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In Vitro Invasion and Survival of Porphyromonas gingivalis in Gingival Fibroblasts; Role of the Capsule

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Archivum Immunologiae et Therapiae Experimentalis Aims and scope

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.

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Acknowledgments

We thank the “Higher education commission of Pakistan” for supporting this study financially. We thank Dr. Nina Scheres for her valuable technical support and Dr. Jorg Brunner for providing the ΔEpsC isogenic mutant of P. gingivalis W83.

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Authors and Affiliations

  1. 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

    Muhammad Irshad, Wil A. van der Reijden & Wim Crielaard

  2. Section of Periodontology, Department of Conservative and Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Vrije University Amsterdam, Gustav Mahlerlaan 3001, 1081 LA, Amsterdam, The Netherlands

    Marja L. Laine

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  1. Muhammad Irshad
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  2. Wil A. van der Reijden
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Correspondence to Marja L. Laine.

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Irshad, M., van der Reijden, W.A., Crielaard, W. et al. In Vitro Invasion and Survival of Porphyromonas gingivalis in Gingival Fibroblasts; Role of the Capsule. Arch. Immunol. Ther. Exp. 60, 469–476 (2012). https://doi.org/10.1007/s00005-012-0196-8

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  • DOI: https://doi.org/10.1007/s00005-012-0196-8

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