Wall Shear Stress Conditions Research Articles

Investigation of Scaling and Materials’ Performance in Simulated Geothermal Brine

Geothermal energy generation faces challenges in efficiency, partly due to restrictions on reinjection temperatures caused by scaling issues. Therefore, developing strategies to prevent scaling is critical. This study aims to simulate the scaling tendencies and corrosion effects of geothermal fluids on various construction materials used in scaling reactor/retention tank systems. A range of materials, including carbon steel, austenitic stainless steel, duplex stainless steel, two proprietary two-part epoxy coatings, and thermally sprayed aluminium (TSA), were tested in a simulated geothermal brine. Experiments were conducted in a laboratory vessel designed to replicate the wall shear stress conditions expected in a scaling reactor. The tests revealed varying scaling tendencies among the materials, with minimal corrosion observed. The dominant scale formed was calcium carbonate, consistent with geochemical modelling. The findings suggest that despite the high operating temperatures, the risk of corrosion remains low due to the brine’s low chloride content, while the wettability of materials after immersion may serve as a useful indicator for selecting those that promote scaling.

A 96-wells fluidic system for high-throughput screenings under laminar high wall shear stress conditions

The ability of endothelial cells to respond to blood flow is fundamental for the correct formation and maintenance of a functional and hierarchically organized vascular network. Defective flow responses, in particular related to high flow conditions, have been associated with atherosclerosis, stroke, arteriovenous malformations, and neurodegenerative diseases. Yet, the molecular mechanisms involved in high flow response are still poorly understood. Here, we described the development and validation of a 96-wells fluidic system, with interchangeable cell culture and fluidics, to perform high-throughput screenings under laminar high-flow conditions. We demonstrated that endothelial cells in our newly developed 96-wells fluidic system respond to fluid flow-induced shear stress by aligning along the flow direction and increasing the levels of KLF2 and KLF4. We further demonstrate that our 96-wells fluidic system allows for efficient gene knock-down compatible with automated liquid handling for high-throughput screening platforms. Overall, we propose that this modular 96-well fluidic system is an excellent platform to perform genome-wide and/or drug screenings to identify the molecular mechanisms involved in the responses of endothelial cells to high wall shear stress.

P19: In vitro Evaluation of Platelet Damage of Turbulent and Laminar Flow at Same Wall Shear Stress Conditions

P19: In vitro Evaluation of Platelet Damage of Turbulent and Laminar Flow at Same Wall Shear Stress Conditions

A Compressed Collagen Construct for Studying Endothelial-Smooth Muscle Cell Interaction Under High Shear Stress.

The coculture of vascular endothelial cells (ECs) on collagen gels containing smooth muscle cells (SMCs) has been carried out to investigate cellular interactions associated with blood vessel pathophysiology under wall shear stress (WSS) conditions. However, due to a lack of gel stiffness, the previous collagen gel coculture constructs are difficult to use for pathologic higher WSS conditions. Here, we newly constructed a coculture model with centrifugally compressed cell-collagen combined construct (C6), which withstands higher WSS conditions. The elastic modulus of C6 was approximately 6 times higher than that of the uncompressed collagen construct. The level of α-smooth muscle actin, a contractile SMC phenotype marker observed in healthy arteries, was elevated in C6 compared with that of the uncompressed construct, and further increased by exposure to a physiological level WSS of 2 Pa, but not by a pathological level of 20 Pa. WSS conditions of 2 and 20 Pa also induced different expression ratios of matrix metalloproteinases and their inhibitors in the C6 coculture model but did not in monocultured ECs and SMCs. The C6 coculture model will be a powerful tool to investigate interactions between ECs and SMCs under pathologically high WSS conditions.

