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 Potential Suppressive Effect of Nicotine on the Inflammatory Response in Oral Epithelial Cells: An In Vitro Study

2021 ◽  
Vol 18 (2) ◽  
pp. 483
Author(s):  
Na An ◽  
Jasmin Holl ◽  
Xuekui Wang ◽  
Marco Aoqi Rausch ◽  
Oleh Andrukhov ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Periodontal Diseases ◽  
Suppressive Effect ◽  
Cell Surface Expression ◽  
Epithelial Cell Line ◽  
Oral Epithelial Cells ◽  
Tnf Α ◽  
Oral Epithelial ◽  
The Impact

Smoking is a well-recognized risk factor for oral mucosal and periodontal diseases. Nicotine is an important component of cigarette smoke. This study aims to investigate the impact of nicotine on the viability and inflammatory mediator production of an oral epithelial cell line in the presence of various inflammatory stimuli. Oral epithelial HSC-2 cells were challenged with nicotine (10−8–10−2 M) for 24 h in the presence or absence of Porphyromonas gingivalis lipopolysaccharide (LPS, 1 µg/mL) or tumor necrosis factor (TNF)-α (10−7 M) for 24 h. The cell proliferation/viability was determined by MTT assay. Gene expression of interleukin (IL)-8, intercellular adhesion molecule (ICAM)-1, and β-defensin was assayed by qPCR. The production of IL-8 protein and cell surface expression of ICAM-1 was assessed by ELISA and flow cytometry, respectively. Proliferation/viability of HSC-2 cells was unaffected by nicotine at concentrations up to 10−3 M and inhibited at 10−2 M. Nicotine had no significant effect on the basal expression of IL-8, ICAM-1, and β-defensin. At the same time, it significantly diminished P. gingivalis LPS or the TNF-α-induced expression levels of these factors. Within the limitations of this study, the first evidence was provided in vitro that nicotine probably exerts a suppressive effect on the production of inflammatory mediators and antimicrobial peptides in human oral epithelial cells.

scholarly journals Listeria monocytogenes Infection of Bat Pipistrellus nathusii Epithelial cells Depends on the Invasion Factors InlA and InlB

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 867
Author(s):  
Olga Povolyaeva ◽  
Yaroslava Chalenko ◽  
Egor Kalinin ◽  
Olga Kolbasova ◽  
Elena Pivova ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Domestic Animals ◽  
Epithelial Cell Line ◽  
Intracellular Pathogen ◽  
Wild Type ◽  
Listeria Monocytogenes Infection ◽  
Natural Hosts

L. monocytogenes is a widespread facultative intracellular pathogen. The range of natural hosts that supporting L. monocytogenes persistence in the environment has not been fully established yet. In this study, we were interested in the potential of L. monocytogenes to infect cells of bats, which are being increasingly recognized as a reservoir for microorganisms that are pathogenic to humans and domestic animals. A stable epithelial cell line was developed from the kidneys of Pipistrellus nathusii, a small bat widely distributed across Europe. The wild-type L. monocytogenes strain EGDe infected this cell line with an invasion efficiency of 0.0078 ± 0.0009%. Once it entered bat cells, L. monocytogenes doubled within about 70 min. When L. monocytogenes lacked either of the major invasion factors, InlA and InlB, invasion efficiency decreased by a factor of 10 and 25 respectively (p < 0.000001). The obtained results suggest that bat epithelial cells are susceptible to L. monocytogenes infection and that L. monocytogenes invasion of bat cells depends on the major invasion factors InlA and InlB. These results constitute the first report on in vitro studies of L. monocytogenes infection in bats.

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 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.

Porphyromonas gingivalis FDC381 multiplies and persists within human oral epithelial cells in vitro.

