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Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoE Mice.

Chukkapalli SS, Velsko IM, Rivera-Kweh MF, Zheng D, Lucas AR, Kesavalu L - PLoS ONE (2015)

Bottom Line: Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis.Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES.In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, College of Dentistry, University of Florida, Gainesville, Florida, United States of America.

ABSTRACT
Periodontal disease (PD) develops from a synergy of complex subgingival oral microbiome, and is linked to systemic inflammatory atherosclerotic vascular disease (ASVD). To investigate how a polybacterial microbiome infection influences atherosclerotic plaque progression, we infected the oral cavity of ApoE mice with a polybacterial consortium of 4 well-characterized periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, Tannerealla forsythia and Fusobacterium nucleatum, that have been identified in human atherosclerotic plaque by DNA screening. We assessed periodontal disease characteristics, hematogenous dissemination of bacteria, peripheral T cell response, serum inflammatory cytokines, atherosclerosis risk factors, atherosclerotic plaque development, and alteration of aortic gene expression. Polybacterial infections have established gingival colonization in ApoE hyperlipidemic mice and displayed invasive characteristics with hematogenous dissemination into cardiovascular tissues such as the heart and aorta. Polybacterial infection induced significantly higher levels of serum risk factors oxidized LDL (p < 0.05), nitric oxide (p < 0.01), altered lipid profiles (cholesterol, triglycerides, Chylomicrons, VLDL) (p < 0.05) as well as accelerated aortic plaque formation in ApoE mice (p < 0.05). Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis. Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES. In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules. This study demonstrates unique differences in the host immune response to a polybacterial periodontal infection with atherosclerotic lesion progression in a mouse model.

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Viable bacteria were recovered from infected mouse heart and aorta.(A). FISH reveals cluster of coccobacilli morphotype of P. gingivalis in mouse aortic arch at 12 weeks, indicated by white arrow heads. (B). Enlarged inset in D showing clusters of P. gingivalis coccobacilli morphotype indicated by white arrow heads. (C). No bacteria were detected in sham-infected mouse aortic tissues. Scale bar is 10 micrometers.
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pone.0143291.g003: Viable bacteria were recovered from infected mouse heart and aorta.(A). FISH reveals cluster of coccobacilli morphotype of P. gingivalis in mouse aortic arch at 12 weeks, indicated by white arrow heads. (B). Enlarged inset in D showing clusters of P. gingivalis coccobacilli morphotype indicated by white arrow heads. (C). No bacteria were detected in sham-infected mouse aortic tissues. Scale bar is 10 micrometers.

Mentions: Homogenates of heart and aorta were cultured for evidence of viable, invasive bacteria. After 14 days of incubation, colonies were recovered on FAA plates from the aortas (n = 3) and hearts (n = 2) infected mice sacrificed at 24 weeks. Colonies were brown indicating heme-accumulation, with a shiny, mucoid appearance. Colonies were Gram-stained, and revealed Gram-negative species of varying morphotypes (coccobacilli, short bacilli, long slender bacilli). Colonies were tested for the presence of P. gingivalis, T. denticola, T. forsythia, and F. nucleatum by PCR. Colonies recovered from the aorta of one mouse were positive for both T. denticola and F. nucleatum (S1 Table), while a different colony from the aorta and one colony from the heart of the same mouse tested positive for T. denticola (S1 Table). Colonies recovered from the aorta of a second mouse and the heart of a third mouse were found positive for F. nucleatum by 16S rRNA species specific PCR. Recovering viable oral bacteria from the heart and aorta of mice at sacrifice 14 days after the last oral inoculation demonstrates chronic exposure to and colonization of the bacteria, and their potential to actively invade and persist in vascular tissues. Additionally, FISH was performed on aorta sections of polybacterial-infected and sham-infected mice to detect viable invasive bacteria in situ [21]. P. gingivalis were detected in the aortic wall of one 12 week-infected mouse (Fig 3A and 3B), demonstrating the potential to actively invade and colonize the vasculature. No bacteria were detected in sham-infected mouse aortic tissues (Fig 3C).


Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoE Mice.