Multiscale Modeling of Vascular Remodeling Induced by Wall Shear Stress

ObjectiveHemodynamics-induced low wall shear stress (WSS) is one of the critical reasons leading to vascular remodeling. However, the coupling effects of WSS and cellular kinetics have not been clearly modeled. The aim of this study was to establish a multiscale modeling approach to reveal the vascular remodeling behavior under the interaction between the macroscale of WSS loading and the microscale of cell evolution.MethodsComputational fluid dynamics (CFD) method and agent-based model (ABM), which have significantly different characteristics in temporal and spatial scales, were adopted to establish the multiscale model. The CFD method is for the second/organ scale, and the ABM is for the month/cell scale. The CFD method was used to simulate blood flow in a vessel and obtain the WSS in a vessel cross-section. The simulations of the smooth muscle cell (SMC) proliferation/apoptosis and extracellular matrix (ECM) generation/degradation in a vessel cross-section were performed by using ABM. During the simulation of the vascular remodeling procedure, the damage index of the SMC and ECM was defined as deviation from the obtained WSS. The damage index decreased gradually to mimic the recovery of WSS-induced vessel damage.Results(1) The significant wall thickening region was consistent with the low WSS region. (2) There was no evident change of wall thickness in the normal WSS region. (3) When the damage index approached to 0, the amount and distribution of SMCs and ECM achieved a stable state, and the vessel reached vascular homeostasis.ConclusionThe established multiscale model can be used to simulate the vascular remodeling behavior over time under various WSS conditions.

Endothelial Cell Distribution After Flow Exposure With Two Stent Struts Placed in Different Angles.

Stent implantation has been a primary treatment for stenosis and other intravascular diseases. However, the struts expansion procedure might cause endothelium lesion and the structure of the struts could disturb the blood flow environment near the wall of the blood vessel. These changes could damage the vascular innermost endothelial cell (EC) layer and pose risks of restenosis and post-deployment thrombosis. This research aims to investigate the effect of flow alterations on EC distribution in the presence of gap between two struts within the parallel flow chamber. To study how the gap presence impacts EC migration and the endothelialization effect on the surface of the struts, two struts were placed with specific orientations and positions on the EC layer in the flow chamber. After a 24-h exposure under wall shear stress (WSS), we observed the EC distribution conditons especially in the gap area. We also conducted computational fluid dynamics (CFD) simulations to calculate the WSS distribution. High EC-concentration areas on the bottom plate corresponded to the high WSS by the presence of gap between the two struts. To find the relation between the WSS and EC distributions on the fluorescence images, WSS condition by CFD simulation could be helpful for the EC distribution. The endothelialization rate, represented by EC density, on the downstream sides of both struts was higher than that on the upstream sides. These observations were made in the flow recirculation at the gap area between two struts. On two side surfaces between the gaps, meaning the downstream at the first and the upstream at the second struts, EC density differences on the downstream surfaces of the first strut were higher than on the upstream surfaces of the second strut. Finally, EC density varied along the struts when the struts were placed at tilted angles. These results indicate that, by the presence of gap between the struts, ECs distribution could be predicted in both perpendicular and tiled positions. And tiled placement affect ECs distribution on the strut side surfaces.

Smooth muscle cells affect differential nanoparticle accumulation in disturbed blood flow-induced murine atherosclerosis.

Atherosclerosis is a lipid-driven chronic inflammatory disease that leads to the formation of plaques in the inner lining of arteries. Plaques form over a range of phenotypes, the most severe of which is vulnerable to rupture and causes most of the clinically significant events. In this study, we evaluated the efficacy of nanoparticles (NPs) to differentiate between two plaque phenotypes based on accumulation kinetics in a mouse model of atherosclerosis. This model uses a perivascular cuff to induce two regions of disturbed wall shear stress (WSS) on the inner lining of the instrumented artery, low (upstream) and multidirectional (downstream), which, in turn, cause the development of an unstable and stable plaque phenotype, respectively. To evaluate the influence of each WSS condition, in addition to the final plaque phenotype, in determining NP uptake, mice were injected with NPs at intermediate and fully developed stages of plaque growth. The kinetics of artery wall uptake were assessed in vivo using dynamic contrast-enhanced magnetic resonance imaging. At the intermediate stage, there was no difference in NP uptake between the two WSS conditions, although both were different from the control arteries. At the fully-developed stage, however, NP uptake was reduced in plaques induced by low WSS, but not multidirectional WSS. Histological evaluation of plaques induced by low WSS revealed a significant inverse correlation between the presence of smooth muscle cells and NP accumulation, particularly at the plaque-lumen interface, which did not exist with other constituents (lipid and collagen) and was not present in plaques induced by multidirectional WSS. These findings demonstrate that NP accumulation can be used to differentiate between unstable and stable murine atherosclerosis, but accumulation kinetics are not directly influenced by the WSS condition. This tool could be used as a diagnostic to evaluate the efficacy of experimental therapeutics for atherosclerosis.

