Advertisement

The Journal of Microbiology

, Volume 50, Issue 1, pp 143–148 | Cite as

Effects of nicotine on the growth and protein expression of Porphyromonas gingivalis

  • Orson Baek
  • Weidong Zhu
  • Hyeong C. Kim
  • Seok-Woo LeeEmail author
Articles

Abstract

Tobacco smoking is considered one of the most significant environmental risk factors for destructive periodontal disease. The effect of smoking on periodontopathic microbiota has not yet been elucidated, as previous studies failed to identify a concrete relationship between periodontopathic microorganisms and smoking. However, it is likely that smoking, as an environmental stress factor, may affect the behavior of dental plaque microorganisms, ultimately leading to alteration of the host-parasite interaction. The goal of this study was to examine the effect of nicotine, a major component of tobacco, on the growth and protein expression of the crucial periodontal pathogen Porphyromonas gingivalis. The growth of P. gingivalis 381 was measured after bacterial cells were cultivated in liquid broth containing various nicotine concentrations. First, P. gingivalis cells were allowed to grow in the presence of a single dose of nicotine (the single exposure protocol) at 0, 1, 2, 4, and 8 mg/L, respectively. Second, P. gingivalis cells were exposed to five consecutive doses of nicotine (the multiple exposure protocol) at 0, 1, 2, and 4 mg/L, respectively. Bacterial growth was measured by optical density and protein expression was analyzed by SDS-PAGE and 2-D gel electrophoresis. In the single nicotine exposure protocol, it was observed that the growth of P. gingivalis 381 was inhibited by nicotine in a dose-dependent manner. In the multiple nicotine exposure protocol, the growth rate of P. gingivalis increased with each subsequent nicotine exposure, even though bacterial growth was also inhibited in a dose dependent fashion. SDS-PAGE and 2-D gel electrophoresis analyses revealed a minor change in the pattern of protein expression, showing differences in proteins with low molecular weights (around 20 kDa) on exposure to nicotine. The results of this study suggest that nicotine exerts an inhibitory effect on the growth of P. gingivalis, and has a potential to modulate protein expression in P. gingivalis.

