Regions Of Low Shear Stress Research Articles

Effect of shear stress on vascular inflammation and plaque development

This review describes evidence that shear stress acts through modulation of inflammation and by that process affects atherogenesis and plaque composition. In low shear stress regions antiatherogenic transcription factors are downregulated and pro-atherogenic transcription factors are upregulated. Consequently, inflammatory cells may home low shear stress regions more easily to the plaques because of increased expression of adhesion factors, a decreased rolling speed and an increased expression of chemokines, thereby changing the composition of the plaques into a more vulnerable phenotype. In contrast, in advanced plaque development vascular lumen decreases and shear stress increases, especially upstream of the plaques. The predominant upstream location of lipids induces a prevalent upstream location of inflammatory cells leading to localized plaque rupture. Shear stress has been shown to play a role in plaque induction, plaque progression and plaque rupture. The mechanism for plaque induction seems to differ from the role of shear stress for plaque rupture, whereby the former mechanism is induced by low shear stress and the latter by high shear stress.

Flux enhancement and cake formation in air-sparged cross-flow microfiltration

Abstract The flux enhancement in cross-flow microfiltration of submicron particles by sparged air-bubble is studied. The effects of operating conditions, such as air-bubble velocity, suspension velocity and filtration pressure, on the cake properties and filtration flux are discussed thoroughly. The results show that the pseudo-steady filtration flux increases as the air-bubble velocity and filtration pressure increase. The sparged air-bubble can significantly improve filtration flux, but the flux enhancement is more remarkable in the lower air-bubble velocity region. A gas–liquid two-phase flow model is adopted for estimating the shear stress acting on the membrane surface under various operating conditions. The cake mass can be significantly reduced by increasing the shear stress acting on the membrane surface. However, the SEM analysis illustrates that the particle packing structure becomes more compact as the air-bubble velocity increases. This results in a slightly higher average specific cake filtration resistance under higher air-bubble velocity. Consequently, a minimum flux occurs at the critical shear stress, e.g., τ w = 1.1 N/m 2 in this study, when these effects are both taken into consideration. As the shear stress increases by increasing the suspension or gas-bubble velocity, the filtration flux decreases in the low shear stress region but, on the contrary, quickly increases in the high shear stress region. Furthermore, a force balance model is derived for understanding the particle deposition on the membrane surface. The relationship among filtration flux, shear stress and overall filtration resistance is obtained and verified by experimental data.

Plaque Morphology in High Versus Low Shear Stress Regions of Human Carotid Stenosis

Plaque Morphology in High Versus Low Shear Stress Regions of Human Carotid Stenosis

Regions of low endothelial shear stress are the sites where coronary plaque progresses and vascular remodelling occurs in humans: an in vivo serial study

We performed serial intracoronary studies of patients with stable coronary artery disease (CAD) to investigate the relationships among baseline endothelial shear stress (ESS), CAD progression, and vascular remodelling. Local haemodynamic factors are critical determinants of plaque progression, vascular remodelling, and clinical CAD manifestations. The 3-D anatomy of coronary arteries with lumen obstruction <50% was determined by fusing intracoronary ultrasound and angiographic images in 13 patients at baseline and 8 +/- 2 months later. Cross-sectional area of plaque, lumen, and external elastic membrane (EEM), and coronary flow were measured. Local ESS was calculated. Subsegments with similar ESS were categorized based on low (<12 dynes/cm(2)) and moderate/higher ESS (> or =12 dynes/cm(2)). There were 47 subsegments of similar baseline ESS: nine with low ESS and 38 with moderate/higher ESS. Median subsegment length was 6.9 mm (25th-75th percentiles = 4.2-12.0), and median area of similar ESS of 52.6 mm(2) (25th-75th percentiles = 26.9-88.0). Subsegments with low ESS exhibited plaque progression when compared with subsegments with moderate/higher ESS (33.3% vs. 7.9%, respectively, P = 0.009 adjusted for clustering of lesions within patients) and constrictive remodelling (44.0% vs. 5.3%, respectively, P = 0.16 adjusted for clustering of lesions within patients). Expansive remodelling occurred with similar frequency in subsegments with low vs. moderate/higher baseline ESS. Plaque progresses in subsegments with low ESS, associated with either constrictive or expansive remodelling. Different mechanisms are likely responsible for expansive remodelling in different local vascular environments. Early in vivo identification of arterial subsegments likely to develop high-risk plaque characteristics may allow for selective interventions to avoid adverse cardiac outcomes.