Effects of Low and High Aneurysmal Wall Shear Stress on Endothelial Cell Behavior: Differences and Similarities.

Background: Intracranial aneurysms (IAs) result from abnormal enlargement of the arterial lumen. IAs are mostly quiescent and asymptomatic, but their rupture leads to severe brain damage or death. As the evolution of IAs is hard to predict and intricates medical decision, it is essential to improve our understanding of their pathophysiology. Wall shear stress (WSS) is proposed to influence IA growth and rupture. In this study, we investigated the effects of low and supra-high aneurysmal WSS on endothelial cells (ECs).Methods: Porcine arterial ECs were exposed for 48 h to defined levels of shear stress (2, 30, or 80 dyne/cm2) using an Ibidi flow apparatus. Immunostaining for CD31 or γ-cytoplasmic actin was performed to outline cell borders or to determine cell architecture. Geometry measurements (cell orientation, area, circularity and aspect ratio) were performed on confocal microscopy images. mRNA was extracted for RNAseq analysis.Results: ECs exposed to low or supra-high aneurysmal WSS were more circular and had a lower aspect ratio than cells exposed to physiological flow. Furthermore, they lost the alignment in the direction of flow observed under physiological conditions. The effects of low WSS on differential gene expression were stronger than those of supra-high WSS. Gene set enrichment analysis highlighted that extracellular matrix proteins, cytoskeletal proteins and more particularly the actin protein family were among the protein classes the most affected by shear stress. Interestingly, most genes showed an opposite regulation under both types of aneurysmal WSS. Immunostainings for γ-cytoplasmic actin suggested a different organization of this cytoskeletal protein between ECs exposed to physiological and both types of aneurysmal WSS.Conclusion: Under both aneurysmal low and supra-high WSS the typical arterial EC morphology molds to a more spherical shape. Whereas low WSS down-regulates the expression of cytoskeletal-related proteins and up-regulates extracellular matrix proteins, supra-high WSS induces opposite changes in gene expression of these protein classes. The differential regulation in EC gene expression observed under various WSS translate into a different organization of the ECs’ architecture. This adaptation of ECs to different aneurysmal WSS conditions may affect vascular remodeling in IAs.

Influence of Ceramic Membrane Surface Characteristics on the Flux Behavior of a Complex Fermentation Broth.

The valorization of agro-industrial residues using yeasts as biocatalysts requires efficient methods for biomass separation. Filtration with ceramic membranes is suitable for this task, however, the challenge of flux decline and the unavoidable cleaning must be taken into account. We investigated the filtration of fermentation broth and its components using tubular microfiltration and ultrafiltration membranes, and hollow-fiber ultrafiltration membranes, with cut-offs of 30 and 200 nm. The steady-state flux was limited by fouling under comparable wall shear stress conditions but increased when the wall shear stress was higher. Single-component filtration with two 30 nm tubular ultrafiltration membranes, whose average surface roughness ranged from 1.0 to 3.9 µm, showed that smoother surfaces experience less biomass fouling under more intense hydrodynamic conditions. Furthermore, we showed experimentally and by scanning electron microscopy in filtration with 30 nm tubular membranes that the thickness of the first separation layer is responsible for the degree of irreversible resistance caused by the deposition of organic material in the membrane pores. The thickness of this layer should therefore be minimized without compromising mechanical stability.

Comparative Assessment of Tubular Ceramic, Spiral Wound, and Hollow Fiber Membrane Microfiltration Module Systems for Milk Protein Fractionation.

