Collagen-mimetic hydrogels promote human endothelial cell adhesion, migration and phenotypic maturation

Monocyte-endothelial cell adhesion is a key early event in atherogenesis. C-reactive protein (CRP), a cardiovascular risk marker, is known to stimulate ICAM and VCAM in human aortic endothelial cells (HAEC) and induces monocyte-endothelial cell adhesion. In this study, we examined the mechanisms by which native CRP promotes monocyte-endothelial cell adhesion under static conditions and tested the effect of CRP on adhesion under shear flow. Incubation of HAEC with CRP (>25 μg/ml) upregulated NF-κB activity, and this resulted in a significant increase in ICAM (54% increase, P < 0.001), VCAM (41% increase, P < 0.01), and monocyte-endothelial cell adhesion (44% increase, P < 0.02) compared with those of control. Preincubation with antibodies to CD32 and CD64 but not CD16 effectively inhibited this activation. Blocking NF-κB activity with inhibitors or a dominant negative inhibitory κB significantly decreased ICAM, VCAM upregulation, and subsequent monocyte-endothelial cell adhesion. Preincubation with antibodies to CD32 and CD64 or transient transfection with small interference RNA to CD32 attenuated CRP-induced NF-κB activity, ICAM, VCAM, and monocyte-endothelial cell adhesion under static conditions. Also, the Syk kinase inhibitor piceatannol and MG-132, a proteasome degradation inhibitor, produced similar attenuation in NF-κB activity, ICAM, VCAM, and adhesion. Furthermore, CRP-activated endothelial cells supported monocyte rolling, arrest, and transmigration in shear flow (2 dyn/cm2), and this was also inhibited by preincubation with antibodies to CD32 and CD64. Thus, in HAEC, CRP upregulates monocyte-endothelial adhesion by activation of NF-κB through engaging the Fcγ receptors CD32 and CD64.

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Asiaticoside reduces TNF-α-induced increased soluble platelet endothelial cell adhesion molecule -1 (sPECAM-1) levels in human aortic endothelial cells

Planta Medica ◽

10.1055/s-0035-1565739 ◽

2015 ◽

Vol 81 (16) ◽

Author(s):

LY Fong ◽

CT Ng ◽

Z Ahmad

Keyword(s):

Endothelial Cells ◽

Cell Adhesion ◽

Endothelial Cell ◽

Cell Adhesion Molecule ◽

Aortic Endothelial Cells ◽

Platelet Endothelial Cell Adhesion ◽

Tnf Α ◽

Cell Adhesion Molecule 1 ◽

Human Aortic Endothelial Cells ◽

Endothelial Cell Adhesion

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Differential effects of laminin, intact type IV collagen, and specific domains of type IV collagen on endothelial cell adhesion and migration.

The Journal of Cell Biology ◽

10.1083/jcb.106.4.1365 ◽

1988 ◽

Vol 106 (4) ◽

pp. 1365-1373 ◽

Cited By ~ 143

Author(s):

T J Herbst ◽

J B McCarthy ◽

E C Tsilibary ◽

L T Furcht

Keyword(s):

Endothelial Cells ◽

Cell Adhesion ◽

Endothelial Cell ◽

Type Iv Collagen ◽

Aortic Endothelial Cells ◽

Aortic Endothelial Cell ◽

Type Iv ◽

And Migration ◽

Endothelial Cell Adhesion ◽

Aortic Endothelial

Laminin and type IV collagen were compared for the ability to promote aortic endothelial cell adhesion and directed migration in vitro. Substratum-adsorbed IV promoted aortic endothelial cell adhesion in a concentration dependent fashion attaining a maximum level 141-fold greater than controls within 30 min. Aortic endothelial cell adhesion to type IV collagen was not inhibited by high levels (10(-3) M) of arginyl-glycyl-aspartyl-serine. In contrast, adhesion of aortic endothelial cells on laminin was slower, attaining only 53% of the adhesion observed on type IV collagen by 90 min. Type IV collagen when added to the lower well of a Boyden chamber stimulated the directional migration of aortic endothelial cells in a concentration dependent manner with a maximal response 6.9-fold over control levels, whereas aortic endothelial cells did not migrate in response to laminin at any concentration (.01-2.0 X 10(-7) M). Triple helix-rich fragments of type IV collagen were nearly as active as intact type IV collagen in stimulating both adhesion and migration whereas the carboxy terminal globular domain was less active at promoting adhesion (36% of the adhesion promoted by intact type IV collagen) or migration. Importantly, aortic endothelial cells also migrate to substratum adsorbed gradients of type IV collagen suggesting that the mechanism of migration is haptotactic in nature. These results demonstrate that the aortic endothelial cell adhesion and migration is preferentially promoted by type IV collagen compared with laminin, and has a complex molecular basis which may be important in angiogenesis and large vessel repair.

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7-ketocholesterol treatment modulates expression of the cell adhesion molecules in human aortic endothelial cells-pilot study

Atherosclerosis ◽

10.1016/j.atherosclerosis.2017.06.430 ◽

2017 ◽

Vol 263 ◽

pp. e135

Author(s):

Matej Vicen ◽

Michala Varejckova ◽

Radim Havelek ◽

Barbora Vitverová ◽

Petra Fikrová ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Adhesion ◽

Pilot Study ◽

Adhesion Molecules ◽

Cell Adhesion Molecules ◽

Aortic Endothelial Cells ◽

Human Aortic Endothelial Cells ◽

Aortic Endothelial

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Polyploidy impairs human aortic endothelial cell function and is prevented by nicotinamide phosphoribosyltransferase

