Cranberry and Grape Juices Affect Tight Junction Function and Structural Integrity of Rotavirus-Infected Monkey Kidney Epithelial Cell Monolayers

Alveolar epithelial cells in patients with acute lung injury subjected to mechanical ventilation are exposed to increased procoagulant activity and mechanical strain. Thrombin induces epithelial cell stiffening, contraction, and cytoskeletal remodeling, potentially compromising the balance of forces at the alveolar epithelium during cell stretching. This balance can be further compromised by the loss of integrity of cell-cell junctions in the injured epithelium. The aim of this work was to study the effect of stretch on the structural integrity and micromechanics of human alveolar epithelial cell monolayers exposed to thrombin. Confluent and subconfluent cells (A549) were cultured on collagen-coated elastic substrates. After exposure to thrombin (0.5 U/ml), a stepwise cell stretch (20%) was applied with a vacuum-driven system mounted on an inverted microscope. The structural integrity of the cell monolayers was assessed by comparing intercellular and intracellular strains within the monolayer. Strain was measured by tracking beads tightly bound to the cell surface. Simultaneously, cell viscoelasticity was measured using optical magnetic twisting cytometry. In confluent cells, thrombin did not induce significant changes in transmission of strain from the substrate to overlying cells. By contrast, thrombin dramatically impaired the ability of subconfluent cells to follow imposed substrate deformation. Upon substrate unstretching, thrombin-treated subconfluent cells exhibited compressive strain (9%). Stretch increased stiffness (56–62%) and decreased cell hysteresivity (13–22%) of vehicle cells. By contrast, stretch did not increase stiffness of thrombin-treated cells, suggesting disruption of cytoskeletal structures. Our findings suggest that thrombin could exacerbate epithelial barrier dysfunction in injured lungs subjected to mechanical ventilation.

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Establishment of Intestinal Epithelial Cell Monolayers and Their Use in Calcium Switch Assay for Assessment of Intestinal Tight Junction Assembly

10.1007/7651_2021_347 ◽

2021 ◽

Author(s):

Pawin Pongkorpsakol ◽

Wilasinee Satianrapapong ◽

Preedajit Wongkrasant ◽

Peter R. Steinhagen ◽

Nuttha Tuangkijkul ◽

...

Keyword(s):

Epithelial Cell ◽

Tight Junction ◽

Intestinal Epithelial Cell ◽

Intestinal Epithelial ◽

Cell Monolayers ◽

Epithelial Cell Monolayers

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A protocol for imaging microvilli biogenesis on the surface of cultured porcine kidney epithelial cell monolayers

STAR Protocols ◽

10.1016/j.xpro.2021.100998 ◽

2021 ◽

Vol 2 (4) ◽

pp. 100998

Author(s):

Isabella M. Gaeta ◽

Leslie M. Meenderink ◽

Matthew J. Tyska

Keyword(s):

Epithelial Cell ◽

Porcine Kidney ◽

Cell Monolayers ◽

Kidney Epithelial Cell ◽

Epithelial Cell Monolayers

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Inflammatory cytokines induce apoptotic and necrotic cell shedding from human proximal tubular epithelial cell monolayers.

Kidney International ◽

10.1046/j.1523-1755.1999.00456.x ◽

1999 ◽

Vol 55 (6) ◽

pp. 2573 ◽

Cited By ~ 23

Author(s):

P.A. Glynne

Keyword(s):

Epithelial Cell ◽

Inflammatory Cytokines ◽

Tubular Epithelial Cell ◽

Necrotic Cell ◽

Cell Monolayers ◽

Proximal Tubular Epithelial Cell ◽

Proximal Tubular ◽

Epithelial Cell Monolayers ◽

Cell Shedding

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Induction of T-lymphocyte differentiation by thymic epithelial cell monolayers

Clinical Immunology and Immunopathology ◽

10.1016/0090-1229(78)90187-3 ◽

1978 ◽

Vol 10 (3) ◽

pp. 315-324 ◽

Cited By ~ 22

Author(s):

Judith I. Willis-Carr ◽

Hans D. Ochs ◽

Ralph J. Wedgwood

Keyword(s):

Epithelial Cell ◽

T Lymphocyte ◽

Thymic Epithelial Cell ◽

Lymphocyte Differentiation ◽

Cell Monolayers ◽

Epithelial Cell Monolayers

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Characteristics of Passive Solute Transport across Primary Rat Alveolar Epithelial Cell Monolayers

