scholarly journals The Influence of Oral Bacteria on Epithelial Cell MigrationIn Vitro

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Alexa M. G. A. Laheij ◽  
Johannes J. de Soet ◽  
Enno C. I. Veerman ◽  
Jan G. M. Bolscher ◽  
Cor van Loveren
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Oral Bacteria ◽  
Bacterial Cells ◽  
Oral Epithelial Cells ◽  
Prevotella Nigrescens ◽  
Hematopoetic Stem Cell ◽  
Oral Epithelial ◽  
Hematopoetic Stem Cell Transplantation

Oral ulcerations often arise as a side effect from chemo- and radiation therapy. In a previous clinical study,Porphyromonas gingivaliswas identified as a positive predictor for oral ulcerations after hematopoetic stem cell transplantation, possibly incriminatingP. gingivalisin delayed healing of the ulcerations. Therefore, it was tested whetherP. gingivalisand its secreted products could inhibit the migration of oral epithelial cells in anin vitroscratch assay. To compare, the oral bacteriaPrevotella nigrescens,Prevotella intermedia,Tannerella forsythia, andStreptococcus mitiswere included. A standardized scratch was made in a confluent layer of human oral epithelial cells. The epithelial cells were challenged with bacterial cells and with medium containing secretions of these bacteria. Closure of the scratch was measured after 17 h using a phase contrast microscope.P. gingivalis,P. nigrescens, and secretions ofP. gingivalisstrongly inhibited cell migration. A challenge with 1000 heat-killed bacteria versus 1 epithelial cell resulted in a relative closure of the scratch of 25% forP. gingivalisand 20% forP. nigrescens. Weaker inhibitory effects were found for the other bacteria. The results confirmed our hypothesis that the oral bacteria may be involved in delayed wound healing.

scholarly journals Invasion of Human Oral Epithelial Cells byPrevotella intermedia

1998 ◽  
Vol 66 (12) ◽  
pp. 6054-6057 ◽  
Author(s):  
Brian R. Dorn ◽  
K.-P. Leung ◽  
Ann Progulske-Fox
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Clinical Isolate ◽  
Oral Bacteria ◽  
Epithelial Cell Line ◽  
Oral Epithelial Cells ◽  
Oral Epithelial Cell ◽  
Type C ◽  
Oral Epithelial

ABSTRACT Invasion of oral epithelial cells by pathogenic oral bacteria may represent an important virulence factor in the progression of periodontal disease. Here we report that a clinical isolate ofPrevotella intermedia, strain 17, was found to invade a human oral epithelial cell line (KB), whereas P. intermedia 27, another clinical isolate, and P. intermedia 25611, the type strain, were not found to invade the cell line. Invasion was quantified by the recovery of viable bacteria following a standard antibiotic protection assay and observed by electron microscopy. Cytochalasin D, cycloheximide, monodansylcadaverine, and low temperature (4°C) inhibited the internalization of P. intermedia 17. Antibodies raised against P. intermedia type C fimbriae and against whole cells inhibited invasion, but the anti-type-C-fimbria antibody inhibited invasion to a greater extent than the anti-whole-cell antibody. This work provides evidence that at least one strain ofP. intermedia can invade an oral epithelial cell line and that the type C fimbriae and a cytoskeletal rearrangement are required for this invasion.

scholarly journals In Vitro Studies on Erythrosine-Based Photodynamic Therapy of Malignant and Pre-Malignant Oral Epithelial Cells

PLoS ONE ◽  
2012 ◽  
Vol 7 (4) ◽  
pp. e34475 ◽  
Author(s):  
Abhishek D. Garg ◽  
Muthiah Bose ◽  
Mohammed I. Ahmed ◽  
William A. Bonass ◽  
Simon R. Wood
Keyword(s):  
Photodynamic Therapy ◽  
Epithelial Cells ◽  
In Vitro Studies ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

scholarly journals Interactions between Periodontal Bacteria and Human Oral Epithelial Cells: Fusobacterium nucleatum Adheres to and Invades Epithelial Cells

