Quantitative detection of Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa in human oral epithelial cells from subjects with periodontitis and periodontal health

Abstract

However, non-oral pathogens internalized in oral epithelial cells and their relationship with periodontal status are poorly understood. The purpose of this study was to detect opportunistic species within buccal and gingival crevice epithelial cells collected from subjects with periodontitis or individuals with good periodontal health, and to associate their prevalence with periodontal clinical status. Quantitative detection of total bacteria and Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis in oral epithelial cells was determined by quantitative real-time PCR using universal and species-specific primer sets. Intracellular bacteria were visualized by confocal microscopy and fluorescence in situ hybridization. Overall, 33 % of cell samples from patients with periodontitis contained at least one opportunistic species, compared with 15 % of samples from healthy individuals. E. faecalis was the most prevalent species found in oral epithelial cells (detected in 20.6 % of patients with periodontitis, P50.03 versus healthy individuals) and was detected only in cells from patients with periodontitis. Quantitative real-time PCR showed that high levels of P. aeruginosa and S. aureus were present in both the periodontitis and healthy groups. However, the proportion of these species was significantly higher in epithelial cells of subjects with periodontitis compared with healthy individuals (P50.016 for P. aeruginosa and P50.047 for S. aureus). Although E. faecalis and P. aeruginosa were detected in 57 % and 50 % of patients, respectively, with probing depth and clinical attachment level ¢6 mm, no correlation was found with age, sex, bleeding on probing or the presence of supragingival biofilm. The prevalence of these pathogens in epithelial cells is correlated with the state of periodontal disease. INTRODUCTIONThe oral cavity harbours a complex microbiota that is commonly related to oral health. Nonetheless, it is common knowledge that only some microbial consortia are associated with carious lesions and periodontal disease (Aas et al., 2005;Socransky & Haffajee, 2005). On the other hand, classically pathogenic micro-organisms can find favourable environmental conditions within and occupy specific niches of the oral cavity (Fourrier et al., 1998). Bacterial species commonly associated with nosocomial infections and multiresistance to antimicrobials, such as Pseudomonas aeruginosa, Staphylococcus aureus, Acinetobacter baumannii, Enterococcus faecalis and the Enterobacteriaceae family, have been detected in high proportions and levels in the subgingival biofilm of individuals with periodontitis (Slots et al., 1990;Colombo et al., 2002) and in the endodontic canal (Brito et al., 2007). Furthermore, there seems to be a strong association between the presence of these pathogens in dental plaque and respiratory infections in hospitalized patients (El-Solh et al., 2004;Didilescu et al., 2005;Bahrani-Mougeot et al., 2007).Abbreviations: BOP, bleeding on probing; CAL, clinical attachment level; FISH, fluo...

Mucosal Microbiome in Patients with Recurrent Aphthous Stomatitis

“…Total genera from the order Bacteroidales comprising several Gram-negative microorganisms of the normal colonic microbiota as well as oral opportunistic pathogens (Colombo et al 2013) predominated in healthy sites from RAS patients over HCs. Porphyromonadaceae comprising species specifically associated with periodontal disease (Mysak et al 2014) and Veillonellaceae for which diverse roles in oral health and disease are reported predominated only in ulcerated sites over HC.…”

Antimicrobial and anti-inflammatory potential therapy for opportunistic microorganisms

“…The most represented opportunistic microorganisms include Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans [2,3]. Unfortunately, due to the overuse and misuse of antibiotics, most of these organisms become drug resistant, leading to difficulties in curing related infectious diseases and thus reducing therapeutic options [3,4].…”

Relationship between invasion of theperiodontiumby periodontal pathogens and periodontal disease: a systematic review

“…One study was in Japanese and was excluded. 36 Twenty-two studies were subsequently excluded for specific reasons: 3 studies 34,35,37 had a study protocol not related to the objectives of this systematic review, 8 studies 14,[25][26][27]30,39,40,42 had absence of control group (non-diseased samples), one study 38 only presented disease samples from a single patient, 7 studies 28,29,[31][32][33]41,44 hadn't clearly defined inclusion and/or exclusion criteria, and finally 3 studies 43,45,46 were excluded because they only addressed opportunistic pathogens. Additional hand searching of the reference lists of the selected papers didn't retrieve any additional studies.…”