Shear stress-induced changes in atherosclerotic plaque composition are modulated by chemokines.

We previously found that low shear stress (LSS) induces atherosclerotic plaques in mice with increased lipid and matrix metalloproteinase content and decreased vascular smooth muscle and collagen content. Here, we evaluated the role of chemokines in this process, using an extravascular device inducing regions of LSS, high shear stress, and oscillatory shear stress (OSS) in the carotid artery. One week of shear stress alterations induced expression of IFN-gamma-inducible protein-10 (IP-10) exclusively in the LSS region, whereas monocyte chemoattractant protein-1 (MCP-1) and the mouse homolog of growth-regulated oncogene alpha (GRO-alpha) were equally upregulated in both LSS and OSS regions. After 3 weeks, GRO-alpha and IP-10 were specifically upregulated in LSS regions. After 9 weeks, lesions with thinner fibrous caps and larger necrotic cores were found in the LSS region compared with the OSS region. Equal levels of MCP-1 expression were observed in both regions, while expression of fractalkine was found in the LSS region only. Blockage of fractalkine inhibited plaque growth and resulted in striking differences in plaque composition in the LSS region. We conclude that LSS or OSS triggers expression of chemokines involved in atherogenesis. Fractalkine upregulation is critically important for the composition of LSS-induced atherosclerotic lesions.

A new imaging technique to study 3-D plaque and shear stress distribution in human coronary artery bifurcations in vivo

Objective Bifurcations of coronary arteries are predilection sites for atherosclerosis and expansive remodeling, the latter being associated with plaque vulnerability. Both are related to blood flow-induced shear stress (SS). We present a new approach to generate 3-D reconstructions of coronary artery bifurcations in vivo and investigate the relationship between SS, wall thickness (WT) and remodeling. Methods The patient specific 3-D reconstruction of the main branch of the bifurcation was obtained by combining intravascular ultrasound and biplane angiography, and the 3-D lumen of the side branch was based on biplane angiography only. The two data sets were fused and computational methods were applied to determine the SS distribution, using patient derived flow and viscosity data. The intravascular ultrasound data allowed us to measure local WT and remodeling in the main branch. Results The lumen reconstruction procedure was successful and it was shown that the impact of the side branch on SS distribution in the main branch diminished within 3 mm. Distal to the bifurcation, two continuous regions in the main branch were identified. In the proximal region, we observed lumen preservation, and expansive remodeling. Although a plaque was observed in the low SS region at the non-divider wall, no relationship between SS and WT was found. In the distal region, we observed lumen narrowing and a significant positive relationship between SS and WT. Conclusions A new imaging technique was applied to generate a 3-D reconstruction of a human coronary artery bifurcation in vivo. The observed relationship between SS, WT and remodeling in this specific patient illustrates the spatial heterogeneity of the atherosclerosis in the vicinity of arterial bifurcations.

The role of endothelial calcium and nitric oxide in the localisation of atherosclerosis

A mathematical model of endothelial cell calcium signalling and nitric oxide synthesis under flow conditions is presented. The model is coupled to two important environmental stimuli for endothelial cells: the frictional shear stress exerted on the cell membrane by the blood flow; and the binding of adenosine triphosphate in the bloodstream to cell surface receptors. These stimuli are closely linked to haemodynamic flow conditions and are, in general, spatially varying, allowing the cellular response in different regions of the endothelium to be evaluated. This is used to indicate which areas of the artery wall experience reduced bioavailability of nitric oxide, which is a major factor in the onset of atherosclerosis. The model thus directly addresses the key issue of the causative link, and its underlying biochemical mechanisms, between incidence of atherosclerosis and regions of low wall shear stress (WSS). Model results show that intracellular levels of free calcium and endothelial nitric oxide synthase are lower in endothelial cells adjacent to a region of recirculating flow than in cells adjacent to regions of fully developed arterial flow. This will lead to deficient levels of nitric oxide in the recirculation zone and hence a potentially elevated risk of developing atherosclerotic plaque. This is consistent with the observed spatial correlation between atherosclerosis and regions of disturbed blood flow and low WSS, and provides a mechanism for the localisation of the disease to sites such as arterial bifurcations and bends.