Chukkapalli SS, Velsko IM, Rivera-Kweh MF, Zheng D, Lucas AR, Kesavalu L - PLoS ONE (2015)

Viable bacteria were recovered from infected mouse heart and aorta.(A). FISH reveals cluster of coccobacilli morphotype of P. gingivalis in mouse aortic arch at 12 weeks, indicated by white arrow heads. (B). Enlarged inset in D showing clusters of P. gingivalis coccobacilli morphotype indicated by white arrow heads. (C). No bacteria were detected in sham-infected mouse aortic tissues. Scale bar is 10 micrometers.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664240&req=5

pone.0143291.g003: Viable bacteria were recovered from infected mouse heart and aorta.(A). FISH reveals cluster of coccobacilli morphotype of P. gingivalis in mouse aortic arch at 12 weeks, indicated by white arrow heads. (B). Enlarged inset in D showing clusters of P. gingivalis coccobacilli morphotype indicated by white arrow heads. (C). No bacteria were detected in sham-infected mouse aortic tissues. Scale bar is 10 micrometers.
Mentions: Homogenates of heart and aorta were cultured for evidence of viable, invasive bacteria. After 14 days of incubation, colonies were recovered on FAA plates from the aortas (n = 3) and hearts (n = 2) infected mice sacrificed at 24 weeks. Colonies were brown indicating heme-accumulation, with a shiny, mucoid appearance. Colonies were Gram-stained, and revealed Gram-negative species of varying morphotypes (coccobacilli, short bacilli, long slender bacilli). Colonies were tested for the presence of P. gingivalis, T. denticola, T. forsythia, and F. nucleatum by PCR. Colonies recovered from the aorta of one mouse were positive for both T. denticola and F. nucleatum (S1 Table), while a different colony from the aorta and one colony from the heart of the same mouse tested positive for T. denticola (S1 Table). Colonies recovered from the aorta of a second mouse and the heart of a third mouse were found positive for F. nucleatum by 16S rRNA species specific PCR. Recovering viable oral bacteria from the heart and aorta of mice at sacrifice 14 days after the last oral inoculation demonstrates chronic exposure to and colonization of the bacteria, and their potential to actively invade and persist in vascular tissues. Additionally, FISH was performed on aorta sections of polybacterial-infected and sham-infected mice to detect viable invasive bacteria in situ [21]. P. gingivalis were detected in the aortic wall of one 12 week-infected mouse (Fig 3A and 3B), demonstrating the potential to actively invade and colonize the vasculature. No bacteria were detected in sham-infected mouse aortic tissues (Fig 3C).

Bottom Line: Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis.Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES.In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, College of Dentistry, University of Florida, Gainesville, Florida, United States of America.

ABSTRACT
Periodontal disease (PD) develops from a synergy of complex subgingival oral microbiome, and is linked to systemic inflammatory atherosclerotic vascular disease (ASVD). To investigate how a polybacterial microbiome infection influences atherosclerotic plaque progression, we infected the oral cavity of ApoE mice with a polybacterial consortium of 4 well-characterized periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, Tannerealla forsythia and Fusobacterium nucleatum, that have been identified in human atherosclerotic plaque by DNA screening. We assessed periodontal disease characteristics, hematogenous dissemination of bacteria, peripheral T cell response, serum inflammatory cytokines, atherosclerosis risk factors, atherosclerotic plaque development, and alteration of aortic gene expression. Polybacterial infections have established gingival colonization in ApoE hyperlipidemic mice and displayed invasive characteristics with hematogenous dissemination into cardiovascular tissues such as the heart and aorta. Polybacterial infection induced significantly higher levels of serum risk factors oxidized LDL (p < 0.05), nitric oxide (p < 0.01), altered lipid profiles (cholesterol, triglycerides, Chylomicrons, VLDL) (p < 0.05) as well as accelerated aortic plaque formation in ApoE mice (p < 0.05). Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis. Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES. In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules. This study demonstrates unique differences in the host immune response to a polybacterial periodontal infection with atherosclerotic lesion progression in a mouse model.

Show MeSH
Related in: MedlinePlus