In vitro invasion and survival of Porphyromonas gingivalis in gingival fibroblasts; role of the capsule

Arch Immunol Ther Exp (Warsz). 2012 Dec;60(6):469-76. doi: 10.1007/s00005-012-0196-8. Epub 2012 Sep 5.

Abstract

Porphyromonas gingivalis is a Gram-negative, anaerobic bacterium involved in periodontitis and peri-implantitis that can invade and survive inside host cells in vitro. P. gingivalis can invade human gingival fibroblasts (GF), but no data are available about the role of P. gingivalis' capsule in GF invasion. In the current study, we aimed to determine the ability of three strains of P. gingivalis (encapsulated wild type W83, non-encapsulated HG91 and the non-encapsulated insertional isogenic knockout mutant of W83, ΔEpsC) to invade GF and the ability of internalized P. gingivalis to survive in vitro antibiotic treatment. The ability of P. gingivalis strains to invade GF was tested using an antibiotic protection assay at multiplicity of infection (MOI) 100 and 1000. The survival of internalized P. gingivalis cells was further analyzed by subsequent in vitro treatment with either metronidazole or amoxicillin alone or a combination of metronidazole and amoxicillin and anaerobic culture viability counts. All strains of P. gingivalis used in this study were able to invade GFs. The non-encapsulated mutant of W83 (ΔEpsC mutant) was significantly more invasive than the wild type W83 at MOI 100 (p value 0.025) and MOI 1000 (p value 0.038). Furthermore, internalized P. gingivalis was able to resist in vitro antibiotic treatment. As demonstrated by the differences in invasion efficiencies of P. gingivalis strain W83 and its isogenic mutant ΔEpsC, the capsule of P. gingivalis makes it less efficient in invading gingival fibroblasts. Moreover, internalized P. gingivalis can survive antibiotic treatment in vitro.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amoxicillin / pharmacology
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Capsules / drug effects
  • Bacterial Capsules / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cells, Cultured
  • Female
  • Fibroblasts / drug effects
  • Fibroblasts / microbiology*
  • Gingiva / drug effects
  • Gingiva / microbiology*
  • Humans
  • Metronidazole / pharmacology
  • Microbial Sensitivity Tests
  • Microbial Viability
  • Middle Aged
  • Mutation
  • Porphyromonas gingivalis / drug effects
  • Porphyromonas gingivalis / genetics
  • Porphyromonas gingivalis / metabolism*
  • Porphyromonas gingivalis / pathogenicity

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Metronidazole
  • Amoxicillin