title: In vitro invasion and survival of Porphyromonas gingivalis in gingival fibroblasts; role of the capsule.
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.
authors.affiliation: Section of Preventive Dentistry, Department of Conservative and Preventive Dentistry, Academic Centre for Dentistry Amsterdam, ACTA, University of Amsterdam and Vrije University Amsterdam, Amsterdam, The Netherlands.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Irshad
authors.forename: Wil A
authors.initials: WA
authors.lastname: van der Reijden
authors.forename: Wim
authors.initials: W
authors.lastname: Crielaard
authors.forename: Marja L
authors.initials: ML
authors.lastname: Laine
mesh_terms: D000658:Amoxicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016667:Bacterial Capsules / Q000187:drug effects / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000382:microbiology*; D005881:Gingiva / Q000187:drug effects / Q000382:microbiology*; D006801:Humans; D008795:Metronidazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability; D008875:Middle Aged; D009154:Mutation; D016966:Porphyromonas gingivalis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D008795:Metronidazole; D000658:Amoxicillin