Relationships between subgingival microbiota and GCF biomarkers in generalized aggressive periodontitis

Abstract

Aim-To examine relationships between subgingival biofilm composition and levels of gingival crevicular fluid (GCF) cytokines in periodontal health and generalized aggressive periodontitis (GAP).Materials and methods-Periodontal parameters were measured in 25 periodontally healthy and 31 GAP subjects. Subgingival plaque and GCF samples were obtained from 14 sites from each subject. 40 subgingival taxa were quantified using checkerboard DNA-DNA hybridization and the concentrations of 8 GCF cytokines measured using Luminex. Cluster analysis was used to define sites with similar subgingival microbiotas in each clinical group. Significance of differences in clinical, microbiological and immunological parameters among clusters was determined using the Kruskal-Wallis test.Results-GAP subjects had statistically significantly higher GCF levels of interleukin-1β (IL-1β) (p<0.001), granulocyte-macrophage colony-stimulating factor (GM-CSF) (p<0.01), and IL-1β/IL-10 ratio (p<0.001) and higher proportions of Red and Orange complex species than periodontally healthy subjects. There were no statistically significant differences in the mean proportion of cytokines among clusters in the periodontally healthy subjects, while the ratio IL-1β/ IL-10 (p<0.05) differed significantly among clusters in the aggressive periodontitis group.Conclusions-Different subgingival biofilm profiles are associated with distinct patterns of GCF cytokine expression. Aggressive periodontitis subjects were characterized by a higher IL-1β/ IL-10 ratio than periodontally healthy subjects, suggesting an imbalance between pro-and antiinflammatory cytokines in aggressive periodontitis. KeywordsGeneralized aggressive periodontitis; microbiota; gingival crevicular fluid; biomarkers Clinical RelevanceScientific rationale for study: Periodontal diseases result from interactions between specific subgingival microbial species and the susceptible host. However, very little is known regarding the impact of the subgingival biofilm composition on the secretion of biomarkers by the adjacent periodontal tissues.

Interactions between the Aggregatibacter actinomycetemcomitans secretin HofQ and host cytokines indicate a link between natural competence and interleukin-8 uptake

“…Twenty-five nanograms of Delfia Eu-N1 anti-6xHis antibody (Perkin Elmer, AD0108) in 50 μl of Delfia® Assay Buffer was incubated in wells at RT for 1 h, after which the wells were washed as described above. A 5-min incubation with DELFIA® Enhancement Solution (Perkin Elmer, 4001–0010) was followed by measuring time-resolved fluorescence using a Victor [3] multilabel plate reader (Perkin Elmer). Each sample was prepared in triplicate, and three independent experiments were performed.…”

“…Plasma/serum was collected from 3 ml of (EDTA-coagulated) whole blood after centrifugation (1000 × g, 10 min, RT), and leukocytes were freshly isolated from 1 ml of (EDTA-coagulated) whole blood following a protocol by Lilius and Nuutila [51]. Measurement of chemiluminescence (every 2 min for 200 min; Victor [3] multilabel plate reader (Perkin Elmer) or Hidex Sense microplate reader (Hidex)) was started immediately after leukocytes were added to the reaction. When assessing the induction of ROS production in leukocytes by the A. actinomycetemcomitans D7S wild-type and ΔhofQ mutant strains, the plasma used was obtained from the same healthy individual, while the leukocytes were from seven different healthy individuals.…”

“…Aggregatibacter actinomycetemcomitans is a gram-negative, opportunistic, oral pathogen that is associated with both aggressive and chronic forms of periodontitis [1–3]. In periodontitis, an intensified host inflammatory response against subgingival biofilms, consisting primarily of inflammophilic gram-negative species, causes the destruction of gingival tissue and alveolar bone (for reviews see refs.…”

Clinical and Microbiological Characterization of Patients with Generalized Aggressive Periodontitis

“…The composition of the subgingival microbiota among subjects with GAgP usually differs considerably (Teles et al, 2010). In recognized disease and in exacting, after the extent of periodontal lesions, the microbiota might turn into much more complex and soon may resemble that of generalized chronic periodontitis with microorganisms of the red complex apparently dominating the subgingival ecosystems (Könönen & Müller).…”

IL-1 Receptor Type II Production Is Upregulated by IL-4 and IL-13, and Downregulated by IFN-&lt;i&gt;γ&lt;/i&gt; in Mouse Gingival Epithelial Cells

“…Subgingival biofilm plays a major role in the pathogenesis of periodontal disease by stimulating an immune response that can lead to periodontal breakdown [1] [2]. In addition to genetic and acquired risk factors that can modify host response, the susceptibility to periodontal disease, as well as its severity and progression is influenced by environmental factors [3]- [5].…”