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

2013 ◽  
Vol 62 (10) ◽  
pp. 1592-1600 ◽  
Author(s):  
Andrea V. Colombo ◽  
Graziela M. Barbosa ◽  
Daniela Higashi ◽  
Giorgio di Micheli ◽  
Paulo H. Rodrigues ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Epithelial Cells ◽  
Periodontal Disease ◽  
Quantitative Detection ◽  
Healthy Individuals ◽  
Quantitative Real Time Pcr ◽  
Periodontal Health ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

Epithelial cells in oral cavities can be considered reservoirs for a variety of bacterial species. A polymicrobial intracellular flora associated with periodontal disease has been demonstrated in buccal cells. Important aetiological agents of systemic and nosocomial infections have been detected in the microbiota of subgingival biofilm, especially in individuals with periodontal disease. 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, P = 0.03 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 (P = 0.016 for P. aeruginosa and P = 0.047 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.

scholarly journals Growth and adherence of Staphylococcus aureus were enhanced through the PGE2 produced by the activated COX-2/PGE2 pathway of infected oral epithelial cells

PLoS ONE ◽  
2017 ◽  
Vol 12 (5) ◽  
pp. e0177166 ◽  
Author(s):  
Yuxia Wang ◽  
Biao Ren ◽  
Xuedong Zhou ◽  
Shiyu Liu ◽  
Yujie Zhou ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Epithelial Cells ◽  
Oral Epithelial Cells ◽  
Cox 2 ◽  
Oral Epithelial

scholarly journals Arginine-Specific Protease fromPorphyromonas gingivalis Activates Protease-Activated Receptors on Human Oral Epithelial Cells and Induces Interleukin-6 Secretion

2001 ◽  
Vol 69 (8) ◽  
pp. 5121-5130 ◽  
Author(s):  
Afrodite Lourbakos ◽  
Jan Potempa ◽  
James Travis ◽  
Michael R. D'Andrea ◽  
Patricia Andrade-Gordon ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Periodontal Disease ◽  
Intracellular Calcium ◽  
Cell Line ◽  
Interleukin 6 ◽  
Epithelial Cell Line ◽  
Protease Activated Receptors ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

ABSTRACT Periodontitis is a chronic inflammatory disease affecting oral tissues. Oral epithelial cells represent the primary barrier against bacteria causing the disease. We examined the responses of such cells to an arginine-specific cysteine proteinase (RgpB) produced by a causative agent of periodontal disease, Porphyromonas gingivalis. This protease caused an intracellular calcium transient in an oral epithelial cell line (KB), which was dependent on its enzymatic activity. Since protease-activated receptors (PARs) might mediate such signaling, reverse transcription-PCR was used to characterize the range of these receptors expressed in the KB cells. The cells were found to express PAR-1, PAR-2, and PAR-3, but not PAR-4. In immunohistochemical studies, human gingival epithelial cells were found to express PAR-1, PAR-2, and PAR-3 on their surface, but not PAR-4, indicating that the cell line was an effective model for the in vivo situation. PAR-1 and PAR-2 expression was confirmed in intracellular calcium mobilization assays by treatment of the cells with the relevant receptor agonist peptides. Desensitization experiments strongly indicated that signaling of the effects of RgpB was occurring through PAR-1 and PAR-2. Studies with cells individually transfected with each of these two receptors confirmed that they were both activated by RgpB. Finally, it was shown that, in the oral epithelial cell line, PAR activation by the bacterial protease-stimulated secretion of interleukin-6. This induction of a powerful proinflammatory cytokine suggests a mechanism whereby cysteine proteases from P. gingivalis might mediate inflammatory events associated with periodontal disease on first contact with a primary barrier of cells.

scholarly journals Effects of Short-Chain Fatty Acids on Human Oral Epithelial Cells and the Potential Impact on Periodontal Disease: A Systematic Review of In Vitro Studies

