Bioclinical and periodontal characteristics of the population studied.

<p>The Wilcoxon rank sum test was used to compare medians between groups, and the Fisher exact test to compare proportions. *p<0.05. ^†p<0.01. PI: Plaque Index, GI: Gingival Index, PPD: Pocket Probing Depth, CAL: Clinical Attachment Loss, CRP: C-Reactive Protein.</p>(1)<p>Smoking status: never versus former and current.</p

Adjusted logistic regression models for the association of severity of periodontitis and concentrations of inflammatory biomarkers.

<p>The Model A is adjusted for age, gender and smoking (never versus former and current) and the Model B is adjusted for age, gender, smoking and diabetes (n = 32).</p>*<p>p<0.05, OR: Odds Ratio, CI: Confidence Interval.</p

<i>P. gingivalis</i> (<i>Pg</i>) infection promoted neutrophil recruitment, NET formation and inhibited healing.

<p>(A) Hematoxylin/Eosin staining showing the presence of a thrombus and neutrophil accumulation at its luminal pole (right, inset) in <i>Pg</i>-infected rats. The presence of mesenchymatous cells is observed in saline-injected rats (left, inset). (B) Masson's trichrome staining. Fibrosis associated with healing is observed in green in saline-injected rats whereas red staining highlights the presence of a thrombus in <i>Pg</i>-infected rats. (C) Immunostaining for histone H1 (red), nuclei appear in blue (DAPI). Merged images show the presence of extracellular H1 associated with disorganized DNA (inset), but also intact neutrophils characterized by their multilobed nuclei (bottom, right).</p

Increased cell-free DNA (cf-DNA) in the conditioned medium and in plasma of human AAA.

<p>(A) The concentration of cf-DNA was determined in the conditioned medium obtained from the arterial wall of control and aneurysmal aortas, from AAA thrombus and (B) in plasma of healthy subjects and patients with a small (<5 cm) or a large (>5 cm) AAA. Results are presented as box plots in which the median is shown. **p<0.01; ***p<0.0001 (Mann-Whitney analysis). The correlation (n = 32) between AAA diameter, thrombus volume and cf-DNA concentration were obtained by the Least Squares method.</p

Increased MMP9 activity and MPO released by AAA samples of <i>P. gingivalis</i> (<i>Pg</i>) infected rats.

<p>(A) gelatin zymography analysis of saline- and <i>Pg</i>-injected rats (respectively rats R1,2,3 and R4,5,6). MW: molecular weight, Ref: reference containing pro- and active MMP-9. Graphs represent spatial density quantification of pro- and active MMP9 lysis areas (Image J software). (B) MPO concentration was determined by ELISA in conditioned medium and in plasma. *p<0.05, **p<0.01 (Mann-Whitney Analysis).</p

Detection of bacteria in human AAA samples.

<p>(A) Endotoxin levels from gram-negative bacteria were quantified in the conditioned medium of the Intra-luminal thrombus (ILT) and the arterial wall of AAA (n = 16) and control aortas (n = 10) using the Limulus Amebocyte Lysate chromogenic assay kit. *p<0.05; **p<0.01 (Mann-Whitney Analysis). DNA was extracted from the ILT and associated arterial wall before amplification by PCR using a ubiquitous set of primers targeting bacterial 16S rRNA (B) or <i>Pg</i> 16S rRNA (C) gene. Amplification products were separated by electrophoresis in a 1% agarose gel.</p

<i>P. gingivalis</i> (<i>Pg</i>) promotes NET formation.

<p>Freshly isolated human neutrophils were plated on Lab-tek® chamber slides and then stimulated or not by f-MLP (100 nM), a bacterial peptide used as a positive control, or by <i>Pg</i> (1.10⁷ CFU) for 2 hours at 37°C. (A) Immunofluorescence detection of histone H1 (red) and citrullinated histone H4 (green) was performed without permeabilization. (B) Cell-free DNA (cf-DNA) concentration was determined in the culture medium of neutrophils stimulated or not either with f-MLP (100 nM) or increasing concentrations of Pg. *p<0.05; **p<0.01 <i>vs</i> ctl (Mann-Whitney analysis). The trapping of Pg (red) by externalized nucleosomes (histone H1, green) was visualized by epifluorescence (C) and confocal microscopy (D). The bottom panel represents a virtual section constructed according to the Z axis, confirming the intracellular presence of <i>Pg</i> subsequent to phagocytosis by neutrophils.</p

Cell-free DNA (Cf-DNA) and MPO-DNA complexes are increased in rats infected with <i>P. gingivalis</i> (<i>Pg</i>).

<p>(A) Concentration of cf-DNA released by the AAA segment (thrombus + wall) of saline- or <i>Pg</i>-infected rats or by the thoracic aorta (T. Ao) from <i>Pg</i>-infected rats. Cf-DNA was also quantified in plasma. (B) Quantification of MPO-DNA complexes in conditioned medium or plasma of saline-or <i>Pg</i>-injected rats. **p<0.01; ***p<0.0001 (Mann-Whitney analysis). The correlation between AAA diameter and Cf-DNA concentration was determined by the Least Squares method.</p

Chronic <i>P gingivalis</i> (<i>Pg</i>) infection fostered AAA development in rats.

<p>(A) Experimental AAA was induced by implanting a segment of a SDS-decellularized guinea pig aorta in the rat aorta. Rats were or not infected weekly with <i>Pg</i> (1.10⁷ CFU/500 µL/rat, for 4 weeks). (B) At the end of treatment, rats were anaesthetized for blood sampling and sacrified after measuring AAA diameter. Results (n = 10) are presented as box plots in which median is shown, **p<0.01 (Mann-Whitney Analysis).</p

MPO-DNA complexes in conditioned medium and in plasma of human AAA.

<p>MPO-DNA complexes released by the intra-luminal thrombus (ILT) of AAA and by the arterial wall of control aorta and AAA were quantified in the conditioned media (A) as well as in plasma (B). A sandwich ELISA was used, consisting of an anti-human MPO for immunocapture and a peroxidase-conjugated anti-DNA antibody for detection. Results are presented as box plots in which the median is shown. *p<0.05, **p<0.01; ***p<0.0001 (Mann-Whitney analysis).</p