Membranes ◽

10.3390/membranes11050331 ◽

2021 ◽

Vol 11 (5) ◽

pp. 331

Author(s):

Yong Ho Kim ◽

Kwang-Jin Kim ◽

David Z. D’Argenio ◽

Edward D. Crandall

Keyword(s):

Epithelial Cell ◽

Tight Junctions ◽

Pore Radius ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial Cells ◽

Type I ◽

Cell Monolayers ◽

Alveolar Epithelial ◽

Epithelial Cell Monolayers ◽

Hydrophilic Solutes

Primary rat alveolar epithelial cell monolayers (RAECM) were grown without (type I cell-like phenotype, RAECM-I) or with (type II cell-like phenotype, RAECM-II) keratinocyte growth factor to assess passive transport of 11 hydrophilic solutes. We estimated apparent permeability (Papp) in the absence/presence of calcium chelator EGTA to determine the effects of perturbing tight junctions on “equivalent” pores. Papp across RAECM-I and -II in the absence of EGTA are similar and decrease as solute size increases. We modeled Papp of the hydrophilic solutes across RAECM-I/-II as taking place via heterogeneous populations of equivalent pores comprised of small (0.41/0.32 nm radius) and large (9.88/11.56 nm radius) pores, respectively. Total equivalent pore area is dominated by small equivalent pores (99.92–99.97%). The number of small and large equivalent pores in RAECM-I was 8.55 and 1.29 times greater, respectively, than those in RAECM-II. With EGTA, the large pore radius in RAECM-I/-II increased by 1.58/4.34 times and the small equivalent pore radius increased by 1.84/1.90 times, respectively. These results indicate that passive diffusion of hydrophilic solutes across an alveolar epithelium occurs via small and large equivalent pores, reflecting interactions of transmembrane proteins expressed in intercellular tight junctions of alveolar epithelial cells.

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A Novel Pharmacological Approach to Enhance the Integrity and Accelerate Restitution of the Intestinal Epithelial Barrier

Inflammatory Bowel Diseases ◽

10.1093/ibd/izaa063 ◽

2020 ◽

Vol 26 (9) ◽

pp. 1340-1352

Author(s):

Xuelei Cao ◽

Lei Sun ◽

Susana Lechuga ◽

Nayden G Naydenov ◽

Alex Feygin ◽

...

Keyword(s):

Wound Healing ◽

Epithelial Cell ◽

Epithelial Barrier ◽

Intestinal Epithelial ◽

Intestinal Epithelial Barrier ◽

Cell Monolayers ◽

Barrier Disruption ◽

Epithelial Cell Monolayers ◽

Immunofluorescence Labeling ◽

E Cadherin

Abstract Background Disruption of the gut barrier is an essential mechanism of inflammatory bowel diseases (IBDs) contributing to the development of mucosal inflammation. A hallmark of barrier disruption is the disassembly of epithelial adherens junctions (AJs) driven by decreased expression of a major AJ protein, E-cadherin. A group of isoxazole compounds, such as E-cadherin-upregulator (ECU) and ML327, were previously shown to stimulate E-cadherin expression in poorly differentiated human cancer cells. This study was designed to examine whether these isoxazole compounds can enhance and protect model intestinal epithelial barriers in vitro. Methods The study was conducted using T84, SK-CO15, and HT-29 human colonic epithelial cell monolayers. Disruption of the epithelial barrier was induced by pro-inflammatory cytokines, tumor necrosis factor-α, and interferon-γ. Barrier integrity and epithelial junction assembly was examined using different permeability assays, immunofluorescence labeling, and confocal microscopy. Epithelial restitution was analyzed using a scratch wound healing assay. Results E-cadherin-upregulator and ML327 treatment of intestinal epithelial cell monolayers resulted in several barrier-protective effects, including reduced steady-state epithelial permeability, inhibition of cytokine-induced barrier disruption and junction disassembly, and acceleration of epithelial wound healing. Surprisingly, these effects were not due to upregulation of E-cadherin expression but were mediated by multiple mechanisms including inhibition of junction protein endocytosis, attenuation of cytokine-induced apoptosis, and activation of promigratory Src and AKT signaling. Conclusions Our data highlight ECU and ML327 as promising compounds for developing new therapeutic strategies to protect the integrity and accelerate the restitution of the intestinal epithelial barrier in IBD and other inflammatory disorders.

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