2020 ◽  
Vol 21 (14) ◽  
pp. 4895
Author(s):  
Gabriel Leonardo Magrin ◽  
Franz Josef Strauss ◽  
Cesar Augusto Magalhães Benfatti ◽  
Lucianne Cople Maia ◽  
Reinhard Gruber
Keyword(s):  
Fatty Acids ◽  
Epithelial Cells ◽  
Periodontal Disease ◽  
Short Chain Fatty Acids ◽  
Risk Of Bias ◽  
In Vitro Studies ◽  
Short Chain ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

Short-chain fatty acids (SCFA), bacterial metabolites released from dental biofilm, are supposed to target the oral epithelium. There is, however, no consensus on how SCFA affect the oral epithelial cells. The objective of the present study was to systematically review the available in vitro evidence of the impact of SCFA on human oral epithelial cells in the context of periodontal disease. A comprehensive electronic search using five databases along with a grey literature search was performed. In vitro studies that evaluated the effects of SCFA on human oral epithelial cells were eligible for inclusion. Risk of bias was assessed by the University of Bristol’s tool for assessing risk of bias in cell culture studies. Certainty in cumulative evidence was evaluated using GRADE criteria (grading of recommendations assessment, development, and evaluation). Of 3591 records identified, 10 were eligible for inclusion. A meta-analysis was not possible due to the heterogeneity between the studies. The risk of bias across the studies was considered “serious” due to the presence of methodological biases. Despite these limitations, this review showed that SCFA negatively affect the viability of oral epithelial cells by activating a series of cellular events that includes apoptosis, autophagy, and pyroptosis. SCFA impair the integrity and presumably the transmigration of leucocytes through the epithelial layer by changing junctional and adhesion protein expression, respectively. SCFA also affect the expression of chemokines and cytokines in oral epithelial cells. Future research needs to identify the underlying signaling cascades and to translate the in vitro findings into preclinical models.

scholarly journals Age and AgNor- A Morphometric study

2020 ◽  
Vol 1 (1) ◽  
Author(s):  
Manpreet Kaur ◽  
Smita Naik ◽  
Sachin Jindal
Keyword(s):  
Epithelial Cells ◽  
Buccal Mucosa ◽  
Age Groups ◽  
Morphometric Study ◽  
Healthy Individuals ◽  
Oral Epithelial Cells ◽  
Glass Slides ◽  
Buccal Epithelial Cells ◽  
Oral Epithelial ◽  
Ribosome Biosynthesis

To investigate and correlate total AgNOR area/Total nucleus area (TAA/TNA ) values in buccal epithelial cells of healthy individuals in different age groups. Material and Methodology:- In present study 50 healthy individuals are included with age ranging 10 to 60 years. These are divided into 5 groups.Group1-10-20year, Group2-20- 30year, Group3-30-40year, Group4-40-50year, Group5-50-60year. Oral epithelial cells collected with the help of cyto brush from buccal mucosa. Smears were prepared on clean glass slides and fixed with 95% alcohol. Fixed slides Smears were stained with AgNOR stain. Results: Statistically significant correlations were found between mean TAA/TNA values and age p < 0.001 for linear and p < 0.0001 for polynominal regression, and between AgNOR number and age, p < 0.001 for linear. Conclusion: There is a significant correlation between age and AgNOR amount (ribosome biosynthesis rate) in buccal epithelial cells of healthy individuals. AgNORs in buccal epithelial cells may be used for detection of age.

scholarly journals Proto-Oncogenes and Cell Cycle Gene Expression in Normal and Neoplastic Oral Epithelial Cells Stimulated With Soluble Factors From Single and Dual Biofilms of Candida albicans and Staphylococcus aureus

2021 ◽  
Vol 11 ◽  
Author(s):  
María Isabel Amaya Arbeláez ◽  
Ana Carolina Alves de Paula e Silva ◽  
Geovana Navegante ◽  
Valeria Valente ◽  
Paula Aboud Barbugli ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Staphylococcus Aureus ◽  
Candida Albicans ◽  
Epithelial Cells ◽  
Cell Viability ◽  
Cell Cycle Gene ◽  
Soluble Factors ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

