Antimicrobial susceptibility and virulence of Enterococcus spp. isolated from periodontitis-associated subgingival biofilm.

BACKGROUND: This study evaluated the prevalence, virulence and antimicrobial susceptibility of enterococci isolated from the subgingival microbiota of patients with different periodontal status. METHODS: Subgingival biofilm was obtained from individuals with periodontal health (PH) (n = 139), gingivitis (n = 103), and periodontitis (n = 305) and cultivated on selective media. Isolated strains were identified by mass spectrometry. Antimicrobial sensitivity was determined by disk diffusion, virulence genes by polymerase chain reaction, and the subgingival microbiota by checkerboard. Differences among groups were assessed by Kruskal-Wallis, Mann-Whitney, and Chi-square tests. RESULTS: Enterococcus spp. were isolated from 7.4% of all samples; 53.7% were Enterococcus faecalis. They were more prevalent in periodontitis (9.8%) and gingivitis (7.8%) than PH (2.2%; P <0.05), but no differences among stages of disease severity were observed. High rates of low susceptibility/resistance (>64%) to at least one antimicrobial were observed. Predominant virulence factors included ace (64.3%), asa (39.3%), and esp (35.7%). Fusobacterium nucleatum was prevalent in the subgingival microbiota of enterococci+ individuals, whereas Dialister pneumosintes was found in low frequency in patients with bopD+ enterococci. Oral streptococci were prevalent (>70%) in patients carrying enterococci susceptible to doxycycline (P <0.05), usually bopD- and esp- (P <0.01). CONCLUSIONS: E. faecalis is increased in periodontitis-associated biofilm. Oral enterococci carry virulence genes and express resistance to some antibiotics commonly used in dentistry, such as ciprofloxacin and erythromycin. Specific subgingival taxa are associated with oral enterococci, suggesting they may interact with species of the dysbiotic periodontitis biofilm, constituting a potential source of factors to tissue destruction, antibiotic resistance dissemination, and poor response to periodontal therapy.

Periodontal status, vascular reactivity, and platelet aggregation changes in rats submitted to hypercholesterolemic diet and periodontitis.

BACKGROUND AND OBJECTIVES: Periodontitis can corroborate with development and progression of atherosclerosis and a possible bidirectional interaction between both pathologies has been hypothesized. The aim of this work was to study the interactions between diet-induced hypercholesterolemia and ligature-induced periodontitis in Wistar rats submitted to both conditions. MATERIAL AND METHODS: Animals were divided into four experimental groups: C (control: standard diet without periodontitis), Perio (periodontitis plus standard diet), HC (high cholesterol diet without periodontitis), and HC + Perio (high cholesterol diet plus periodontitis). The diets were offered for 45 days and a silk ligature was applied in the lower first molars of Perio and HC-Perio animals on day 34 and maintained for 11 days until euthanasia. The mandibles were excised, and alveolar bone loss was determined by macroscopic and micro-tomographic (µ-CT) imaging. Blood samples were obtained, and platelet aggregation was induced in plasma rich in platelets by adenosine diphosphate (ADP) and collagen. Endothelium-dependent vascular reactivity and protein expression of endothelial (eNOS), phosphorylated endothelial (peNOS), and inducible (iNOS) nitric oxide synthases were evaluated in aorta samples. RESULTS: The HC diet combined with periodontitis (HC + Perio group) was associated with an increased alveolar bone loss, when compared to the other groups. Both in Perio and HC groups, platelet aggregation induced by ADP or collagen was increased, while maximum aortic relaxation induced by acetylcholine was decreased. Periodontitis or HC diet alone decreased the expression of peNOS and HC diet increased the expression of iNOS. In contrast, no additive or synergistic effects were found in vascular reactivity or in platelet aggregation when the two conditions were associated (HC + Perio group). CONCLUSION: Hypercholesterolemia accelerated the process of bone loss induced by periodontitis while a high cholesterol diet or periodontitis individually increased platelet aggregation and vascular reactivity in rats without additive or synergistic effects, when associated.