[go: up one dir, main page]
More Web Proxy on the site http://driver.im/ Skip to main content

Advertisement

Springer Nature Link
Log in

Sterile-filtered saliva is a strong inducer of IL-6 and IL-8 in oral fibroblasts

  • Original Article
  • Published:
Clinical Oral Investigations Aims and scope Submit manuscript

Abstract

Objectives

Saliva has been implicated to support oral wound healing, a process that requires a transient inflammatory reaction. However, definitive proof that saliva can provoke an inflammatory response remained elusive.

Materials and methods

We investigated the ability of freshly harvested and sterile-filtered saliva to cause an inflammatory response of oral fibroblasts and epithelial cells. The expression of cytokines and chemokines was assessed by microarray, RT-PCR, immunoassays, and Luminex technology. The involvement of signaling pathways was determined by Western blot analysis and pharmacologic inhibitors.

Results

We report that sterile-filtered whole saliva was a potent inducer of IL-6 and IL-8 in fibroblasts from the gingiva, the palate, and the periodontal ligament, but not of oral epithelial cells. This strong inflammatory response requires nuclear factor-kappa B and mitogen-activated protein kinase signaling. The pro-inflammatory capacity is heat stable and has a molecular weight of <40 kDa. Genome-wide microarrays and Luminex technology further revealed that saliva substantially increased expression of other inflammatory genes and various chemokines. To preclude that the observed pro-inflammatory activity is the result of oral bacteria, sterile-filtered parotid saliva, collected under almost aseptic conditions, was used and also increased IL-6 and IL-8 expression in gingiva fibroblasts. The inflammatory response was, furthermore, independent of MYD88, an adapter protein of the Toll-like receptor signaling pathway.

Conclusions

We conclude that saliva can provoke a robust inflammatory response in oral fibroblasts involving the classical nuclear factor-kappa B and mitogen-activated protein kinase signaling pathway.

Clinical relevance

Since fibroblasts but not epithelial cells show a strong inflammatory response, saliva may support the innate immunity of defect sites exposing the oral connective tissue.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
£29.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (United Kingdom)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. du Toit DF, Nortje C (2004) Salivary glands: applied anatomy and clinical correlates. SADJ 59:65–66, 69-71, 73-4

    PubMed  Google Scholar 

  2. Cheaib Z, Lussi A (2011) Impact of acquired enamel pellicle modification on initial dental erosion. Caries Res 45:107–112. doi:10.1159/000324803

    Article  PubMed  Google Scholar 

  3. Jakubovics NS, Kolenbrander PE (2010) The road to ruin: the formation of disease-associated oral biofilms. Oral Dis 16:729–739. doi:10.1111/j.1601-0825.2010.01701.x

    Article  PubMed  Google Scholar 

  4. Yao Y, Berg EA, Costello CE, Troxler RF, Oppenheim FG (2003) Identification of protein components in human acquired enamel pellicle and whole saliva using novel proteomics approaches. J Biol Chem 278:5300–5308. doi:10.1074/jbc.M206333200

    Article  PubMed  Google Scholar 

  5. Nobbs AH, Jenkinson HF, Jakubovics NS (2011) Stick to your gums: mechanisms of oral microbial adherence. J Dent Res 90:1271–1278. doi:10.1177/0022034511399096

    Article  PubMed Central  PubMed  Google Scholar 

  6. Bodner L, Knyszynski A, Adler-Kunin S, Danon D (1991) The effect of selective desalivation on wound healing in mice. Exp Gerontol 26:357–363

    Article  PubMed  Google Scholar 

  7. Bodner L, Dayan D, Rothchild D, Hammel I (1991) Extraction wound healing in desalivated rats. J Oral Pathol Med 20:176–178

    Article  PubMed  Google Scholar 

  8. Grossman N, Binyamin LA, Bodner L (2004) Effect of rat salivary glands extracts on the proliferation of cultured skin cells—a wound healing model. Cell Tissue Bank 5:205–212. doi:10.1007/s10561-004-4367-7

    PubMed  Google Scholar 

  9. Proksch S, Steinberg T, Keller C, Wolkewitz M, Wiedmann-Al-Ahmad M, Finkenzeller G, Hannig C, Hellwig E, Al-Ahmad A (2012) Human saliva exposure modulates bone cell performance in vitro. Clin Oral Investig 16:69–77. doi:10.1007/s00784-010-0506-7

    Article  PubMed  Google Scholar 

  10. Komine K, Kuroishi T, Ozawa A, Komine Y, Minami T, Shimauchi H, Sugawara S (2007) Cleaved inflammatory lactoferrin peptides in parotid saliva of periodontitis patients. Mol Immunol 44:1498–1508. doi:10.1016/j.molimm.2006.09.003

    Article  PubMed  Google Scholar 

  11. Ohshima M, Sato M, Ishikawa M, Maeno M, Otsuka K (2002) Physiologic levels of epidermal growth factor in saliva stimulate cell migration of an oral epithelial cell line, HO-1-N-1. Eur J Oral Sci 110:130–136

