Regulation of endothelial barrier function during flow-induced conversion to an arterial phenotype

Seebach, Jochen; Donnert, Gerald; Kronstein, Romy; Werth, Sebastian; Wójciak-Stothard, Beata; Falzarano, Darryl; Mrowietz, C.; Hell, Stefan W.; Schnittler, Hans

doi:10.1016/j.cardiores.2007.04.017

Cited by 80 publications

(87 citation statements)

References 62 publications

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“…26 In addition, the increase in t also caused translocation of cortactin to intercellular junctions within 15 min, and this was prevented by transfection of the cells with N17Rac1, a dominant negative form of Rac1. 34 In a different study, transfection of HUVEC with N17Rac1 prevented the initial increase in TER caused by a step increase in t. 10 In the current study, the actin stabilizer phalloidin inhibited the shear stress-induced barrier enhancement, demonstrating the importance of normal actin dynamics (Fig. 5).…”

Section: Discussionsupporting

confidence: 55%

“…Lymphatic endothelial cells responded to step increases in t by rapidly enhancing their barrier function in a similar manner as reported for endothelial cells from porcine pulmonary trunks (PSEC), 9 human umbilical vein endothelial cells (HUVEC), 10 and bovine aortic endothelial cells (BAEC). 8,30 In addition, the magnitude of the elevation of HMLEC-d TER was directly related to the degree of the increase in t, similar to a previous finding with PSEC.…”

Section: Discussionsupporting

confidence: 55%

“…11 In cultured vascular endothelial cell models, increasing t causes an initial rapid increase in barrier function, followed by decrease in barrier function over longer periods of time. [8][9][10] In both of the isolated venule and cultured endothelial cell models, the effects of increased t were reversed upon returning to the baseline t. [8][9][10][11] These studies demonstrate that shear stress sensing is an important property of vascular endothelium for regulating barrier function.…”

Section: Introductionmentioning

confidence: 81%

“…7 In blood vessels and in cultured vascular endothelial cell models, changes in the laminar fluid flow rate can influence barrier function. [8][9][10][11] For example, increasing the fluid flow through isolated, perfused coronary venules not only increases filtration of solutes, but also increases their diffusive permeability coefficient to albumin through a NO-related mechanism. 11 In cultured vascular endothelial cell models, increasing t causes an initial rapid increase in barrier function, followed by decrease in barrier function over longer periods of time.…”

Section: Introductionmentioning

confidence: 99%

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Lymphatic Endothelial Cells Adapt Their Barrier Function in Response to Changes in Shear Stress

Breslin

Kurtz

2009

Lymphatic Research and Biology

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Background: Lymphatic endothelial cells form an important barrier necessary for normal lymph formation and propulsion. However, little is known about how physical forces within lymphatic vessels affect endothelial barrier function. The purpose of this study was to characterize how laminar flow affects lymphatic endothelial barrier function and to test whether endothelial cells respond to flow changes by activating the intracellular actin cytoskeleton to enhance barrier function. Methods and Results: Cultured adult human dermal microlymphatic endothelial cells (HMLEC-d) were grown on small gold electrodes arranged within a flow channel, and transendothelial electrical resistance (TER), an index of barrier function, was determined. Laminar flow was applied to the cells at a baseline shear stress of 0.5 dynes=cm 2 , and was increased to 2.5, 5.0, or 9.0 dynes=cm 2 , causing a magnitude-dependent increase in barrier function that was reversed 30 min later when the shear stress was returned to baseline. This response was abolished by blockade of actin dynamics with 10 mM phalloidin, and significantly inhibited by blockade of Rac1 activity with 50 mM NSC23766. Blockade of protein kinase A (10 mM H-89) did not inhibit the response. Mathematical modeling based on our impedance data showed that the flow-induced changes in TER were primarily due to altered current flow between cells and not beneath cells. Conclusions: These results suggest that lymphatic endothelial cells dynamically alter their morphology and barrier function in response to changes in shear stress by a mechanism dependent upon Rac1-mediated actin dynamics.