This study was aimed at analyzing proto-oncogenic signaling pathway activation in normal oral keratinocytes (NOK-si) and neoplastic cell lines (SCC 25 and Detroit 562) stimulated with metabolites (soluble factors) from single and dual biofilms of Candida albicans and Staphylococcus aureus. Soluble factors (SF) from early (16-h) and mature (36-h) biofilms of C. albicans and S. aureus were collected and incubated with cell cultures, which were subsequently evaluated using gene expression via RT-qPCR, cell viability via AlamarBlueTM, and flow cytometry cell cycle analysis. In general, exposure to the SF of early and mature biofilms from C. albicans and dual species caused a major reduction in NOK-si cell viability and enhanced the sub G0 phase. This led to a decrease in gene expression. However, in this cell line, SF of S. aureus biofilms upregulated the CDKN1A gene followed by the maintenance of cell viability and a significant increase in the G2/M population. For tumor cells, SCC 25 and Detroit 562, the stimuli of SF biofilms upregulated oncogenes such as hRAS and mTOR, as well as Bcl-2 and CDKN1A. SCC 25 and Detroit 562 cells could survive even after 24 h of stimuli from both SF (early and mature). This occurred without significant changes taking place in the cell cycle progression for SCC 25, and with a significant tendency to increase the G2/M phase for Detroit 562. These results point to the fact that metabolites from prevalent clinical fungal and bacterial biofilms, C. albicans and S. aureus, can disrupt the homeostasis of normal and neoplastic oral epithelial cells. This changes proto-oncogenes’ expression, specifically PI3KCA, hRAS, mTOR, BRAF, and cell cycle genes CDKN1A and Bcl-2, thus causing a disturbance in cell viability, survival, and the cell cycle profile.

scholarly journals Novel Aggregation Properties of Candida albicans Secreted Aspartyl Proteinase Sap6 Mediate Virulence in Oral Candidiasis

2015 ◽  
Vol 83 (7) ◽  
pp. 2614-2626 ◽  
Author(s):  
Rohitashw Kumar ◽  
Darpan Saraswat ◽  
Swetha Tati ◽  
Mira Edgerton
Keyword(s):  
Candida Albicans ◽  
Epithelial Cells ◽  
Oral Candidiasis ◽  
Oral Microbiome ◽  
Proteinase Activity ◽  
Adhesion Properties ◽  
Content Type ◽  
Oral Epithelial Cells ◽  
Oral Epithelial ◽  
Cell Cell

Candida albicans, a commensal fungus of the oral microbiome, causes oral candidiasis in humans with localized or systemic immune deficiencies. Secreted aspartic proteinases (Saps) are a family of 10 related proteases and are virulence factors due to their proteolytic activity, as well as their roles in adherence and colonization of host tissues. We found that mice infected sublingually withC. albicanscells overexpressing Sap6 (SAP6OE and a Δsap8strain) had thicker fungal plaques and more severe oral infection, while infection with the Δsap6strain was attenuated. These hypervirulent strains had highly aggregative colony structurein vitroand higher secreted proteinase activity; however, the levels of proteinase activity ofC. albicansSaps did not uniformly match their abilities to damage cultured oral epithelial cells (SCC-15 cells). Hyphal induction in cells overexpressing Sap6 (SAP6OE and Δsap8cells) resulted in formation of large cell-cell aggregates. These aggregates could be produced in germinated wild-type cells by addition of native or heat-inactivated Sap6. Sap6 bound only to germinated cells and increasedC. albicansadhesion to oral epithelial cells. The adhesion properties of Sap6 were lost upon deletion of its integrin-binding motif (RGD) and could be inhibited by addition of RGD peptide or anti-integrin antibodies. Thus, Sap6 (but not Sap5) has an alternative novel function in cell-cell aggregation, independent of its proteinase activity, to promote infection and virulence in oral candidiasis.

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