Estimating the Hemodynamic Impact of Interventional Treatments of Aneurysms: Numerical Simulation with Experimental Validation: Technical Case Report

The goal of this study was to use phase-contrast magnetic resonance imaging and computational fluid dynamics to estimate the hemodynamic outcome that might result from different interventional options for treating a patient with a giant fusiform aneurysm. We followed a group of patients with giant intracranial aneurysms who have no clear surgical options. One patient demonstrated dramatic aneurysm growth and was selected for further analysis. The aneurysm geometry and input and output flow conditions were measured with contrast-enhanced magnetic resonance angiography and phase-contrast magnetic resonance imaging. The data was imported into a computational fluid dynamics program and the velocity fields and wall shear stress distributions were calculated for the presenting physiological condition and for cases in which the opposing vertebral arteries were either occluded or opened. These models were validated with in vitro flow experiments using a geometrically exact silicone flow phantom. Simulation indicated that altering the flow ratio in the two vertebrals would deflect the main blood jet into the aneurysm belly, and that this would likely reduce the extent of the region of low wall shear stress in the growth zone. Computational fluid dynamics flow simulations in a complex patient-specific aneurysm geometry were validated by in vivo and in vitro phase-contrast magnetic resonance imaging, and were shown to be useful in modeling the likely hemodynamic impact of interventional treatment of the aneurysm.

Micromolding of photocrosslinkable chitosan hydrogel for spheroid microarray and co-cultures

Bioengineering approaches, such as co-cultures of multiple cell types, that aim to mimic the physiological microenvironment may be beneficial for optimizing cell function and for engineering tissues in vitro. This study describes a novel method for preparing a spheroid microarray on microfabricated hydrogels, alone or in co-cultures. Photocrosslinkable chitosan was synthesized and utilized for fabricating hydrogel microstructures through a micromolding process. The chitosan surface was initially cell repellent but became increasingly cell adhesive over time. By using this unique property of chitosan hydrogels, it was possible to generate patterned co-cultures of spheroids and support cells. In this scheme, cells were initially microarrayed within low shear stress regions of microwells. Human hepatoblastoma cells, Hep G2, seeded in these wells formed spheroids with controlled sizes and shapes and stably secreted albumin during the culture period. The change of cell adhesive properties in the chitosan surface facilitated the adhesion and growth of a second cell type, NIH-3T3 fibroblast, and therefore enabled co-cultures of hepatocyte spheroids and fibroblast monolayers. This co-culture system could be a useful platform for studying heterotypic cell-cell interactions, for drug screening, and for developing implantable bioartificial organs.

Micromolding of photocrosslinkable hyaluronic acid for cell encapsulation and entrapment

Micropatterning of hydrogels is potentially useful for a variety of applications, including tissue engineering, fundamental biological studies, diagnostics, and high-throughput screening. Although synthetic polymers have been developed for these applications, natural polymers such as polysaccharides may have advantages for biological samples and cell-based devices because they are natural components of the in vivo microenvironment. In this study, we synthesized and used hyaluronic acid (HA) modified with photoreactive methacrylates to fabricate microstructures as functional components of microfabricated devices. To demonstrate the universality of this approach, two types of microstructures were formed. In the first approach, HA microstructures were fabricated and used as docking templates to enable the subsequent formation of cell microarrays within low shear stress regions of the patterns. Cells within these microwells remained viable, could generate spheroids, and could be retrieved using mechanical disruption. In the second approach, cells were encapsulated directly within the HA hydrogels. Arrays of viable embryonic stem (ES) cells or fibroblasts were encapsulated within HA hydrogels and could later be recovered using enzymatic digestion of the microstructures. These results demonstrate that it is possible to incorporate photocrosslinkable HA, a natural, versatile, degradable, and biocompatible biopolymer, into micro-electromechanical systems.

