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Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells
Published on: December 17, 2015
Bacterial extracellular vesicles from Porphyromonas gingivalis - proteomic profile and influence on oral fibroblasts
Helene Rygvold Haugsten1, Anne Karin Kristoffersen1, Morten Enersen1
1Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Norway.
Objectives:
In this study, we aimed to explore the protein content of bacterial extracellular vesicles (bEVs) from three strains of Porphyromonas gingivalis, an anaerobic gram-negative bacterium closely associated with periodontitis. Additionally, we aimed to investigate the effects of bEV uptake on human oral fibroblasts, to better understand the role of these vesicles in the pathogenesis of periodontitis.
Design:
Liquid chromatography-mass spectrometry was performed on bEVs from three strains of P. gingivalis with different virulence: ATCC 33277, A7A1-28, and W83. Three key proteins associated with virulence were validated using dot blot. Proliferation and viability of human oral fibroblasts after incubation with bEVs were assessed at different timepoints and different concentrations of bEVs.
Results:
A total of 307 proteins were identified in the bEVs from the three strains of P. gingivalis. Among those were several relevant for virulence, for example gingipains, fimbriae, and peptidyl-arginine deiminase (PPAD). When oral fibroblasts were exposed to bEVs from A7A1-28 or W83, proliferation was significantly affected after 12 and 24 h at one bEV concentration. Furthermore, viability was significantly affected by bEVs from W83 at several timepoints and bEV concentrations. bEVs from ATCC 33277 did not induce any significant changes.
Conclusion:
Several proteins associated with virulence were detected in bEVs from all three strains of P. gingivalis, with varying abundance. Uptake of these bEVs can influence human cells, as exemplified by changes in viability and proliferation.
Insights
Bacterial extracellular vesicles (bEVs) from Porphyromonas gingivalis contain virulence proteins that affect human oral fibroblast proliferation and viability. These findings shed light on the role of bEVs in periodontitis pathogenesis.
Area of Science:
- Microbiology
- Cell Biology
- Periodontal Disease Research
Background:
- Porphyromonas gingivalis is an anaerobic Gram-negative bacterium linked to periodontitis.
- Bacterial extracellular vesicles (bEVs) are implicated in inter-bacterial and host-bacterial communication.
- Understanding bEVs' role in periodontitis pathogenesis is crucial for developing targeted therapies.
Purpose of the Study:
- To characterize the protein cargo of bEVs from three P. gingivalis strains.
- To investigate the impact of P. gingivalis bEVs on human oral fibroblast proliferation and viability.
- To elucidate the contribution of bEVs to periodontitis pathogenesis.
Main Methods:
- Liquid chromatography-mass spectrometry (LC-MS) was used to identify proteins in bEVs from P. gingivalis strains (ATCC 33277, A7A1-28, W83).
- Dot blot assays validated the presence of key virulence proteins.
- Human oral fibroblasts were incubated with bEVs to assess proliferation and viability at various timepoints and concentrations.
Main Results:
- A total of 307 proteins were identified in the bEVs, including virulence factors like gingipains and fimbriae.
- bEVs from A7A1-28 and W83 strains significantly affected fibroblast proliferation and viability.
- bEVs from the ATCC 33277 strain did not induce significant changes in fibroblast behavior.
Conclusions:
- P. gingivalis bEVs harbor diverse virulence-associated proteins.
- Uptake of these bEVs can modulate human oral fibroblast functions, influencing disease progression.
- These findings highlight bEVs as key mediators in the pathogenesis of periodontitis.
