형광제자리부합법을 이용한 치은열구세포 내의 치주염 유발 세균의 관찰

Visualization of periodontopathic bacteria within crevicular epithelial cells with fluorescence in situ hybridization

  • 고영경 (가톨릭대학교 의과대학 치과학교실)
  • Ko, Young-Kyung (Department of Dentistry, School of Medicine, The Catholic University)
  • 발행 : 2008.12.31

초록

Purpose: Periodontal pathogens can invade the host tissue. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The objective of this study was to visualize and evaluate presence of Porphyromonas gingivalis and Tannerella forsythia in crevicular epithelial cells of periodontally healthy subjects and chronic periodontitis patients. Materials and Methods: A total of 666 crevicular epithelial cells in the samples obtained from 27 chronic periodontitis patients and 9 healthy volunteers were examined. Specific probes for P. gingivalis and T. forsythia and a universal probe for detection of all eubacteria targeting 168 rRNA for fluorescence in situ hybridization was used in conjunction with confocal laser scanning microscopy. Results: 98.99% of sulcular epithelial cells from healthy volunteers and 84.40% of pocket epithelial cells from periodontitis patients were found to harbor bacteria. P. gingivalis and T. forsythia were discovered more often in crevicular epithelial cells from periodontitis patients. Conclusion: P. gingivalis and T. forsythia can invade crevicular epithelial cells and intracellular bacteria may act as a source of bacteria for persistent infection.

키워드

참고문헌

  1. Noiri Y, Ozaki K, Nakae H, Matsuo T, Ebisu S. An immunohistochemical study on the localization of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus in human periodontal pockets. J Periodontal Res 1997;32:598-607 https://doi.org/10.1111/j.1600-0765.1997.tb00937.x
  2. Rudney JD, Chen R, and Sedgewick GJ. Intracellular Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in buccal epithelial cells collected from human subjects. Infection and Immunity 2001;69:2700-2707 https://doi.org/10.1128/IAI.69.4.2700-2707.2001
  3. Fillery ED, Pekovic DD. Identification of microorganisms in immunopathological mechanism on human gingivitis. J Dent Res 1982;61:25 https://doi.org/10.1177/00220345820610010501
  4. Saglie FR, Marfany A, Carmago P. Intragingival occurrence of Actinobacillus actinomycemtemcomitans and Bacteroides gingivalis in active destructive periodontal lesions. J Periodontol 1988;59:259 https://doi.org/10.1902/jop.1988.59.4.259
  5. Carranza FA, Jr, Saglie R, Newman MG. Scanning and transmission electron microscopic study of tissue-invading microorganisms in localized juvenile periodontitis. J Periodontol 1983;54:598 https://doi.org/10.1902/jop.1983.54.10.598
  6. Sreenivasan PK, Meyer DH, FIves-Tayor PM. Requirements for invasion of epithelial cells by Actinobacillus actimomycetemcomitans. Infect Immun 1993; 61:1239
  7. Han YW, Shi W, Huang GT et al. Interactions between periodontal bacteria and human oral epithelial cells: Fusobacterium nucleatum adheres to and invades epithelial cells. Infect Immun 2000;68:3140 https://doi.org/10.1128/IAI.68.6.3140-3146.2000
  8. Sandros J, Papapanou P, Dahlen G. Porphyromonas gingivalis invades oral epithelial cells in vitro. J Periodontal Res 1993;28:219 https://doi.org/10.1111/j.1600-0765.1993.tb01072.x
  9. Wang B, Holt SC. Interaction of Treponema denticola with HEp-2 cells. J Dent Res 1993;72:324
  10. Madianos PN, Papapanou PN, Nannmark U, Dahlen G, Sandros J. Porphyromonas gingivalis FDC381 multiplies and persistis within the human oral epithelial cells in vitro. Infect Immun 1996;64:660-664
  11. Dale BA. Periodontal epithelium: a newly recognized role in health and disease. Periodontol 2000 2002;30:70-78 https://doi.org/10.1034/j.1600-0757.2002.03007.x
  12. Moter A, Gobel UB. Fluorescence in situ hybridization (FISH) for direct visualization of microorganisms. J Microb Methods 2000;41:85-112 https://doi.org/10.1016/S0167-7012(00)00152-4
  13. Sakamoto M, Umeda M, Benno Y. Molecular analysis of human oral microbiota. J Periodontal Res 2005;40:277-285 https://doi.org/10.1111/j.1600-0765.2005.00793.x
  14. Claridge JE 3rd. Impact of 16S rRNA sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev 2004;17:840-62 https://doi.org/10.1128/CMR.17.4.840-862.2004
  15. Foldes-Papp Z, Demel U, Tilz GP. Laser scanning confocal fluorescence microscopy: an overview. International Immunopharmacology. 2003;3:1715-1729 https://doi.org/10.1016/S1567-5769(03)00140-1
  16. Michalet X, Kapanidis AN, Laurence T, Pinaud F, Doose S, Pflughoefft M, Weiss S. The power and prospects of fluorescence microscopies and spectroscopies. Annu Rev Biophys Biomol Struct 2003;32:161-82 https://doi.org/10.1146/annurev.biophys.32.110601.142525
  17. Dibart S, Skobe Z, Snapp KR et al. Identification of bacterial species on or in crevicular epithelial cells from healthy and periodontally diseased patients using DNA-DNA hybridization. Oral Microbiol Immunol 1998;13:30-35 https://doi.org/10.1111/j.1399-302X.1998.tb00747.x
  18. Colombo AV, Silva CM, Haffajee A, Colombo AP. Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions. J Med Microbiol 2006;55:606-615
  19. Colombo AV, Silva CM, Haffajee A, Colombo APV. Identification of intracellular oral species within human crevicular epithelial cells from subjects with chronic periodontitis by fluorescence in situ hybridization. J Periodontal Res. 2007;42:236-243 https://doi.org/10.1111/j.1600-0765.2006.00938.x
  20. Loy A, Maixner F, Wagner M, Horn M. probeBase - an online resource for rRNA-targeted oligonucleotide probes: new features 2007. Nucleic Acids Res. 2007;35:D800-D804 https://doi.org/10.1093/nar/gkl856
  21. Lamont RJ, Yilmaz o. In or out: the invasiveness of oral bacteria. Periodontol 2000 2002;30:61-69 https://doi.org/10.1034/j.1600-0757.2002.03006.x
  22. Sandros J, Karlsson C, Lappin DF, et al. Cytokine responses of oral epithelial cells to Porphyromonas gingivalis infection. J Dent Res 2000;79:1804-1814
  23. Andrian E, Grenier D, Rouabhia M. Porphrymonas gingivalis-epithelial cell interactions in periodontitis 2006;85: 392-403
  24. Rudney JD, Chen R, Sedgewick GJ. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis are components of a polymicrobial intracellular flora within human buccal cells. J Dent Res 2005;84:59-63 https://doi.org/10.1177/154405910508400110
  25. Johnson JD, Chen R, Lenton PA et al. Persistence of extracrevicular bacterial reservoirs after treatment of aggressive periodontitis. J Periodontol 2008;79:2305-2312 https://doi.org/10.1902/jop.2008.080254