The fractionation efficiency of hollow fiber membranes (HFM) for milk protein fractionation was compared to ceramic tubular membranes (CTM) and spiral wound membranes (SWM). HFM combine the features of high membrane packing density of SWM and the more defined flow conditions and better control of membrane fouling in the open flow channel cross-sections of CTM. The aim was to comparatively analyze the effect of variations in local pressure and flow conditions while using single industrially sized standard modules with similar dimensions and module footprints (module diameter and length). The comparative assessment with varied transmembrane pressure was first applied for a constant feed volume flow rate of 20 m3 h−1 and, secondly, with the same axial pressure drop along the modules of 1.3 bar m−1, similar to commonly applied crossflow velocity and wall shear stress conditions at the industrial level. Flux, transmission factor of proteins (whey proteins and serum caseins), and specific protein mass flow per area membrane and per volume of module installed were determined as the evaluation criteria. The casein-to-whey protein ratios were calculated as a measure for protein fractionation effect. Results obtained show that HFM, which so far are under-represented as standard module types in industrial dairy applications, appear to be a competitive alternative to SWM and CTM for milk protein fractionation.

Fluid-dynamics and biological features of unstable plaques: different shear stress for different plaques

Abstract Background The use of Optical Coherence Tomography (OCT) in acute coronary syndromes (ACS) allows recognizing ruptured fibrous cap (RFC) and intact fibrous cap (IFC) culprit lesions. The biological differences between them, as recently pointed out in translation studies, highlight different mechanisms for a similar clinical manifestation that might deserve different therapeutic approaches. The relationship between endothelial wall shear stress (WSS) and ACS has been demonstrated, however the differences in WSS features between RFC and IFC have not been elucidated. Purpose The aim of this study is to provide a fluid-dynamic and biological description of unstable and stable (SA) plaques, according to OCT analysis. Methods We enrolled 10 SA and 20 Non-ST Elevation Myocardial Infarction (NSTEMI)-ACS patients, with IFC (n=10) and RFC (n=10) culprit lesions according to OCT analysis. We performed Real-time PCR primer array on pooled Peripheral Blood Mononuclear Cell (PBMC) for 30 different molecules whose expression is strictly dependent on WSS. High-fidelity 3D-coronary artery models were created for 3 patients per group, applying previously validated methodologies. Results Among the groups we found a broad difference in molecular expression (Fig. 1A), with RFC displaying higher levels of molecules involved in vasoconstriction/dilatation (EDN1, NOS3), cellular adhesion (ICAM1), and peptidase inhibition (PI16). A significantly higher WSS was observed in RFC group (p<0.001, Fig 1B and C), with larger variability and larger areas exposed to both low and high WSS (Fig. 1D). Interestingly, the molecules overexpressed in RFC are known to be upregulated by high WSS. Conclusions Our data demonstrated that IFC and RFC unstable plaques are associated with different WSS conditions, alongside with the expression of different molecular patterns specifically related to altered WSS. In the era of precision medicine these findings may have relevant therapeutical implications. Funding Acknowledgement Type of funding source: None

3D Perfusable Hydrogel Recapitulating the Cancer Dynamic Environment to in Vitro Investigate Metastatic Colonization.

Metastasis is a dynamic process involving the dissemination of circulating tumor cells (CTCs) through blood flow to distant tissues within the body. Nevertheless, the development of an in vitro platform that dissects the crucial steps of metastatic cascade still remains a challenge. We here developed an in vitro model of extravasation composed of (i) a single channel-based 3D cell laden hydrogel representative of the metastatic site, (ii) a circulation system recapitulating the bloodstream where CTCs can flow. Two polymers (i.e., fibrin and alginate) were tested and compared in terms of mechanical and biochemical proprieties. Computational fluid-dynamic (CFD) simulations were also performed to predict the fluid dynamics within the polymeric matrix and, consequently, the optimal culture conditions. Next, once the platform was validated through perfusion tests by fluidically connecting the hydrogels with the external circuit, highly metastatic breast cancer cells (MDA-MB-231) were injected and exposed to physiological wall shear stress (WSS) conditions (5 Dyn/cm2) to assess their migration toward the hydrogel. Results indicated that CTCs arrested and colonized the polymeric matrix, showing that this platform can be an effective fluidic system to model the first steps occurring during the metastatic cascade as well as a potential tool to in vitro elucidate the contribution of hemodynamics on cancer dissemination to a secondary site.