2000 ◽  
Vol 68 (6) ◽  
pp. 3140-3146 ◽  
Author(s):  
Yiping W. Han ◽  
Wenyuan Shi ◽  
George T.-J. Huang ◽  
Susan Kinder Haake ◽  
No-Hee Park ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Periodontal Diseases ◽  
Fusobacterium Nucleatum ◽  
Oral Bacteria ◽  
Metabolic Inhibitors ◽  
Kb Cells ◽  
Gingival Epithelial Cells ◽  
Oral Epithelial

ABSTRACT Bacteria are causative agents of periodontal diseases. Interactions between oral bacteria and gingival epithelial cells are essential aspects of periodontal infections. Using an in vitro tissue culture model, a selected group of gram-negative anaerobic bacteria frequently associated with periodontal diseases, includingBacteroides forsythus, Campylobacter curvus,Eikenella corrodens, Fusobacterium nucleatum,Porphyromonas gingivalis, and Prevotella intermedia, were examined for their ability to adhere to and invade primary cultures of human gingival epithelial cells (HGEC). The effects of these bacteria on the production of interleukin-8 (IL-8), a proinflammatory chemokine, were also measured. These studies provided an initial demonstration that F. nucleatum adhered to and invaded HGEC and that this was accompanied by high levels of IL-8 secretion from the epithelial cells. The attachment and invasion characteristics of F. nucleatumwere also tested using KB cells, an oral epithelial cell line. The invasion was verified by transmission electron microscopy and with metabolic inhibitors. Invasion appeared to occur via a “zipping” mechanism and required the involvement of actins, microtubules, signal transduction, protein synthesis, and energy metabolism of the epithelial cell, as well as protein synthesis by F. nucleatum. A spontaneous mutant, lam, of F. nucleatum, isolated as defective in autoagglutination, was unable to attach to or invade HGEC or KB cells, further indicating the requirement of bacterial components in these processes. Sugar inhibition assays indicated that lectin-like interactions were involved in the attachment of F. nucleatum to KB cells. Investigation of these new virulence phenotypes should improve our understanding of the role of F. nucleatum in periodontal infections.

scholarly journals The Aryl Hydrocarbon Receptor Governs Epithelial Cell Invasion during Oropharyngeal Candidiasis

mBio ◽  
2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Norma V. Solis ◽  
Marc Swidergall ◽  
Vincent M. Bruno ◽  
Sarah L. Gaffen ◽  
Scott G. Filler
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Signaling Pathways ◽  
Oral Epithelium ◽  
New Approach ◽  
Oral Epithelial Cells ◽  
Aryl Hydrocarbon ◽  
Molecule Inhibitor ◽  
Oral Epithelial ◽  
Cell Signaling Pathways

ABSTRACT Oropharyngeal candidiasis (OPC), caused predominantly by Candida albicans, is a prevalent infection in patients with advanced AIDS, defects in Th17 immunity, and head and neck cancer. A characteristic feature of OPC is fungal invasion of the oral epithelial cells. One mechanism by which C. albicans hyphae can invade oral epithelial cells is by expressing the Als3 and Ssa1 invasins that interact with the epidermal growth factor receptor (EGFR) on epithelial cells and stimulate endocytosis of the organism. However, the signaling pathways that function downstream of EGFR and mediate C. albicans endocytosis are poorly defined. Here, we report that C. albicans infection activates the aryl hydrocarbon receptor (AhR), leading to activation of Src family kinases (SFKs), which in turn phosphorylate EGFR and induce endocytosis of the fungus. Furthermore, treatment of oral epithelial cells with interferon gamma inhibits fungal endocytosis by inducing the synthesis of kynurenines, which cause prolonged activation of AhR and SFKs, thereby interfering with C. albicans-induced EGFR signaling. Treatment of both immunosuppressed and immunocompetent mice with an AhR inhibitor decreases phosphorylation of SFKs and EGFR in the oral mucosa, reduces fungal invasion, and lessens the severity of OPC. Thus, our data indicate that AhR plays a central role in governing the pathogenic interactions of C. albicans with oral epithelial cells during OPC and suggest that this receptor is a potential therapeutic target. IMPORTANCE OPC is caused predominantly by the fungus C. albicans, which can invade the oral epithelium by several mechanisms. One of these mechanisms is induced endocytosis, which is stimulated when fungal invasins bind to epithelial cell receptors such as EGFR. Receptor binding causes rearrangement of epithelial cell microfilaments, leading to the formation of pseudopods that engulf the fungus and pull it into the epithelial cell. We discovered AhR acts via SFKs to phosphorylate EGFR and induce the endocytosis of C. albicans. Our finding that a small molecule inhibitor of AhR ameliorates OPC in mice suggests that a strategy of targeting host cell signaling pathways that govern epithelial cell endocytosis of C. albicans holds promise as a new approach to preventing or treating OPC. IMPORTANCE OPC is caused predominantly by the fungus C. albicans, which can invade the oral epithelium by several mechanisms. One of these mechanisms is induced endocytosis, which is stimulated when fungal invasins bind to epithelial cell receptors such as EGFR. Receptor binding causes rearrangement of epithelial cell microfilaments, leading to the formation of pseudopods that engulf the fungus and pull it into the epithelial cell. We discovered AhR acts via SFKs to phosphorylate EGFR and induce the endocytosis of C. albicans. Our finding that a small molecule inhibitor of AhR ameliorates OPC in mice suggests that a strategy of targeting host cell signaling pathways that govern epithelial cell endocytosis of C. albicans holds promise as a new approach to preventing or treating OPC.

