Effect Of Erythrocyte Aggregation Research Articles

Axial shear rate: A hemorheological factor for erythrocyte aggregation under Womersley flow in an elastic vessel based on numerical simulation

Erythrocyte aggregation (EA) is a highly dynamic, vital phenomenon to interpreting human hemorheology, which would be helpful for the diagnosis and prediction of circulatory anomalies. Previous studies of EA on erythrocyte migration and the Fåhraeus Effect are based on the microvasculature. They have not considered the natural pulsatility of the blood flow or large vessels and mainly focused on shear rate along radial direction under steady flow to comprehend the dynamic properties of EA. To our knowledge, the rheological characteristics of non-Newtonian fluids under Womersley flow have not reflected the spatiotemporal behaviors of EA or the distribution of erythrocyte dynamics (ED). Hence, it needs to interpret the ED affected by temporal and spatial flow variation to understand the effect of EA under Womersley flow. Here, we demonstrated the numerically simulated ED to decipher EA's rheological role in axial shear rate under Womersley flow. In the present study, the temporal and spatial variations of the local EA were found to mainly depend on the axial shear rate under Womersley flow in an elastic vessel, while mean EA decreased with radial shear rate. The localized distribution of parabolic or M-shape clustered EA was found in a range of the axial shear rate profile (−15 to 15s−1) at low radial shear rates during a pulsatile cycle. However, the linear formation of rouleaux was realized without local clusters in a rigid wall where the axial shear rate is zero. In vivo, the axial shear rate is usually considered insignificant, especially in straight arteries, but it has a great impact on the disturbed blood flow due to the geometrical properties, such as bifurcations, stenosis, aneurysm, and the cyclic variation of pressure. Our findings regarding axial shear rate provide new insight into the local dynamic distribution of EA, which is a critical player in blood viscosity. These will provide a basis for the computer-aided diagnosis of hemodynamic-based cardiovascular diseases by decreasing the uncertainty in the pulsatile flow calculation.

Surface tension driven flow of blood in a rectangular microfluidic channel: Effect of erythrocyte aggregation

Microfluidic platforms have increasingly been explored for in vitro blood diagnostics and for studying complex microvascular processes. The perfusion of blood in such devices is typically achieved through pressure-driven setups. Surface tension driven blood flow provides an alternative flow delivery option, and various studies in the literature have examined the behavior of blood flow in such fluidic devices. In such flows, the influence of red blood cell (RBC) aggregation, the phenomenon majorly responsible for the non-Newtonian nature of blood, requires particular attention. In the present work, we examine differences in the surface tension driven flow of aggregating and non-aggregating RBC and Newtonian suspensions, in a rectangular microchannel. The velocity fields were obtained using micro-PIV techniques. The analytical solution for blood velocity in the channel is developed utilizing the power law model for blood viscosity. The results showed that RBC aggregation has an impact at the late stages of the flow, observed mainly in the bluntness of the velocity profiles. At the initial stages of the flow, the shearing conditions are found moderately elevated, preventing intense RBC aggregate formation. As the flow decelerates in the channel, RBC aggregation increases, affecting the flow characteristics.

An optofluidic point-of-care device for quantitative investigation of erythrocyte aggregation during coagulation

Coagulation, the process leading to clot formation with the interplay of blood constituents, is a self-regulating mechanism, requiring attentive and periodic monitoring for numerous clinical cases. Erythrocyte aggregation (EA) is a characteristic behaviour of erythrocytes forming reversible clumps especially in vitro at low shear rates. The effect of EA during coagulation is overlooked in whole blood (WB) clotting assays, and the relationship between the two mechanisms is not well understood. We present an optofluidic point-of-care device enabling quantitative investigation of EA from 50 μl WB during the coagulation process. Not only did we explain the coagulation mechanism considering EA, but we also demonstrated coagulation time measurement from optical EA analysis. The device consists of a disposable cartridge and a handheld analyzer containing a pinch valve for fluid motion and optics for transmitted light measurement. Following the sample introduction and cessation of the valve operation, the optical signal is the lowest due to shear-induced cell disaggregation. Then, the signal increases due to EA until reaching a peak, indicating blood clotting. The working principle was proven through clinical tests for prothrombin time measurement. In addition to revealing the relation between coagulation and aggregation, this device is promising for rapid WB coagulation time measurement.