1996 ◽  
Vol 64 (2) ◽  
pp. 660-664 ◽  
Author(s):  
P N Madianos ◽  
P N Papapanou ◽  
U Nannmark ◽  
G Dahlén ◽  
J Sandros
Keyword(s):  
Epithelial Cells ◽  
Porphyromonas Gingivalis ◽  
Oral Epithelial Cells ◽  
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 Establishment and Characterization of a Lactating Caprine Mammary Gland Luminal Epithelial Cell Line

2019 ◽  
Vol 11 (4) ◽  
pp. 36
Author(s):  
Mahipal Singh ◽  
Benjamin Hortman ◽  
Venkata Degala ◽  
Xiaoling Ma
Keyword(s):  
Mammary Gland ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Milk Production ◽  
Cell Cultures ◽  
Epithelial Cell Line ◽  
Luminal Epithelial Cell ◽  
Saanen Goat

Mammary gland is a defining characteristic of mammalian species which produces nutritious milk and plays a major role in the development of newborns. The gland contains a series of ducts and crevices leading back to alveoli, which contain milk producing cells called luminal epithelial cells. These cells, if cultured in-vitro, can be utilized to explore the metabolic processes occurring during milk production. The knowledge thus gained can be used to manipulate the system to enhance milk production and/or modify its composition. The main objective of this study was to establish a luminal epithelial cell-line from a lactating goat. Explant culture technique was used to produce primary cells from the mammary tissue of a 4-year-old lactating Saanen goat. The outgrowing cells were purified by selective trypsinization to remove fibroblast cells in 3-4 serial passages. The purified cell cultures exhibited cobblestone morphology, typical of the mammary epithelial cells, formed clear islands when plated in low density, and exhibited dome-shaped structures, if cultured for extended time. The cells stained positive with anti-human cytokeratin 18 antibodies, confirming their epithelial nature. Cell cultures also stained positive with rabbit anti-bovine &beta;-lactoglobulin antibodies, indicating milk production in these cells. The cell-line has potential as an in-vitro cell model to understand signaling during milk synthesis, mammary gland development, and testing DNA constructs for therapeutic protein secretion in milk, prior to production of transgenic goats.

scholarly journals Effects of a tart cherry (Prunus cerasus L.) phenolic extract on Porphyromonas gingivalis and its ability to impair the oral epithelial barrier

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0246194
Author(s):  
Amel Ben Lagha ◽  
Geneviève Pellerin ◽  
Katy Vaillancourt ◽  
Daniel Grenier
Keyword(s):  
Protective Effect ◽  
Porphyromonas Gingivalis ◽  
Periodontal Diseases ◽  
Cysteine Proteases ◽  
Prunus Cerasus ◽  
Epithelial Barrier ◽  
Tart Cherry ◽  
Phenolic Extract ◽  
Oral Epithelial

Periodontal diseases, including gingivitis and periodontitis, are a global oral health problem. Porphyromonas gingivalis, a key pathogen involved in the onset of periodontitis, is able to colonize the subgingival epithelium and invade the underlying connective tissue due to the contribution of cysteine proteases known as gingipains. In this study, we investigated the effects of a phenolic extract prepared from tart cherry (Prunus cerasus L.) juice on the growth, adherence, and protease activity of P. gingivalis. We also assessed the protective effect of the tart cherry extract on the disruption of the oral epithelial barrier induced by P. gingivalis. The tart cherry extract that contains procyanidins and quercetin and its derivatives (rutinoside, glucoside) as the most important phenolic compounds attenuated P. gingivalis growth, reduced adherence to an experimental basement membrane matrix model, and decreased the protease activities of P. gingivalis. The tart cherry extract also exerted a protective effect on the integrity of the oral epithelial barrier in an in vitro model infected with P. gingivalis. More specifically, the extract prevented a decrease in transepithelial electrical resistance as well as the destruction of tight junction proteins (zonula occludens-1 and occludin). These results suggest that the tart cherry phenolic extract may be a promising natural product for the treatment of periodontitis through its ability to attenuate the virulence properties of P. gingivalis and curtail the ability of this pathogen to impair the oral epithelial barrier.

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