scholarly journals Tannerella forsythia invasion in oral epithelial cells requires phosphoinositide 3-kinase activation and clathrin-mediated endocytosis

Microbiology ◽  
2011 ◽  
Vol 157 (8) ◽  
pp. 2382-2391 ◽  
Author(s):  
Elina Mishima ◽  
Ashu Sharma
Keyword(s):  
Epithelial Cells ◽  
Rho Gtpases ◽  
Membrane Microdomains ◽  
Small Interfering Rnas ◽  
Tannerella Forsythia ◽  
Lipid Kinase ◽  
Signalling Molecules ◽  
Oral Epithelial Cells ◽  
Oral Epithelial

Tannerella forsythia, a Gram-negative anaerobe implicated in periodontitis, has been detected within human buccal epithelial cells and shown to invade oral epithelial cells in vitro. We have previously shown that this bacterium triggers host tyrosine kinase-dependent phosphorylation and actin-dependent cytoskeleton reorganization for invasion. On the bacterial side, the leucine-rich repeat cell-surface BspA protein is important for entry. The present study was undertaken to identify host signalling molecules during T. forsythia entry into human oral and cervical epithelial cells. Specifically, the roles of phosphatidylinositol 3-kinase (PI3K), Rho-family GTPases, cholesterol-rich membrane microdomains and the endocytic protein clathrin were investigated. For this purpose, cell lines were pretreated with chemical inhibitors or small interfering RNAs (siRNAs) that target PI3Ks, Rho GTPases, clathrin and cholesterol (a critical component of ‘lipid rafts’), and the resulting effects on T. forsythia uptake were determined. Our studies revealed that T. forsythia entry is dependent on host PI3K signalling, and that purified BspA protein causes activation of this lipid kinase. Bacterial entry also requires the cooperation of host Rac1 GTPase. Finally, our findings indicate an important role for clathrin and cholesterol-rich lipid microdomains in the internalization process

scholarly journals Calprotectin Expression In Vitro by Oral Epithelial Cells Confers Resistance to Infection by Porphyromonas gingivalis

2001 ◽  
Vol 69 (7) ◽  
pp. 4242-4247 ◽  
Author(s):  
K. Nisapakultorn ◽  
K. F. Ross ◽  
M. C. Herzberg
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Porphyromonas Gingivalis ◽  
Calcium Binding ◽  
Periodontal Diseases ◽  
Epithelial Cell Line ◽  
Transfected Cells ◽  
Cell Rounding ◽  
Oral Epithelial

