Saliva proteomics as an emerging, non-invasive tool to study livestock physiology, nutrition and diseases,” Journal of Proteomics, vol. 75, no. 14, pp. 4251–4258, 2012. View at: Publisher Site | Google Scholar
  22. S. A. Kawas, Z. H. A. Rahim, and D. B. Ferguson, “Potential uses of human salivary protein and peptide analysis in the diagnosis of disease,” Archives of Oral Biolog, vol. 57, no. 1, pp. 1–9, 2012. View at: Publisher Site | Google Scholar
  23. A. Zhang, H. Sun, P. Wang, and X. Wang, “Salivary proteomics in biomedical research,” Clinica Chimica Acta, vol. 415, pp. 261–265, 2013. View at: Publisher Site | Google Scholar
  24. M. Gröschl, H. Köhler, H.-G. Topf, T. Rupprecht, and M. Rauh, “Evaluation of saliva collection devices for the analysis of steroids, peptides and therapeutic drugs,” Journal of Pharmaceutical and Biomedical Analysis, vol. 47, no. 3, pp. 478–486, 2008. View at: Publisher Site | Google Scholar
  25. P.-J. Lamey and A. Nolan, “The recovery of human saliva using the Salivette system,” European Journal of Clinical Chemistry and Clinical Biochemistry, vol. 32, no. 9, pp. 727-728, 1994. View at: Google Scholar
  26. L. Henderson, M. Muir, P. R. Mills et al., “Oral health of patients with hepatitis c virus infection: A pilot study,” Oral Diseases, vol. 7, no. 5, pp. 271–275, 2001. View at: Publisher Site | Google Scholar
  27. P. Dowling, R. Wormald, P. Meleady, M. Henry, A. Curran, and M. Clynes, “Analysis of the saliva proteome from patients with head and neck squamous cell carcinoma reveals differences in abundance levels of proteins associated with tumour progression and metastasis,” Journal of Proteomics, vol. 71, no. 2, pp. 168–175, 2008. View at: Publisher Site | Google Scholar
  28. Y. Fleissig, O. Deutsch, E. Reichenberg et al., “Different proteomic protein patterns in saliva of Sjögren's syndrome patients,” Oral Diseases, vol. 15, no. 1, pp. 61–68, 2009. View at: Publisher Site | Google Scholar
  29. G. Peluso, M. De Santis, R. Inzitari et al., “Proteomic study of salivary peptides and proteins in patients with Sjögren's syndrome before and after pilocarpine treatment,” Arthritis & Rheumatology, vol. 56, no. 7, pp. 2216–2222, 2007. View at: Publisher Site | Google Scholar
  30. Y. Wu, R. Shu, L.-J. Luo, L.-H. Ge, and Y.-F. Xie, “Initial comparison of proteomic profiles of whole unstimulated saliva obtained from generalized aggressive periodontitis patients and healthy control subjects,” Journal of Periodontal Research, vol. 44, no. 5, pp. 636–644, 2009. View at: Publisher Site | Google Scholar
  31. L.-L. Yang, X.-Q. Liu, W. Liu, B. Cheng, and M.-T. Li, “Comparative analysis of whole saliva proteomes for the screening of biomarkers for oral lichen planus,” Inflammation Research, vol. 55, no. 10, pp. 405–407, 2006. View at: Publisher Site | Google Scholar
  32. S. Hu, M. Arellano, P. Boontheung et al., “Salivary proteomics for oral cancer biomarker discovery,” Clinical Cancer Research, vol. 14, no. 19, pp. 6246–6252, 2008. View at: Publisher Site | Google Scholar
  33. S. Hu, J. Wang, J. Meijer et al., “Salivary proteomic and genomic biomarkers for primary Sjögren's syndrome,” Arthritis & Rheumatology, vol. 56, no. 11, pp. 3588–3600, 2007. View at: Publisher Site | Google Scholar
  34. S. Hu and et al., “Discovery of oral fluid biomarkers for human oral cancer by mass spectrometry,” Cancer Genomics-Proteomics, vol. 4, no. 2, pp. 55–64, 2007. View at: Google Scholar
  35. R. Vitorino, S. De Morais Guedes, R. Ferreira et al., “Two-dimensional electrophoresis study of in vitro pellicle formation and dental caries susceptibility,” European Journal of Oral Sciences, vol. 114, no. 2, pp. 147–153, 2006. View at: Publisher Site | Google Scholar
  36. L. Giusti, C. Baldini, L. Bazzich et al., “Proteome analysis of whole saliva: A new tool for rheumatic diseases - The example of Sjögren's syndrome,” Proteomics, vol. 7, no. 10, pp. 1634–1643, 2007. View at: Publisher Site | Google Scholar