    Article  PubMed  Google Scholar 

  12. Humphreys-Beher MG, Macauley SP, Chegini N, van Setten G, Purushotham K, Stewart C, Wheeler TT, Schultz GS (1994) Characterization of the synthesis and secretion of transforming growth factor-alpha from salivary glands and saliva. Endocrinology 134:963–970

    PubMed  Google Scholar 

  13. Oudhoff MJ, Bolscher JG, Nazmi K, Kalay H, van ’t Hof W, Amerongen AV, Veerman EC (2008) Histatins are the major wound-closure stimulating factors in human saliva as identified in a cell culture assay. FASEB J 22:3805–3812. doi:10.1096/fj.08-112003

    Article  PubMed  Google Scholar 

  14. Kato T, Imatani T, Minaguchi K, Saitoh E, Okuda K (2002) Salivary cystatins induce interleukin-6 expression via cell surface molecules in human gingival fibroblasts. Mol Immunol 39:423–430

    Article  PubMed  Google Scholar 

  15. Pradhan L, Cai X, Wu S, Andersen ND, Martin M, Malek J, Guthrie P, Veves A, Logerfo FW (2011) Gene expression of pro-inflammatory cytokines and neuropeptides in diabetic wound healing. J Surg Res 167:336–342. doi:10.1016/j.jss.2009.09.012

    Article  PubMed  Google Scholar 

  16. Papanicolaou DA, Wilder RL, Manolagas SC, Chrousos GP (1998) The pathophysiologic roles of interleukin-6 in human disease. Ann Intern Med 128:127–137

    Article  PubMed  Google Scholar 

  17. Baggiolini M, Walz A, Kunkel SL (1989) Neutrophil-activating peptide-1/interleukin 8, a novel cytokine that activates neutrophils. J Clin Invest 84:1045–1049. doi:10.1172/JCI114265

    Article  PubMed Central  PubMed  Google Scholar 

  18. Scheres N, Laine ML, de Vries TJ, Everts V, van Winkelhoff AJ (2010) Gingival and periodontal ligament fibroblasts differ in their inflammatory response to viable Porphyromonas gingivalis. J Periodontal Res 45:262–270. doi:10.1111/j.1600-0765.2009.01229.x

    Article  PubMed  Google Scholar 

  19. Uehara A, Takada H (2007) Functional TLRs and NODs in human gingival fibroblasts. J Dent Res 86:249–254

    Article  PubMed  Google Scholar 

  20. Gornowicz A, Bielawska A, Bielawski K, Grabowska SZ, Wojcicka A, Zalewska M, Maciorkowska E (2012) Pro-inflammatory cytokines in saliva of adolescents with dental caries disease. Ann Agric Environ Med AAEM 19:711–716

    Google Scholar 

  21. Ertugrul AS, Sahin H, Dikilitas A, Alpaslan N, Bozoglan A (2013) Comparison of CCL28, interleukin-8, interleukin-1beta and tumor necrosis factor-alpha in subjects with gingivitis, chronic periodontitis and generalized aggressive periodontitis. J Periodontal Res 48:44–51. doi:10.1111/j.1600-0765.2012.01500.x

    Article  PubMed  Google Scholar 

  22. Scott AE, Milward M, Linden GJ, Matthews JB, Carlile MJ, Lundy FT, Naeeni MA, Lorraine Martin S, Walker B, Kinane D, Brock GR, Chapple IL (2012) Mapping biological to clinical phenotypes during the development (21 days) and resolution (21 days) of experimental gingivitis. J Clin Periodontol 39:123–131. doi:10.1111/j.1600-051X.2011.01825.x

    Article  PubMed  Google Scholar 

  23. Teles RP, Likhari V, Socransky SS, Haffajee AD (2009) Salivary cytokine levels in subjects with chronic periodontitis and in periodontally healthy individuals: a cross-sectional study. J Periodontal Res 44:411–417. doi:10.1111/j.1600-0765.2008.01119.x

    Article  PubMed Central  PubMed  Google Scholar 

  24. Severino VO, Napimoga MH, de Lima Pereira SA (2011) Expression of IL-6, IL-10, IL-17 and IL-8 in the peri-implant crevicular fluid of patients with peri-implantitis. Arch Oral Biol 56:823–828. doi:10.1016/j.archoralbio.2011.01.006

    Article  PubMed  Google Scholar 

  25. Groschl M (2009) The physiological role of hormones in saliva. Bioessays 31:843–852. doi:10.1002/bies.200900013

    Article  PubMed  Google Scholar 

  26. Services USDoHaH (2004) Guidance for industry, sterile drug products produced by aseptic processing—current good manufacturing practice. FDA, Rockville

    Google Scholar 

  27. Jin J, Sundararaj KP, Samuvel DJ, Zhang X, Li Y, Lu Z, Lopes-Virella MF, Huang Y (2012) Different signaling mechanisms regulating IL-6 expression by LPS between gingival fibroblasts and mononuclear cells: seeking the common target. Clin Immunol 143:188–199. doi:10.1016/j.clim.2012.01.019