ABSTRACT Calprotectin, an S100 calcium-binding protein with broad-spectrum antimicrobial activity in vitro, is expressed in neutrophils, monocytes, and gingival keratinocytes. In periodontitis, calprotectin appears upregulated and is detected at higher levels in gingival crevicular fluid and tissue specimens. How calprotectin contributes to the pathogenesis of periodontal diseases is unknown. To isolate the effects of calprotectin, a calprotectin-negative oral epithelial cell line was transfected with calprotectin genes to enable expression.Porphyromonas gingivalis was permitted to bind and invade transfected cells expressing calprotectin and sham transfectants. Rates of invasion into both cell lines were compared using the antibiotic protection assay. Transfected cells expressing calprotectin showed 40 to 50% fewer internalized P. gingivalis than sham transfectants. Similarly, binding to calprotectin expressing cells was reduced approximately twofold at all time points (15, 30, 45, and 60 min) as estimated by immunofluorescence analysis. Independent of invasion, however, prolonged exposure to P. gingivalisinduced epithelial cell rounding and detachment from the substratum. These morphological changes were delayed, however, in cells expressing calprotectin. Using P. gingivalis protease-deficient mutants, we found that Arg-gingipain and Lys-gingipain contributed to epithelial cell rounding and detachment. In conclusion, expression of calprotectin appears to protect epithelial cells in culture against binding and invasion by P. gingivalis. In addition, cells expressing calprotectin are more resistant to detachment mediated by Arg-gingipain and Lys-gingipain. In periodontal disease, calprotectin may augment both the barrier protection and innate immune functions of the gingival epithelium to promote resistance to P. gingivalis infection.

scholarly journals An In Vitro Model of Ciprofloxacin and Minocycline Transport by Oral Epithelial Cells

2002 ◽  
Vol 73 (11) ◽  
pp. 1267-1272 ◽  
Author(s):  
James J. Brayton ◽  
Qing Yang ◽  
Robin J. Nakkula ◽  
John D. Walters
Keyword(s):  
Epithelial Cells ◽  
In Vitro Model ◽  
Oral Epithelial Cells ◽  
Vitro Model ◽  
Oral Epithelial

scholarly journals Potential Role for a Carbohydrate Moiety in Anti-Candida Activity of Human Oral Epithelial Cells

2001 ◽  
Vol 69 (11) ◽  
pp. 7091-7099 ◽  
Author(s):  
Chad Steele ◽  
Janet Leigh ◽  
Rolf Swoboda ◽  
Hatice Ozenci ◽  
Paul L. Fidel
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Oral Mucosa ◽  
Host Defense ◽  
Defense Mechanisms ◽  
Carbohydrate Moiety ◽  
Sulfated Polysaccharides ◽  
Oral Epithelial Cells ◽  
Oral Epithelial Cell ◽  
Oral Epithelial

ABSTRACT Candida albicans is both a commensal and a pathogen at the oral mucosa. Although an intricate network of host defense mechanisms are expected for protection against oropharyngeal candidiasis, anti-Candida host defense mechanisms at the oral mucosa are poorly understood. Our laboratory recently showed that primary epithelial cells from human oral mucosa, as well as an oral epithelial cell line, inhibit the growth of blastoconidia and/or hyphal phases of several Candida species in vitro with a requirement for cell contact and with no demonstrable role for soluble factors. In the present study, we show that oral epithelial cell-mediated anti-Candida activity is resistant to gamma-irradiation and is not mediated by phagocytosis, nitric oxide, hydrogen peroxide, and superoxide oxidative inhibitory pathways or by nonoxidative components such as soluble defensin and calprotectin peptides. In contrast, epithelial cell-mediated anti-Candida activity was sensitive to heat, paraformaldehyde fixation, and detergents, but these treatments were accompanied by a significant loss in epithelial cell viability. Treatments that removed existing membrane protein or lipid moieties in the presence or absence of protein synthesis inhibitors had no effect on epithelial cell inhibitory activity. In contrast, the epithelial cell-mediated anti-Candida activity was abrogated after treatment of the epithelial cells with periodic acid, suggesting a role for carbohydrates. Adherence of C. albicans to oral epithelial cells was unaffected, indicating that the carbohydrate moiety is exclusively associated with the growth inhibition activity. Subsequent studies that evaluated specific membrane carbohydrate moieties, however, showed no role for sulfated polysaccharides, sialic acid residues, or glucose- and mannose-containing carbohydrates. These results suggest that oral epithelial cell-mediated anti-Candida activity occurs exclusively with viable epithelial cells through contact with C. albicans by an as-yet-undefined carbohydrate moiety.

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