Keywords

smoking nicotine protein expression virulence periodontal disease oral anaerobic bacterium 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Albandar, J.M. 2002. Global risk factors and risk indicators for periodontal diseases. Periodontol. 2000 29, 177–206.PubMedCrossRefGoogle Scholar
  2. Amano, A., Sharma, A., Sojar, H.T., Kuramitsu, H.K., and Genco, R.J. 1994. Effects of temperature stress on expression of fimbriae and superoxide dismutase by Porphyromonas gingivalis. Infect. Immun. 62, 4682–4685.PubMedGoogle Scholar
  3. Bagaitkar, J., Demuth, D.R., Daep, C.A., Renaud, D.E., Pierce, D.L., and Scott, D.A. 2010. Tobacco upregulates P. gingivalis fimbrial proteins which induce TLR2 hyposensitivity. PLoS One 5, e9323.PubMedCrossRefGoogle Scholar
  4. Bagaitkar, J., Williams, L.R., Renaud, D.E., Bemakanakere, M.R., Martin, M., Scott, D.A., and Demuth, D.R. 2009. Tobacco-induced alterations to Porphyromonas gingivalis-host interactions. Environ. Microbiol. 11, 1242–1253.PubMedCrossRefGoogle Scholar
  5. Benowitz, N.L. and Jacob, P.3rd. 1984. Daily intake of nicotine during cigarette smoking. Clin. Pharmacol. Ther. 35, 499–504.PubMedCrossRefGoogle Scholar
  6. Bergstrom, J. and Preber, H. 1986. The influence of cigarette smoking on the development of experimental gingivitis. J. Periodontal. Res. 21, 668–676.PubMedCrossRefGoogle Scholar
  7. Bostrom, L., Bergstrom, J., Dahlen, G., and Linder, L.E. 2001. Smoking and subgingival microflora in periodontal disease. J. Clin. Periodontol. 28, 212–219.PubMedCrossRefGoogle Scholar
  8. Cogo, K., Calvi, B.M., Mariano, F.S., Franco, G.C., Goncalves, R.B., and Groppo, F.C. 2009. The effects of nicotine and cotinine on Porphyromonas gingivalis colonisation of epithelial cells. Arch. Oral Biol. 54, 1061–1067.PubMedCrossRefGoogle Scholar
  9. Darby, I.B., Hodge, P.J., Riggio, M.P., and Kinane, D.F. 2000. Microbial comparison of smoker and non-smoker adult and early-onset periodontitis patients by polymerase chain reaction. J. Clin. Periodontol. 27, 417–424.PubMedCrossRefGoogle Scholar
  10. Finlay, B.B. and Falkow, S. 1989. Common themes in microbial pathogenicity. Microbiol. Rev. 53, 210–230.PubMedGoogle Scholar
  11. Finlay, B.B. and Falkow, S. 1997. Common themes in microbial pathogenicity revisited. Microbiol. Mol. Biol. Rev. 61, 136–169.PubMedGoogle Scholar
  12. Forng, R.Y., Champagne, C., Simpson, W., and Genco, C.A. 2000. Environmental cues and gene expression in Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. Oral Dis. 6, 351–365.PubMedCrossRefGoogle Scholar
  13. Holt, S.C. and Ebersole, J.L. 2005. Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia: the “red complex”, a prototype polybacterial pathogenic consortium in periodontitis. Periodontol. 2000 38, 72–122.PubMedCrossRefGoogle Scholar
  14. Kamma, J.J., Nakou, M., and Baehni, P.C. 1999. Clinical and microbiological characteristics of smokers with early onset periodontitis. J. Periodontal Res. 34, 25–33.PubMedCrossRefGoogle Scholar
  15. Kinane, D.F. and Hart, T.C. 2003. Genes and gene polymorphisms associated with periodontal disease. Crit. Rev. Oral Biol. Med. 14, 430–449.PubMedCrossRefGoogle Scholar
  16. Kroes, I., Lepp, P.W., and Relman, D.A. 1999. Bacterial diversity within the human subgingival crevice. Proc. Natl. Acad. Sci. USA 96, 14547–14552.PubMedCrossRefGoogle Scholar
  17. Loos, B.G., John, R.P., and Laine, M.L. 2005. Identification of genetic risk factors for periodontitis and possible mechanisms of action. J. Clin. Periodontol. 32Suppl 6, 159–179.PubMedCrossRefGoogle Scholar
  18. McGuire, J.R., McQuade, M.J., Rossmann, J.A., Garnick, J.J., Sutherland, D.E., Scheidt, M.J., and Van Dyke, T.E. 1989. Cotinine in saliva and gingival crevicular fluid of smokers with periodontal disease. J. Periodontol. 60, 176–181.PubMedGoogle Scholar
  19. McKee, A.S., McDermid, A.S., Baskerville, A., Dowsett, A.B., Ellwood, D.C., and Marsh, P.D. 1986. Effect of hemin on the physiology and virulence of Bacteroides gingivalis W50. Infect. Immun. 52, 349–355.PubMedGoogle Scholar