ХАРАКТЕРИСТИКА МЕТОДА АГРЕГАТОСКОПИИ ДЛЯ РЕГИСТРАЦИИ МИКРОРЕОЛОГИИ ЭРИТРОЦИТОВ

Введение. Две микрореологические характеристики определяют кровоток в системе сосудов микроциркуляции — агрегация и деформируемость эритроцитов. В подавляющем большинстве приборов для регистрации агрегации эритроцитов (АЭ) отсутствует визуализация процесса, и интерпретация данных основывается на его косвенных характеристиках. Материалы и методы. Проведено исследование АЭ на созданной установке — агрегатоскопе, получены данные регистрации картины АЭ с последующей обработкой изображения с помощью специальной программы для ЭВМ. Информативность полученных данных была проверена в сравнительных исследованиях с использованием агрегометра эритроцитов Myrenne M1 и теста СОЭ. Результаты. Получены значительные положительные корреляции (r=0,90 и r=0,86, соответственно). Показано, что агрегатоскоп дает четкую картину изменения АЭ (ее снижение) при инкубации эритроцитов с хелатором Са2+ (ЭДТА, верапамил, изобутилметилксантин, монафрам). В ответ на инкубацию с препаратами другой группы, известными как стимуляторы АЭ (CaCl2, ионофор А23187, фенилэфрин, простагландин F2α), был получен достоверный прирост агрегации. Заключение. Метод агрегатоскопии в сочетании с программной обработкой изображений является удобным и надежным инструментом оценки суспензионной стабильности крови и точным методом измерения важной микрореологической характеристики эритроцитов — их агрегации. Introduction. Aggregation and deformability of erythrocytes are two microrheological characteristics that determine the blood fl ow in microcirculation vessels. In large majority of devices for registration of erythrocyte aggregation (EA) there is no visualization of the process, and the interpretation of the data is based on its indirect characteristics. Materials and methods. EA investigation was carried out on the created unit — aggregatoscop. Data were obtained for registration of EA followed by image processing using a special computer program. Data informativeness was verified in comparative studies using erythrocyte aggregometer Myrenne M1 and erythrocyte sedimentation rate test. Results. Significant positive correlations were obtained (r=0,90 and r=0,86, respectively). It was shown that aggregatoscop gave a clear picture of EA changes (its reduction) during erythrocytes incubation with Са2+ chelator (EDTA, verapamil, isobutylmethylxanthine, monaphram). Reliable increasing of aggregation was obtained in response to incubation with other agents — EA stimulants (CaCl2, ionophore A23187, phenylephrine, prostaglandin F2α). Conclusion. Aggregoscopy method in combination with software image processing is a convenient and reliable tool for assessing of blood suspension stability and an accurate method for measuring of important microrheological characteristics of erythrocytes — their aggregation.

Quantifying local characteristics of velocity, aggregation and hematocrit of human erythrocytes in a microchannel flow.

The effect of erythrocyte aggregation on blood viscosity and microcirculatory flow is a poorly understood area of haemodynamics, especially with relevance to serious pathological conditions. Advances in microfluidics have made it possible to study the details of blood flow in the microscale, however, important issues such as the relationship between the local microstructure and local flow characteristics have not been investigated extensively. In the present study an experimental system involving simple brightfield microscopy has been successfully developed for simultaneous, time-resolved quantification of velocity fields and local aggregation of human red blood cells (RBC) in microchannels. RBCs were suspended in Dextran and phosphate buffer saline solutions for the control of aggregation. Local aggregation characteristics were investigated at bulk and local levels using statistical and edge-detection image processing techniques. A special case of aggregating flow in a microchannel, in which hematocrit gradients were present, was studied as a function of flowrate and time. The level of aggregation was found to strongly correlate with local variations in velocity in both the bulk flow and wall regions. The edge detection based analysis showed that near the side wall large aggregates are associated with regions corresponding to high local velocities and low local shear. On the contrary, in the bulk flow region large aggregates occurred in regions of low velocity and high erythrocyte concentration suggesting a combined effect of hematocrit and velocity distributions on local aggregation characteristics. The results of this study showed that using multiple methods for aggregation quantification, albeit empirical, could help towards a robust characterisation of the structural properties of the fluid.

Effect of erythrocyte aggregation at pathological levels on NO/O2 transport in small arterioles.

