Permeability characteristics of human endothelial monolayers seeded on different extracellular matrix proteins

Abstract

OBJECTIVE: To investigate whether endothelial monolayer permeability changes induced by inflammatory mediators are affected by the extracellular matrix protein used for cell seeding. METHODS: Human umbilical venular endothelial cells (HUVEC) were grown to confluent monolayers on membranes coated with either collagen, fibronectin or gelatin. The permeability to albumin and dextran was then assessed, both under normal conditions and after treatment with tumor necrosis factor-alpha (TNF-alpha) and bacterial lipopolysaccharide (LPS). RESULTS: With any of the three protein coatings, tight junctions were formed all over the monolayers. The permeability of the coated membranes to albumin and dextran was reduced strongly by confluent monolayers; the relative reduction was similar for the three matrix proteins used. Pre-incubation of the monolayers with either TNF-alpha or LPS increased permeability dose dependently. However, the relative increase due to either treatment was independent of the protein used for membrane coating. CONCLUSION: The extracellular matrix protein used for initial seeding of endothelial cultures plays a minor role in determining the permeability changes induced in HUVEC monolayers by inflammatory mediators.

Acute reactive oxygen species (ROS)-dependent effects of IL-1β, TNF-α, and IL-6 on the glomerular filtration barrier (GFB) in vivo

“…The mechanisms through which cytokines exert their acute effects on the endothelium to increase its permeability are not fully understood. Studies on models of endothelial monolayers in vitro have suggested that multiple mechanisms are involved, including Ca 2ϩ signaling, endothelial cell contraction with retraction of cells, and changes in intercellular junctions, producing interendothelial gaps (40). Cytokines are also thought to induce production of several other inflammatory mediators and to activate enzymes.…”

“…Ex vivo studies have shown that endothelial permeability can increase by exposing endothelial cells to TNF-␣ or IL-1␤ (14,34,40,43). IL-6 has also been shown to increase endothelial permeability through alterations of tight junctions, and by rearrangement of intracellular F-actin and changes in cell shape (21,35), which, however, could not be confirmed in other studies (14).…”

Barrier Protective Activities of Phloroglucinol on Lipopolysaccharide (LPS)-Induced Barrier Disruption in Human Endothelial Cells

“…Monolayers of human endothelial cells forming the innermost layer of vessels presents a biological barrier to permeation of liquid, proteins, and solutes from the intravascular compartment to the interstitium [2,3]. The vascular endothelium is composed of a monolayer of vascular endothelial cells.…”

“…Evidence suggests that the interaction between cell adhesion molecules (CAMs) and endothelial cells can contribute to pro-inflammatory responses by recruiting monocytes to the injury sites [3]. Inflammatory mediators, such as thrombin, bradykinin, histamine, lipopolysaccharide (LPS), and others, upon binding to their receptors, disrupt the organization of inter-endothelial junctions, thereby opening the junctional barrier [4].…”

Dengue Virus neither Directly Mediates Hyperpermeability nor Enhances Tumor Necrosis Factor-^|^alpha;-Induced Permeability In Vitro

“…In addition, ECs from different tissue origins, such as LSEC-1, HMEC-1, or HPMEC-ST1.6R, which may have different characteristics, responded differently to DENV infection (9,15,31,32). In the present study, primary HUVECs isolated from normal human umbilical vein were evaluated because they are more relevant to the pathophysiology of DHF/DSS and the most commonly used model to investigate macromolecule transport (33,34). Here we showed that all DENV-2 strains were able to infect ECs and replicate actively (Fig.…”