Effect of high wall shear stress condition on the MMP expressions in an EC-SMC coculture model

Effect of high wall shear stress condition on the MMP expressions in an EC-SMC coculture model

Homeobox B9 integrates bone morphogenic protein 4 with inflammation at atheroprone sites.

AimsAtherosclerosis develops near branches and bends of arteries that are exposed to disturbed blood flow which exerts low wall shear stress (WSS). These mechanical conditions alter endothelial cells (EC) by priming them for inflammation and by inducing turnover. Homeobox (Hox) genes are developmental genes involved in the patterning of embryos along their anterior–posterior and proximal–distal axes. Here we identified Hox genes that are regulated by WSS and investigated their functions in adult arteries.Methods and resultsEC were isolated from inner (low WSS) and outer (high WSS) regions of the porcine aorta and the expression of Hox genes was analysed by quantitative real-time PCR. Several Hox genes (HoxA10, HoxB4, HoxB7, HoxB9, HoxD8, HoxD9) were significantly enriched at the low WSS compared to the high WSS region. Similarly, studies of cultured human umbilical vein EC (HUVEC) or porcine aortic EC revealed that the expression of multiple Hox genes (HoxA10, HoxB9, HoxD8, HoxD9) was enhanced under low (4 dyn/cm2) compared to high (13 dyn/cm2) WSS conditions. Gene silencing studies identified Hox genes (HoxB9, HoxD8, HoxD9) that are positive regulators of inflammatory molecule expression in EC exposed to low WSS, and others (HoxB9, HoxB7, HoxB4) that regulated EC turnover. We subsequently focused on HoxB9 because it was strongly up-regulated by low WSS and, uniquely, was a driver of both inflammation and proliferation. At a mechanistic level, we demonstrate using cultured EC and murine models that bone morphogenic protein 4 (BMP4) is an upstream regulator of HoxB9 which elicits inflammation via induction of numerous inflammatory mediators including TNF and downstream NF-κB activation. Moreover, the BMP4-HoxB9-TNF pathway was potentiated by hypercholesterolaemic conditions.ConclusionsLow WSS induces multiple Hox genes that control the activation state and turnover of EC. Notably, low WSS activates a BMP4-HoxB9-TNF signalling pathway to initiate focal arterial inflammation, thereby demonstrating integration of the BMP and Hox systems in vascular pathophysiology.

The optimal incubation time for in vitro hemocompatibility testing: Assessment using polymer reference materials under pulsatile flow with physiological wall shear stress conditions.

During hemocompatibility testing, activation products may reach plateau values which can result in less distinction between hemocompatible and hemo‐incompatible materials. Of concern is an underestimation of the blood activation caused by the biomaterial of interest, which may result in a false assessment of hemocompatibility. To elucidate the optimal incubation time for in vitro hemocompatibility testing, we used the Haemobile circulation model with human whole blood. Blood from healthy volunteers was in vitro incubated under pulsatile flow with physiological wall shear stress conditions at 37°C for 30, 60, 120, or 240 min. Test loops containing low‐density polyethylene and polydimethylsiloxane served as low and high reference materials, that is, hemocompatible and hemo‐incompatible biomaterials, respectively. In addition, empty loops served as a negative reference. Thrombogenicity, platelet function, inflammatory response, coagulation, and hemolysis between references and incubation times were compared. We found that thrombogenicity and platelet function were significantly affected by both the duration of incubation and the type of material. In particular, thrombogenicity and platelet function assessments were affected by incubation time. We found that an exposure time of 60 min was sufficient, and for almost all variables an optimal incubation time to discriminate between the low and high reference material. © 2019 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2335–2342, 2019.