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Section: Discussionsupporting

confidence: 55%

Section: Discussionsupporting

confidence: 55%

Section: Introductionmentioning

confidence: 81%

Section: Introductionmentioning

confidence: 99%

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Lymphatic Endothelial Cells Adapt Their Barrier Function in Response to Changes in Shear Stress

Breslin

Kurtz

2009

Lymphatic Research and Biology

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“…Similar permeability characteristics have been reported previously for endothelial cells when they are exposed to FSS, where the permeability increases immediately at the onset of flow and slowly plateaus to a baseline value. 42,43 The introduction of flow does bring about a transient increase in permeability for the BAOEC layer that stabilizes after about 2.5 h of flow. Thus, it is safe to assume that this study is performed on a BAOEC layer that has been primed to flow, and the study performed after 6 h of exposure to FSS will not be affected by any initial flow related changes in permeability.…”

Section: Resultsmentioning

confidence: 99%

Characterization of vascular permeability using a biomimetic microfluidic blood vessel model

et al. 2017

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The inflammatory response in endothelial cells (ECs) leads to an increase in vascular permeability through the formation of gaps. However, the dynamic nature of vascular permeability and external factors involved is still elusive. In this work, we use a biomimetic blood vessel (BBV) microfluidic model to measure in real-time the change in permeability of the EC layer under culture in physiologically relevant flow conditions. This platform studies the dynamics and characterizes vascular permeability when the EC layer is triggered with an inflammatory agent using tracer molecules of three different sizes, and the results are compared to a transwell insert study. We also apply an analytical model to compare the permeability data from the different tracer molecules to understand the physiological and bio-transport significance of endothelial permeability based on the molecule of interest. A computational model of the BBV model is also built to understand the factors influencing transport of molecules of different sizes under flow. The endothelial monolayer cultured under flow in the BBV model was treated with thrombin, a serine protease that induces a rapid and reversible increase in endothelium permeability. On analysis of permeability data, it is found that the transport characteristics for fluorescein isothiocyanate (FITC) dye and FITC Dextran 4k Da molecules are similar in both BBV and transwell models, but FITC Dextran 70k Da molecules show increased permeability in the BBV model as convection flow (Peclet number > 1) influences the molecule transport in the BBV model. We also calculated from permeability data the relative increase in intercellular gap area during thrombin treatment for ECs in the BBV and transwell insert models to be between 12% and 15%. This relative increase was found to be within range of what we quantified from F-actin stained EC layer images. The work highlights the importance of incorporating flow in in vitro vascular models, especially in studies involving transport of large size objects such as antibodies, proteins, nano/micro particles, and cells. Published by AIP Publishing.

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Microfluidic blood–brain barrier model provides in vivo‐like barrier properties for drug permeability screening

Wang

Abaci

Shuler

2016

Biotech & Bioengineering

410

389

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Efficient delivery of therapeutics across the neuroprotective blood-brain barrier (BBB) remains a formidable challenge for central nervous system drug development. Highfidelity in vitro models of the BBB could facilitate effective early screening of drug candidates targeting the brain. In this study, we developed a microfluidic BBB model that is capable of mimicking in vivo BBB characteristics for a prolonged period and allows for reliable in vitro drug permeability studies under recirculating perfusion. We derived brain microvascular endothelial cells (BMECs) from human induced pluripotent stem cells (hiPSCs) and cocultured them with rat primary astrocytes on the two sides of a porous membrane on a pumpless microfluidic platform for up to 10 days. The microfluidic system was designed based on the blood residence time in human brain tissues, allowing for medium recirculation at physiologically relevant perfusion rates with no pumps or external tubing, meanwhile minimizing wall shear stress to test whether shear stress is required for in vivo-like barrier properties in a microfluidic BBB model. This BBB-on-a-chip model achieved significant barrier integrity as evident by continuous tight junction formation and in vivo-like values of trans-endothelial electrical resistance (TEER). The TEER levels peaked above 4000 V Á cm 2 on day 3 on chip and were sustained above 2000 V Á cm 2 up to 10 days, which are the highest sustained TEER values reported in a microfluidic model. We evaluated the capacity of our microfluidic BBB model to be used for drug permeability studies using large molecules (FITC-dextrans) and model drugs (caffeine, cimetidine, and doxorubicin). Our analyses demonstrated that the permeability coefficients measured using our model were comparable to in vivo values. Our BBB-on-a-chip model closely mimics physiological BBB barrier functions and will be a valuable tool for screening of drug candidates. The residence time-based design of a microfluidic platform will enable integration with other organ modules to simulate multi-organ interactions on drug response.

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Regulation of endothelial barrier function during flow-induced conversion to an arterial phenotype

Cited by 80 publications

References 62 publications

Lymphatic Endothelial Cells Adapt Their Barrier Function in Response to Changes in Shear Stress

Lymphatic Endothelial Cells Adapt Their Barrier Function in Response to Changes in Shear Stress

Characterization of vascular permeability using a biomimetic microfluidic blood vessel model

Microfluidic blood–brain barrier model provides in vivo‐like barrier properties for drug permeability screening

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