AJP Cell Physiology ◽

10.1152/ajpcell.00357.2009 ◽

2010 ◽

Vol 298 (1) ◽

pp. C66-C74 ◽

Cited By ~ 25

Author(s):

Nica M. Borradaile ◽

J. Geoffrey Pickering

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Replicative Senescence ◽

Cell Function ◽

Human Aortic Endothelial Cell ◽

Nicotinamide Phosphoribosyltransferase ◽

Endothelial Cell Function ◽

Aortic Endothelial Cells ◽

Human Aortic Endothelial Cells ◽

Aortic Endothelial

Polyploid endothelial cells are found in aged and atherosclerotic arteries. However, whether increased chromosome content has an impact on endothelial cell function is unknown. We show here that human aortic endothelial cells become tetraploid as they approach replicative senescence. Furthermore, accumulation of tetraploid endothelial cells was accelerated during growth in high glucose. Interestingly, induction of polyploidy was completely prevented by modest overexpression of the NAD+ regenerating enzyme, nicotinamide phosphoribosyltransferase (Nampt). To determine the impact of polyploidy on endothelial cell function, independent of replicative senescence, we induced tetraploidy using the spindle poison, nocodazole. Global gene expression analyses of tetraploid endothelial cells revealed a dysfunctional phenotype characterized by a cell cycle arrest profile (decreased CCNE2/A2, RBL1, BUB1B; increased CDKN1A) and increased expression of genes involved in inflammation ( IL32, TNFRSF21/10C, PTGS1) and extracellular matrix remodeling ( COL5A1, FN1, MMP10/14). The protection from polyploidy conferred by Nampt was not associated with enhanced poly(ADP-ribose) polymerase-1 or sirtuin (SIRT) 2 activity, but with increased SIRT1 activity, which reduced cellular reactive oxygen species and the associated oxidative stress stimulus for the induction of polyploidy. We conclude that human aortic endothelial cells are prone to chromosome duplication that, in and of itself, can induce characteristics of endothelial dysfunction. Moreover, the emergence of polyploid endothelial cells during replicative aging and glucose overload can be prevented by optimizing the Nampt-SIRT1 axis.

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Silicon microgrooves for contact guidance of human aortic endothelial cells

Beilstein Journal of Nanotechnology ◽

10.3762/bjnano.8.72 ◽

2017 ◽

Vol 8 ◽

pp. 675-681 ◽

Cited By ~ 4

Author(s):

Sara Fernández-Castillejo ◽

Pilar Formentín ◽

Úrsula Catalán ◽

Josep Pallarès ◽

Lluís F Marsal ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Adhesion ◽

Cellular Response ◽

Cytotoxic Effect ◽

Contact Guidance ◽

Silicon Substrates ◽

Aortic Endothelial Cells ◽

Human Aortic Endothelial Cells ◽

Microscopy Images ◽

Aortic Endothelial

Background: Micro- and nanoscale substrates have been fabricated in order to study the influence of the topography on the cellular response. The aim of this work was to prepare different collagen-coated silicon substrates displaying grooves and ridges to mimic the aligned and elongated endothelium found in linear vessels, and to use them as substrates to study cell growth and behaviour. Results: The influence of groove-shaped substrates on cell adhesion, morphology and proliferation were assessed, by comparing them to flat silicon substrates, used as control condition. Using human aortic endothelial cells, microscopy images demonstrate that the cellular response is different depending on the silicon surface, when it comes to cell adhesion, morphology (alignment, circularity and filopodia presence) and proliferation. Moreover, these structures exerted no cytotoxic effect. Conclusion: The results suggest that topographical patterning influences cell response. Silicon groove substrates can be used in developing medical devices with microscale features to mimic the endothelium in lineal vessels.

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Differential activation of NF-kappa B in human aortic endothelial cells conditioned to specific flow environments

AJP Cell Physiology ◽

10.1152/ajpcell.1997.273.2.c572 ◽

1997 ◽

Vol 273 (2) ◽

pp. C572-C578 ◽

Cited By ~ 111

Author(s):

S. Mohan ◽

N. Mohan ◽

E. A. Sprague

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

High Shear ◽

Monocyte Adhesion ◽

Nf Kappa B ◽

Differential Activation ◽

Aortic Endothelial Cells ◽

Human Aortic Endothelial Cells ◽

Low Shear ◽

Aortic Endothelial

Endothelial cell-monocyte interaction plays an important role in atherogenesis. The expressions of some endothelial cell adhesion molecules involved in endothelial cell-monocyte interactions are regulated by transcription factor NF-kappa B. Because low shear stress has been known to influence endothelial monocyte adhesion, the differential activation of NF-kappa B under different flow regimens across time (0.5-24 h) was investigated. Nuclear proteins from flow-conditioned human aortic endothelial cells (HAEC) were analyzed by electrophoretic mobility shift assay using [gamma-32P]dATP-labeled NF-kappa B-specific oligonucleotide. Our results demonstrated that NF-kappa B activation was significantly elevated in HAEC exposed to prolonged (> 2 h) steady low shear (2 dyn/cm2) and pulsatile low shear (2 +/- 2 dyn/cm2) compared with HAEC exposed to high shear (16 dyn/cm2). In contrast, at 30 min, high shear-exposed HAEC exhibited an early, transient increase in NF-kappa B activity, relative to low shear-exposed cells, which reversed on continued exposure to high shear. Maximum activity in both low shear- and pulsatile low shear-conditioned HAEC was observed at 16 h compared with HAEC exposed to prolonged high shear. These results indicate that exposure of HAEC to prolonged low shear conditions is associated with significantly increased and prolonged NF-kappa B activity. This observation might provide a mechanism to explain the increased monocyte adhesion in atherosclerosisprone arterial sites exposed to chronic low-shear flow patterns.

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