  20. Mekalanos, J.J. 1992. Environmental signals controlling expression of virulence determinants in bacteria. J. Bacteriol. 174, 1–7.PubMedGoogle Scholar
  21. O’Brien-Simpson, N.M., Veith, P.D., Dashper, S.G., and Reynolds, E.C. 2004. Antigens of bacteria associated with periodontitis. Periodontol. 2000 35, 101–134.PubMedCrossRefGoogle Scholar
  22. O’Farrell, P.H. 1975. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021.PubMedGoogle Scholar
  23. Obeid, P. and Bercy, P. 2000. Effects of smoking on periodontal health: a review. Adv. Ther. 17, 230–237.PubMedCrossRefGoogle Scholar
  24. Olczak, T., Simpson, W., Liu, X., and Genco, C.A. 2005. Iron and heme utilization in Porphyromonas gingivalis. FEMS Microbiol. Rev. 29, 119–144.PubMedCrossRefGoogle Scholar
  25. Page, R.C., Offenbacher, S., Schroeder, H.E., Seymour, G.J., and Kornman, K.S. 1997. Advances in the pathogenesis of periodontitis: summary of developments, clinical implications and future directions. Periodontol. 2000 14, 216–248.PubMedCrossRefGoogle Scholar
  26. Palmer, R.M., Wilson, R.F., Hasan, A.S., and Scott, D.A. 2005. Mechanisms of action of environmental factors-tobacco smoking 1. J. Clin. Periodontol. 32Suppl 6, 180–195.PubMedCrossRefGoogle Scholar
  27. Paster, B.J., Boches, S.K., Galvin, J.L., Ericson, R.E., Lau, C.N., Levanos, V.A., Sahasrabudhe, A., and Dewhirst, F.E. 2001. Bacterial diversity in human subgingival plaque. J. Bacteriol. 183, 3770–3783.PubMedCrossRefGoogle Scholar
  28. Percival, R.S., Marsh, P.D., Devine, D.A., Rangarajan, M., duse-Opoku, J., Shepherd, P., and Curtis, M.A. 1999. Effect of temperature on growth, hemagglutination, and protease activity of Porphyromonas gingivalis. Infect. Immun. 67, 1917–1921.PubMedGoogle Scholar
  29. Pridmore, A.M., Devine, D.A., Bonass, W.A., and Silley, P. 1999. Influence of sample preparation technique on two-dimensional gel electrophoresis of proteins from Porphyromonas gingivalis. Lett. Appl. Microbiol. 28, 245–249.PubMedCrossRefGoogle Scholar
  30. Rivera-Hidalgo, F. 2003. Smoking and periodontal disease. Periodontol. 2000. 32, 50–58.PubMedCrossRefGoogle Scholar
  31. Tonetti, M.S. 1998. Cigarette smoking and periodontal diseases: etiology and management of disease. Ann. Periodontol. 3, 88–101.PubMedCrossRefGoogle Scholar
  32. Umeda, M., Chen, C., Bakker, I., Contreras, A., Morrison, J.L., and Slots, J. 1998. Risk indicators for harboring periodontal pathogens. J. Periodontol. 69, 1111–1118.PubMedGoogle Scholar
  33. Van der Velden, U., Varoufaki, A., Hutter, J.W., Xu, L., Timmerman, M.F., Van Winkelhoff, A.J., and Loos, B.G. 2003. Effect of smoking and periodontal treatment on the subgingival microflora. J. Clin. Periodontol. 30, 603–610.PubMedCrossRefGoogle Scholar
  34. Wang, H., Kachman, M.T., Schwartz, D.R., Cho, K.R., and Lubman, D.M. 2002. A protein molecular weight map of ES2 clear cell ovarian carcinoma cells using a two-dimensional liquid separations/mass mapping technique. Electrophoresis 23, 3168–3181.PubMedCrossRefGoogle Scholar
  35. Williams, R.C. 1990. Periodontal disease. N. Engl. J. Med. 322, 373–382.PubMedCrossRefGoogle Scholar
  36. Xie, H., Cai, S., and Lamont, R.J. 1997. Environmental regulation of fimbrial gene expression in Porphyromonas gingivalis. Infect. Immun. 65, 2265–2271.PubMedGoogle Scholar
  37. Zambon, J.J., Grossi, S.G., Machtei, E.E., Ho, A.W., Dunford, R., and Genco, R.J. 1996. Cigarette smoking increases the risk for subgingival infection with periodontal pathogens. J. Periodontol. 67, 1050–1054.PubMedGoogle Scholar

Copyright information

© The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Orson Baek
    • 1
  • Weidong Zhu
    • 2
  • Hyeong C. Kim
    • 1
  • Seok-Woo Lee
    • 3
    • 4
    Email author
  1. 1.Division of Periodontics, College of Dental MedicineColumbia UniversityNew YorkUSA
  2. 2.Department of Medicine, School of MedicineUniversity of California, Los AngelesLos AngelesUSA
  3. 3.Division of Periodontics, School of DentistryMarquette UniversityMilwaukeeUSA
  4. 4.Departments of Periodontics and Dental Education, Dental Science Research Institute, School of DentistryChonnam National UniversityGwangjuRepublic of Korea

Personalised recommendations