    Article  PubMed Central  PubMed  Google Scholar 

  28. Hwang YS, Lee SK, Park KK, Chung WY (2012) Secretion of IL-6 and IL-8 from lysophosphatidic acid-stimulated oral squamous cell carcinoma promotes osteoclastogenesis and bone resorption. Oral Oncol 48:40–48. doi:10.1016/j.oraloncology.2011.08.022

    Article  PubMed  Google Scholar 

  29. Tang CH, Hsu CJ, Yang WH, Fong YC (2010) Lipoteichoic acid enhances IL-6 production in human synovial fibroblasts via TLR2 receptor, PKCdelta and c-Src dependent pathways. Biochem Pharmacol 79:1648–1657. doi:10.1016/j.bcp.2010.01.025

    Article  PubMed  Google Scholar 

  30. Thomadaki K, Helmerhorst EJ, Tian N, Sun X, Siqueira WL, Walt DR, Oppenheim FG (2011) Whole-saliva proteolysis and its impact on salivary diagnostics. J Dent Res 90:1325–1330. doi:10.1177/0022034511420721

    Article  PubMed Central  PubMed  Google Scholar 

  31. Miyazono K, Hellman U, Wernstedt C, Heldin CH (1988) Latent high molecular weight complex of transforming growth factor beta 1. Purification from human platelets and structural characterization. J Biol Chem 263:6407–6415

    PubMed  Google Scholar 

  32. Takanashi N, Tomosada Y, Villena J, Murata K, Takahashi T, Chiba E, Tohno M, Shimazu T, Aso H, Suda Y, Ikegami S, Itoh H, Kawai Y, Saito T, Alvarez S, Kitazawa H (2013) Advanced application of bovine intestinal epithelial cell line for evaluating regulatory effect of lactobacilli against heat-killed enterotoxigenic Escherichia coli-mediated inflammation. BMC Microbiol 13:54. doi:10.1186/1471-2180-13-54

    Article  PubMed Central  PubMed  Google Scholar 

  33. Yan W, Apweiler R, Balgley BM, Boontheung P, Bundy JL, Cargile BJ, Cole S, Fang X, Gonzalez-Begne M, Griffin TJ, Hagen F, Hu S, Wolinsky LE, Lee CS, Malamud D, Melvin JE, Menon R, Mueller M, Qiao R, Rhodus NL, Sevinsky JR, States D, Stephenson JL, Than S, Yates JR, Yu W, Xie H, Xie Y, Omenn GS, Loo JA, Wong DT (2009) Systematic comparison of the human saliva and plasma proteomes. Proteomics Clin Appl 3:116–134. doi:10.1002/prca.200800140

    Article  PubMed Central  PubMed  Google Scholar 

  34. Loo JA, Yan W, Ramachandran P, Wong DT (2010) Comparative human salivary and plasma proteomes. J Dent Res 89:1016–1023. doi:10.1177/0022034510380414

    Article  PubMed Central  PubMed  Google Scholar 

  35. Odqvist L, Sanchez-Beato M, Montes-Moreno S, Martin-Sanchez E, Pajares R, Sanchez-Verde L, Ortiz-Romero PL, Rodriguez J, Rodriguez-Pinilla SM, Iniesta-Martinez F, Solera-Arroyo JC, Ramos-Asensio R, Flores T, Palanca JM, Bragado FG, Franjo PD, Piris MA (2013) NIK controls classical and alternative NF-kappaB activation and is necessary for the survival of human T-cell lymphoma cells. Clin Cancer Res Off J Am Assoc Cancer Res 19:2319–2330. doi:10.1158/1078-0432.CCR-12-3151

    Article  Google Scholar 

  36. Abend JR, Uldrick T, Ziegelbauer JM (2010) Regulation of tumor necrosis factor-like weak inducer of apoptosis receptor protein (TWEAKR) expression by Kaposi’s sarcoma-associated herpesvirus microRNA prevents TWEAK-induced apoptosis and inflammatory cytokine expression. J Virol 84:12139–12151. doi:10.1128/JVI.00884-10

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Catherine Solioz for skillful technique assistance and Fumie Saji for sharing her research experience with us. We also thank Dr. Jordi Caballé Serrano and Dr. Jianbo Peng for their essential help with the fluorescence microscopy and the Western blot analysis.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Preventive, Restorative and Pediatric Dentistry, School of Dental Medicine, University of Bern, Bern, Switzerland

    Barbara Cvikl, Adrian Lussi & Reinhard Gruber

  2. Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland

    Barbara Cvikl & Reinhard Gruber

  3. Department of Conservative Dentistry and Periodontology, Medical University of Vienna, Vienna, Austria

    Barbara Cvikl & Andreas Moritz

  4. Department of Periodontology, School of Dental Medicine, University of Bern, Bern, Switzerland

    Anton Sculean & Reinhard Gruber

Authors
  1. Barbara Cvikl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Adrian Lussi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andreas Moritz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anton Sculean
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Reinhard Gruber
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Reinhard Gruber.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cvikl, B., Lussi, A., Moritz, A. et al. Sterile-filtered saliva is a strong inducer of IL-6 and IL-8 in oral fibroblasts. Clin Oral Invest 19, 385–399 (2015). https://doi.org/10.1007/s00784-014-1232-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-014-1232-3

Keywords

Search

Navigation