Disturbed Blood Flow induces Arterial Stiffening Through Thrombospondin‐1

BACKGROUNDArterial stiffness and wall shear stress are powerful determinants of cardiovascular health, and arterial stiffness is associated with increased cardiovascular mortality. Low and oscillatory wall shear stress, termed disturbed flow (d‐flow), promotes atherosclerotic arterial remodeling, but the relationship between d‐flow and arterial stiffness is not well understood. The objective of this study was to define the role of d‐flow on arterial stiffening and discover the relevant signaling pathways by which d‐flow stiffens arteries.METHODSD‐flow was induced in the carotid arteries of young and old mice of both sexes. Arterial stiffness was quantified ex vivo with cylindrical biaxial mechanical testing and in vivo from duplex ultrasound and compared with unmanipulated carotid arteries from 80‐week‐old mice. Gene expression and pathway analysis was performed on endothelial cell‐enriched RNA and validated by immunohistochemistry. In vitro testing of signaling pathways was performed under oscillatory and laminar wall shear stress conditions. Human arteries from regions of d‐flow and stable flow were tested ex vivo to validate critical results from the animal model.RESULTSD‐flow induced arterial stiffening through collagen deposition after partial carotid ligation, and the degree of stiffening was similar to that of unmanipulated carotid arteries from 80‐week‐old mice. Intimal gene pathway analyses identified transforming growth factor‐β pathways as having a prominent role in this stiffened arterial response, but this was attributable to thrombospondin‐1 (TSP‐1) stimulation of profibrotic genes and not changes to transforming growth factor‐β. In vitro and in vivo testing under d‐flow conditions identified a possible role for TSP‐1 activation of transforming growth factor‐β in the upregulation of these genes. TSP‐1 knockout animals had significantly less arterial stiffening in response to d‐flow than wild‐type carotid arteries. Human arteries exposed to d‐flow had similar increases TSP‐1 and collagen gene expression as seen in our model.CONCLUSIONSTSP‐1 has a critical role in shear‐mediated arterial stiffening that is mediated in part through TSP‐1's activation of the profibrotic signaling pathways of transforming growth factor‐β. Molecular targets in this pathway may lead to novel therapies to limit arterial stiffening and the progression of disease in arteries exposed to d‐flow.Support or Funding InformationFunding for this work was provided by the National Institutes of Health, KO8HL119592 (to Dr Brewster), HL119798 and HL124879 (to Dr Jo); the Society for Vascular Surgery/American College of Surgeons Mentored Clinical Scientist Research Career Development Award (to Dr Brewster); the American Heart Association, IRG1470001 (to Dr Brewster); departmental support from the Emory University Department of Surgery (Dr John F. Sweeney) (to Dr Brewster); and the John and Jan Portman Professorship (to Dr Jo) from Emory University School of Medicine and Georgia Institute of Technology.This study was supported in part by the Animal Physiology Core, which is subsidized by Emory University and Children's Healthcare of Atlanta. Additional support was provided by the Office of the Director of the National Institutes of Health under Award Number S10OD021748. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the National Institutes of Health.

Response Surface based Analysis to Improve the Intravascular Stent Design Performance on Optimizing Wall Shear Stress Conditions

Response Surface based Analysis to Improve the Intravascular Stent Design Performance on Optimizing Wall Shear Stress Conditions

Disturbed Flow Promotes Arterial Stiffening Through Thrombospondin-1.