KLF2 provokes a gene expression pattern that establishes functional quiescent differentiation of the endothelium

AbstractThe flow-responsive transcription factor KLF2 is acquiring a leading role in the regulation of endothelial cell gene expression. A genome-wide microarray expression profiling is described employing lentivirus-mediated, 7-day overexpression of human KLF2 at levels observed under prolonged flow. KLF2 is not involved in lineage typing, as 42 endothelial-specific markers were unaffected. Rather, KLF2 generates a gene transcription profile (> 1000 genes) affecting key functional pathways such as cell migration, vasomotor function, inflammation, and hemostasis and induces a morphology change typical for shear exposure including stress fiber formation. Protein levels for thrombomodulin, endothelial nitric oxide synthase, and plasminogen activator inhibitor type-1 are altered to atheroprotective levels, even in the presence of the inflammatory cytokine TNF-α. KLF2 attenuates cell migration by affecting multiple genes including VEGFR2 and the potent antimigratory SEMA3F. The distribution of Weibel-Palade bodies in cultured cell populations is normalized at the single-cell level without interfering with their regulated, RalA-dependent release. In contrast, thrombin-induced release of Weibel-Palade bodies is significantly attenuated, consistent with the proposed role of VWF release at low–shear stress regions of the vasculature in atherosclerosis. These results establish that KLF2 acts as a central transcriptional switch point between the quiescent and activated states of the adult endothelial cell.

Effects of Arterial Bifurcation Geometry on Nucleotide Concentration at the Endothelium

A mathematical and computational analysis of nucleotide concentration for a two-dimensional artery bifurcation has been developed. This region of the vasculature is known to be exposed to spatially varying wall shear stress (WSS), hence the variation of adenosine nucleotides is of interest. A previously derived similarity solution for mass transport in blood boundary layers for arbitrary wall shear stress function has been used. For the analytical model, the geometric condition has been incorporated into the system using the theory for flow past a wedge. As the bifurcation angle varies different characteristics are exhibited, such as maxima in ADP concentration and the existence of a low wall shear stress region around the stagnation point, for large angles. In the limiting case of two-dimensional stagnation point flow the concentration was constant throughout the domain length. The computational simulations provided a more detailed understanding into how nucleotides vary. Similarities existed between the two solution methods in the vicinity of the stagnation point, but deviated as the fully developed condition prevailed. Additionally, the effect of pulsatile flow has been included, leading to considerable gradients in wall shear stress, both temporal and spatial. However, the resulting nucleotide concentration is determined by the time-averaged wall shear stress. The effects of flow-induced ATP release have also been included, leading to significant changes in ATP concentration. Under rapid release, the concentration at the surface increased relative to the bulk concentration.

A Statistical Method to Evaluate the Effect of Shear Stress on Endothelial Gene Expression in vivo

Objective To determine the effect of hemodynamic shear stress on endothelial gene expression in vivo. Methods Computational fluid dynamics (CFD) simulations were performed in three realistic models of the porcine abdominal aorta and its iliac artery branches. Using these calculations, a shear stress probability map in the iliac arteries was generated. This map showed the likelihood that a particular location would experience high, medium, or low time average shear stress. Based on this map, three anatomical regions likely to experience high, medium, and low shear stress levels were identified. The median time-average shear stress in these regions were 32.1, 15.2, and 9.7 dyn/cm2, respectively. Subsequently, endothelial cell RNA was collected from these regions in five additional porcine iliac arteries. Gene expression of select genes was measured in these samples using quantitative real time PCR. Results for the high and low shear regions are reported relative to the medium shear region. Results Expression of the genes tubulin, beta-catenin, c-jun, MCP-1, and VCAM was elevated in both the low and high shear stress regions relative to the medium shear region. ICAM expression was downregulated in both the low and high shear regions relative to the medium shear region. These effects were significant in nearly all cases. Conclusions This statistical method permits evaluation of phenotypic differences that arise due to hemodynamic forces in in vivo specimens. Arterial regions subjected to distinct levels of time average shear stress exhibit differences in endothelial gene expression. Supported by NIH grant H50442.