  37. C. Dawes, “Circadian rhythms in human salivary flow rate and composition,” The Journal of Physiology, vol. 220, no. 3, pp. 529–545, 1972. View at: Publisher Site | Google Scholar
  38. M. Castagnola, R. Inzitari, C. Fanali et al., “The Surprising Composition of the Salivary Proteome of Preterm Human Newborn,” Molecular & Cellular Proteomics, vol. 10, no. 1, p. M110.003467, 2010. View at: Publisher Site | Google Scholar
  39. C. Hirtz, F. Chevalier, N. Sommerer et al., “Salivary protein profiling in type 1 diabetes using two-dimensional electrophoresis and mass spectrometry,” Clinical Proteomics, vol. 2, no. 1-2, pp. 117–128, 2006. View at: Publisher Site | Google Scholar
  40. I. Messana, R. Inzitari, C. Fanali, T. Cabras, and M. Castagnola, “Facts and artifacts in proteomics of body fluids. What proteomics of saliva is telling us?” Journal of Separation Science, vol. 31, no. 11, pp. 1948–1963, 2008. View at: Publisher Site | Google Scholar
  41. W. L. Siqueira and C. Dawes, “The salivary proteome: Challenges and perspectives,” Proteomics - Clinical Applications, vol. 5, no. 11-12, pp. 575–579, 2011. View at: Publisher Site | Google Scholar
  42. R. Schipper, A. Loof, J. De Groot, L. Harthoorn, W. Van Heerde, and E. Dransfield, “Salivary protein/peptide profiling with SELDI-TOF-MS,” Annals of the New York Academy of Sciences, vol. 1098, pp. 498–503, 2007. View at: Publisher Site | Google Scholar
  43. B. Ghafouri, C. Tagesson, and M. Lindahl, “Mapping of proteins in human saliva using two-dimensional gel electrophoresis and peptide mass fingerprinting,” Proteomics, vol. 3, no. 6, pp. 1003–1015, 2003. View at: Publisher Site | Google Scholar
  44. M. Hardt, L. R. Thomas, S. E. Dixon et al., “Toward defining the human parotid gland salivary proteome and peptidome: Identification and characterization using 2D SDS-PAGE, ultrafiltration, HPLC, and mass spectrometry,” Biochemistry, vol. 44, no. 8, pp. 2885–2899, 2005. View at: Publisher Site | Google Scholar
  45. S. Hu, P. Denny, P. Denny et al., “Differentially expressed protein markers in human submandibular and sublingual secretions,” International Journal of Oncology, 2004. View at: Publisher Site | Google Scholar
  46. S. Hu, J. Jiang, and D. T. Wong, “Proteomic Analysis of Saliva: 2D Gel Electrophoresis, LC-MS/MS, and Western Blotting,” in Oral Biology, G. J. Seymour, M. P. Cullinan, and N. C. K. Heng, Eds., vol. 666, pp. 31–41, Humana Press, Totowa, NJ, USA, 2010. View at: Publisher Site | Google Scholar
  47. C.-M. Huang, “Comparative proteomic analysis of human whole saliva,” Archives of Oral Biolog, vol. 49, no. 12, pp. 951–962, 2004. View at: Publisher Site | Google Scholar
  48. N. Huq, A. DeAngelis, Z. Rahim, M. U. J. Lucas, K. Cross, and E. Reynolds, “Whole And Parotid Saliva - Protein Profiles As Separated On 5-20% SDS-Polyacrylamide Gradient gel Electrophoresis And Using Maldi-tof Mass Spectrometry,” Annals of Dentistry, vol. 11, no. 1, pp. 24–29, 2004. View at: Publisher Site | Google Scholar
  49. N. L. Huq, K. J. Cross, M. Ung et al., “A review of the salivary proteome and peptidome and saliva-derived peptide therapeutics,” International Journal of Peptide Research and Therapeutics, vol. 13, no. 4, pp. 547–564, 2007. View at: Publisher Site | Google Scholar
  50. I. Messana, T. Cabras, R. Inzitari et al., “Characterization of the human salivary basic proline-rich protein complex by a proteomic approach,” Journal of Proteome Research, vol. 3, no. 4, pp. 792–800, 2004. View at: Publisher Site | Google Scholar
  51. R. Vitorino, M. J. C. Lobo, A. J. Ferrer-Correira et al., “Identification of human whole saliva protein components using proteomics,” Proteomics, vol. 4, no. 4, pp. 1109–1115, 2004. View at: Publisher Site | Google Scholar
  52. Y. Yao, E. A. Berg, C. E. Costello, R. F. Troxler, and F. G. Oppenheim, “Identification of protein components in human acquired enamel pellicle and whole saliva using novel proteomics approaches,” The Journal of Biological Chemistry, vol. 278, no. 7, pp. 5300–5308, 2003. View at: Publisher Site | Google Scholar