Arterial stiffness and wall shear stress are powerful determinants of cardiovascular health, and arterial stiffness is associated with increased cardiovascular mortality. Low and oscillatory wall shear stress, termed disturbed flow (d-flow), promotes atherosclerotic arterial remodeling, but the relationship between d-flow and arterial stiffness is not well understood. The objective of this study was to define the role of d-flow on arterial stiffening and discover the relevant signaling pathways by which d-flow stiffens arteries. D-flow was induced in the carotid arteries of young and old mice of both sexes. Arterial stiffness was quantified ex vivo with cylindrical biaxial mechanical testing and in vivo from duplex ultrasound and compared with unmanipulated carotid arteries from 80-week-old mice. Gene expression and pathway analysis was performed on endothelial cell-enriched RNA and validated by immunohistochemistry. In vitro testing of signaling pathways was performed under oscillatory and laminar wall shear stress conditions. Human arteries from regions of d-flow and stable flow were tested ex vivo to validate critical results from the animal model. D-flow induced arterial stiffening through collagen deposition after partial carotid ligation, and the degree of stiffening was similar to that of unmanipulated carotid arteries from 80-week-old mice. Intimal gene pathway analyses identified transforming growth factor-β pathways as having a prominent role in this stiffened arterial response, but this was attributable to thrombospondin-1 (TSP-1) stimulation of profibrotic genes and not changes to transforming growth factor-β. In vitro and in vivo testing under d-flow conditions identified a possible role for TSP-1 activation of transforming growth factor-β in the upregulation of these genes. TSP-1 knockout animals had significantly less arterial stiffening in response to d-flow than wild-type carotid arteries. Human arteries exposed to d-flow had similar increases TSP-1 and collagen gene expression as seen in our model. TSP-1 has a critical role in shear-mediated arterial stiffening that is mediated in part through TSP-1's activation of the profibrotic signaling pathways of transforming growth factor-β. Molecular targets in this pathway may lead to novel therapies to limit arterial stiffening and the progression of disease in arteries exposed to d-flow.

How neutrophils resist shear stress at blood vessel walls: molecular mechanisms, subcellular structures, and cell-cell interactions.

Neutrophils are the first cells arriving at sites of tissue injury or infection to combat invading pathogens. Successful neutrophil recruitment to sites of inflammation highly depends on specific molecular mechanisms, fine-tuning the received information into signaling pathways and converting them into well-described recruitment steps. This review highlights the impact of vascular flow conditions on neutrophil recruitment and the multitude of mechanisms developed to enable this sophisticated process under wall shear stress conditions. The recruitment process underlies a complex interplay between adhesion and signaling molecules, as well as chemokines, in which neutrophils developed specific mechanisms to travel to sites of lesion in low and high shear stress conditions. Rolling, as the first step in the recruitment process, highly depends on endothelial selectins and their ligands on neutrophils, inducting of intracellular signaling and subsequently activating β2 integrins, enabling adhesion and postadhesion events. In addition, subcellular structures, such as microvilli, tethers, and slings allow the cell to arrest, even under high wall shear stress. Thereby, microvilli that are pulled out from the cell body form tethers that develop into slings upon their detachment from the substrate. In addition to the above-described primary capture, secondary capture of neutrophils via neutrophil-neutrophil or neutrophil-platelet interaction promotes the process of neutrophil recruitment to sites of lesion. Thus, precise mechanisms based on a complex molecular interplay, subcellular structures, and cell-cell interactions turn the delicate process of neutrophil trafficking during flow into a robust response allowing effective neutrophil accumulation at sites of injury.

Facile endothelium protection from TNF-α inflammatory insult with surface topography.

Adverse events triggered by the direct contact between blood and synthetic materials constitute a sincere shortcoming of cardiovascular implant technology. A well-connected autologous endothelium, generated through the process of endothelialization, impedes such interaction and endows the implant luminal interface with optimal protection. The endothelialization of artificial substrates is the result of a complex interplay between endothelial cells (ECs), surface topography, and flow-generated wall shear stress (WSS). This is however tainted by the pro-inflammatory signaling, typical of cardiovascular patients, which compromises endothelial integrity and survival. Here, we challenge human endothelial monolayers with the pro-inflammatory factor TNF-α under realistic WSS conditions. In these experimental settings we demonstrate that the simple contact between ECs and an optimized surface geometry can inhibit NF-kB activation downstream of TNF-α yielding increased stability of VE-Cadherin mediated cell-to-cell junctions and of focal adhesions. Therefore the here-presented topographic modification can be implemented on a range of artificial substrates enabling their endothelialization under supra-physiological flow and in the presence of pro-inflammatory insults. These new findings constitute an important step toward achieving the full hemocompatibility of cardiovascular implants.