Alteration of Reynolds number and flow partition modify wall shear stresses at arterial branches in a way that can explain age- and species-dependent patterns of arterial disease

It is assumed that critical hemodynamic factors play an important role in the onset, localization and degree of post-operative complications, for example, thrombosis and restenosis. Of special interest are sudden expansion flows, which may occur in straight artery segments such as the common carotid after endarterectomy or end-to-end anastomoses. Sudden expansion geometries are possible origins of early post-operative emboli and significant myointimal hyperplasia resulting in early or late complications. Transient laminar axisymmetric and fully three-dimensional blood flows were simulated employing a validated finite volume code in conjunction with a Runge–Kutta particle tracking technique. Disturbed flow indicators, which may predict the onset of thrombosis and/or restenosis, were identified and employed to evaluate 90°-step and smooth expansion geometries.Smooth expansion geometries have weaker disturbed flow features than step expansion geometries. Specifically, the regions near the expansion wall and the reattachment point are susceptible to both atherosclerotic lesion and thrombi formations as indicated by non-uniform hemodynamic indicators such as near-zero wall shear stress and elevated wall shear stress gradients as well as blood particle accumulation and deposition. A new parameter, the wall shear stress angle deviation (WSSAD) has been introduced, which indicates areas of abnormal endothelial cell morphology and particle wall deposition. In turn, regions of low wall shear stress and high wall shear stress gradients are recognized as susceptible sites for arterial diseases. Thus, it is interesting to note that high WSSAD surface areas cover low wall shear stress, high wall shear stress gradient locations as well as high wall particle deposition.A gradual change in step expansion geometry provides better results in terms of WSSAD values and hence potentially reducing atherosclerosis as well as thrombi formation.

Double Jeopardy

See related article, pages 149–156 In this issue of Circulation Research , Woollard et al identify a critical role of pathologically elevated levels of soluble P-selectin (sP-selectin) found in the plasma of patients with peripheral arterial occlusive disease (PAOD) that promotes the activation of neutrophils and induces their adhesion to fibrinogen and platelet monolayers.1 P-selectin is an inflammatory adhesion molecule expressed on activated platelets and endothelial cells.2 During the last two decades, the role of inflammatory cells and adhesion molecules in the development and progression of vascular diseases has become apparent, and it is now recognized that many of the cellular and molecular events that underlie atherosclerotic vascular disease are inflammatory in nature.3,4 One manifestation of atherosclerosis is PAOD, which is characterized by intermittent claudication and critical limb ischemia.5 The first case of intermittent claudication caused by arterial occlusive disease was described as early as 1831 by Jean Bouley, and a few years later Heinrich Erb proposed walking as a therapy for intermittent claudication6; however, the precise mechanism of development of PAOD as well as the optimal treatments of this disease still remain to be determined. Numerous studies have identified soluble inflammatory markers including acute phase proteins and adhesion molecules as risk factors for cardiovascular disease, but it is unclear whether they play causative roles as active participants in the initiation and maintenance of the inflammation (mediators) or whether they merely indicate the existence of the disease (markers). Soluble P-selectin is elevated in patients with vascular disorders such as hypertension,7 hypercholesterolaemia, …

Shear stress affects the intracellular distribution of eNOS: direct demonstration by a novel in vivo technique

The focal location of atherosclerosis in the vascular tree is correlated with local variations in shear stress. We developed a method to induce defined variations in shear stress in a straight vessel segment of a mouse. To this end, a cylinder with a tapered lumen was placed around the carotid artery, inducing a high shear stress field. Concomitantly, regions of low shear stress and oscillatory shear stress were created upstream and down-stream of the device, respectively. This device was used in mice transgenic for an eNOS3GFP fusion gene. We observed a strong induction of endothelial nitric oxide synthase-green fluorescent protein (eNOS-GFP) mRNA expression in the high shear stress region compared with the other regions (P < .05). Quantification of eNOS-GFP fluorescence or of immunoreactivity to the Golgi complex or to platelet endothelial cell adhesion molecule 1 (PECAM-1) showed an increase in the high shear stress region (P < .05) compared with nontreated carotid arteries. Colocalization of eNOS-GFP with either the Golgi complex or PECAM-1 also responded to alterations of shear stress. In conclusion, we showed a direct response of mRNA and protein expression in vivo to induced variations of shear stress. This model provides the opportunity to study the relationship between shear stress alterations, gene expression, and atherosclerosis.