  53. M. Castagnola, T. Cabras, F. Iavarone et al., “The human salivary proteome: A critical overview of the results obtained by different proteomic platforms,” Expert Review of Proteomics, vol. 9, no. 1, pp. 33–46, 2012. View at: Publisher Site | Google Scholar
  54. S. K. Al-Tarawneh and S. Bencharit, “Applications of Surface-Enhanced Laser Desorption/Ionization Time-Of- Flight (SELDI-TOF) Mass Spectrometry in Defining Salivary Proteomic Profiles,” The Open Dentistry Journal, vol. 3, no. 1, pp. 74–79, 2009. View at: Publisher Site | Google Scholar
  55. P. Denny and et al., “The Proteomes of Human Parotid and Submandibular/Sublingual Gland Salivas Collected as the Ductal Secretions,” Journal of Proteome Research, vol. 7, no. 5, pp. 1994–2006, 2008. View at: Google Scholar
  56. T. Guo, P. A. Rudnick, W. Wang, C. S. Lee, D. L. Devoe, and B. M. Balgley, “Characterization of the human salivary proteome by capillary isoelectric focusing/nanoreversed-phase liquid chromatography coupled with ESI-tandem MS,” Journal of Proteome Research, vol. 5, no. 6, pp. 1469–1478, 2006. View at: Publisher Site | Google Scholar
  57. H. Xie, N. L. Rhodus, R. J. Griffin, J. V. Carlis, and T. J. Griffin, “A catalogue of human saliva proteins identified by free flow electrophoresis-based peptide separation and tandem mass spectrometry,” Molecular & Cellular Proteomics, vol. 4, no. 11, pp. 1826–1830, 2005. View at: Publisher Site | Google Scholar
  58. R. G. Schipper, E. Silletti, and M. H. Vingerhoeds, “Saliva as research material: Biochemical, physicochemical and practical aspects,” Archives of Oral Biolog, vol. 52, no. 12, pp. 1114–1135, 2007. View at: Publisher Site | Google Scholar
  59. B. Isaksen and M. K. Fagerhol, “Calprotectin inhibits matrix metalloproteinases by sequestration of zinc,” Journal of Clinical Pathology: Molecular Pathology, vol. 54, no. 5, pp. 289–292, 2001. View at: Publisher Site | Google Scholar
  60. C. Kerkhoff, M. Klempt, V. Kaever, and C. Sorg, “The two calcium-binding proteins, S100A8 and S100A9, are involved in the metabolism of arachidonic acid in human neutrophils,” The Journal of Biological Chemistry, vol. 274, no. 46, pp. 32672–32679, 1999. View at: Publisher Site | Google Scholar
  61. L. I. Sakkas, “New developments in the pathogenesis of systemic sclerosis,” Autoimmunity, vol. 38, no. 2, pp. 113–116, 2005. View at: Publisher Site | Google Scholar
  62. P. Madsen, H. H. Rasmussen, H. Leffers et al., “Molecular cloning, occurrence, and expression of a novel partially secreted protein ‘psoriasis’ that is highly up-regulated in psoriatic skin,” Journal of Investigative Dermatology, vol. 97, no. 4, pp. 701–712, 1991. View at: Publisher Site | Google Scholar
  63. O. D. Weiner, G. Servant, M. D. Welch, T. J. Mitchison, J. W. Sedat, and H. R. Bourne, “Spatial control of actin polymerization during neutrophil chemotaxis,” Nature Cell Biology, vol. 1, no. 2, pp. 75–81, 1999. View at: Publisher Site | Google Scholar
  64. H. Watanabe, T. Seino, and Y. Sato, “Antibodies to triosephosphate isomerase in patients with neuropsychiatric lupus,” Biochemical and Biophysical Research Communications, vol. 321, no. 4, pp. 949–953, 2004. View at: Publisher Site | Google Scholar
  65. S.-H. Kim, S. M. Lessner, Y. Sakurai, and Z. S. Galis, “Cyclophilin A as a Novel Biphasic Mediator of Endothelial Activation and Dysfunction,” The American Journal of Pathology, vol. 164, no. 5, pp. 1567–1574, 2004. View at: Publisher Site | Google Scholar
  66. F. U. Hartl and M. Hayer-Hartl, “Molecular chaperones in the cytosol: from nascent chain to folded protein,” Science, vol. 295, no. 5561, pp. 1852–1858, 2002. View at: Publisher Site | Google Scholar
  67. W. L. Siqueira, E. Salih, D. L. Wan, E. J. Helmerhorst, and F. G. Oppenheim, “Proteome of human minor salivary gland secretion,” Journal of Dental Research, vol. 87, no. 5, pp. 445–450, 2008. View at: Publisher Site | Google Scholar

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