Multiscale Analysis of Surface Thrombosis In Vivo in a Left Ventricular Assist System

Thrombosis limits the success of ventricular assist devices as the demand for alternatives to heart transplants is increasing. This study mapped the occurrence of thrombosis in a left ventricular assist system (LVAS) to better understand the biologic response to these devices. Nine calves divided into two groups were implanted with LVAS for 28 to 30 days. One group was anticoagulated, whereas the second group received no long-term anticoagulation. The blood-contacting poly(urethane urea) surfaces of blood sacs in the LVAS were examined for macroscopic thrombi upon retrieval. The sac was partitioned into eight sections and imaged for thrombi by scanning electron microscopy. No difference in thrombosis was observed macroscopically between the groups. Anticoagulation appeared to result in reduction of platelet-like structures, but the presence of fibrin-like structures remained similar between groups. Regional differences correlating with high and low shear stress regions were observed. At the macroscale, fewer thrombi were recorded in the high shear stress ports. At the microscale, features resembling fibrin were observed primarily in the ports and platelet-like features were common in lower shear stress regions. These variations in thrombosis with anticoagulation and location are likely due to varied fluid dynamics within the LVAS blood sac.

WHOLE FLOWFIELD PARTICLE IMAGE VELOCIMETRY MEASUREMENTS IN A 10 CC PULSATILE PEDIATRIC ASSIST PUMP

The analysis of a whole flowfield particle image velocimetry study of the 10 cc Penn State pulsatile pediatric ventricular assist device (PVAD) is presented. The pressures and flow through the PVAD are controlled with a mock circulatory loop. The blood analog (Sodium Iodide/Xanthan gum) solution is a good match for the viscoelastic properties of blood across the shear rate range. An earlier version of the Penn State PVAD has shown a much higher incidence of clot formation than does the adult pump from which it was derived. Therefore, a focus of the current study is on the wall shear stress. We identify regions of low wall shear stress throughout the pump cycle and suggest how the pump design might be geometrically modified to increase the flow rate there. In addition, we analyze the flow in the valve regions, as predicting the result of downsizing the valves from the adult pump is not straightforward.

Letter Regarding Article by Korshunov and Berk, “Strain-Dependent Vascular Remodeling: The ‘Glagov Phenomenon’ Is Genetically Determined”

To the Editor: Drs Korshunov and Berk1 described the modulation of vascular remodeling in response to a sudden decrease of shear stress induced by partial ligation of the carotid artery of different mouse strains. Although a similar study was previously conducted with similar mouse strains in which the carotid artery was totally occluded,2 Korshunov and Berk argued that their partial ligation model is more relevant to human carotid intima-media thickness and coronary atherosclerosis and concluded from their measurements that the Glagov phenomenon has a genetic background. To strengthen the relevance of their data, the authors compared their results—outward remodeling under low flow conditions—to our data in human atherosclerotic coronary arteries.3 Indeed, we found that in segments with lumen preservation because of outward vascular remodeling, atherosclerotic plaques were observed at relatively low shear stress regions.3 Because vascular remodeling is defined as an average over the cross …

Cell docking inside microwells within reversibly sealed microfluidic channels for fabricating multiphenotype cell arrays

We present a soft lithographic method to fabricate multiphenotype cell arrays by capturing cells within an array of reversibly sealed microfluidic channels. The technique uses reversible sealing of elastomeric polydimethylsiloxane (PDMS) molds on surfaces to sequentially deliver various fluids or cells onto specific locations on a substrate. Microwells on the substrate were used to capture and immobilize cells within low shear stress regions inside channels. By using an array of channels it was possible to deposit multiple cell types, such as hepatocytes, fibroblasts, and embryonic stem cells, on the substrates. Upon formation of the cell arrays on the substrate, the PDMS mold could be removed, generating a multiphenotype array of cells. In addition, the orthogonal alignment and subsequent attachment of a secondary array of channels on the patterned substrates could be used to deliver fluids to the patterned cells. The ability to position many cell types on particular regions within a two dimensional substrate could potentially lead to improved high-throughput methods applicable to drug screening and tissue engineering.