This study examined the effects of red blood cell (RBC) aggregation at pathological levels on NO/O2 transport in small arterioles. Transient gas diffusion simulations were performed with in vivo cell-free layer (CFL) widths data obtained from arteriolar flows in the rat cremaster muscle. The CFL data were measured at physiological and pathological levels of aggregation under reduced flow conditions (pseudoshear rate = 31.4 ± 10.5 s-1). Our results showed that the mean peak NO concentration significantly decreased with increasing the aggregation level from non-aggregating to normal-aggregating (P < 0.05) and to hyper-aggregating (P < 0.01) conditions. In contrast, the partial O2 pressure (PO2) in pathological aggregating conditions significantly increased from those under non-aggregating (P < 0.001) and normal-aggregating (P < 0.05) conditions. Although the NO scavenging by RBCs could be impaired with a thicker CFL at higher levels of aggregation, the overall decrease in NO production due to reduction of wall shear stress with the thicker CFL dominantly limited the NO availability in tissue. On the other hand, the O2 availability in tissue increased due to the relatively high core hematocrit in the blood lumen with the thicker CFL.

Effect of Erythrocyte Aggregation on Spatiotemporal Variations in Cell‐Free Layer Formation Near on Arteriolar Bifurcation

To investigate how red blood cell aggregation could modulate the spatial variations in cell-free layer formation in the vicinity of an arteriolar bifurcation. Visualization of blood flow was performed in upstream and downstream vessels of arteriolar bifurcations in the rat cremaster muscles under reduced flow conditions before and after induction of red blood cell aggregation to both physiological normal- and pathological hyperlevels seen in humans. Large asymmetries of layer widths on opposite sides of the downstream vessel were attenuated along the vessel and this effect could be prominently enhanced by the hyperaggregation due to a higher formation rate of the layer which was greater on one side than the other of the vessel. The proportion of downstream layer formation constituted by the smaller downstream vessel generally increased with a thicker layer width at the wall of the upstream vessel adjacent it. A greater tendency of the layer formation in the smaller downstream vessel was found under the hyperaggregating condition than normal-aggregating and nonaggregating conditions. Red blood cell aggregation could attenuate the asymmetry in cell-free layer formation on opposite sides of the downstream vessel, but enhances the heterogeneity of the layer formation between downstream vessels.

Temporal variations of the cell-free layer width may enhance NO bioavailability in small arterioles: Effects of erythrocyte aggregation

Recently, we have shown that temporal variations in the cell-free layer width can potentially enhance nitric oxide (NO) bioavailability in small arterioles. Since the layer width variations can be augmented by red blood cell aggregation, we tested the hypothesis that an increase in the layer width variations due to red blood cell aggregation could provide an underlying mechanism to improve NO bioavailability in the endothelium and promote vasodilatory effects. Utilizing cell-free layer width data acquired from arterioles of the rat cremaster muscle before and after dextran infusion in reduced flow conditions (wall shear stress = 0.13–0.24 Pa), our computational model predicted exponential enhancements of NO bioavailability in the endothelium and soluble guanylyl cyclase (sGC) activation in the smooth muscle layer with increasing temporal variability of the layer width. These effects were mediated primarily by the transient responses of wall shear stress and NO production rate to the layer width variations. The temporal variations in the layer width were significantly enhanced ( P < 0.05) by aggregation, leading to significant improvements ( P < 0.05) in NO bioavailability and sGC activation. As a result, the significant reduction ( P < 0.05) of sGC activation due to the increased width of the layer after aggregation induction was diminished by the opposing effect of the layer variations. These findings highlighted the possible enhancement of NO bioavailability and vascular tone in the arteriole by the augmented layer width variations due to the aggregation.

Effect of erythrocyte aggregation and flow rate on cell-free layer formation in arterioles

Formation of a cell-free layer is an important dynamic feature of microcirculatory blood flow, which can be influenced by rheological parameters, such as red blood cell aggregation and flow rate. In this study, we investigate the effect of these two rheological parameters on cell-free layer characteristics in the arterioles (20-60 mum inner diameter). For the first time, we provide here the detailed temporal information of the arteriolar cell-free layer in various rheological conditions to better describe the characteristics of the layer variation. The rat cremaster muscle was used to visualize arteriolar flows, and the extent of aggregation was raised by dextran 500 infusion to levels seen in normal human blood. Our results show that cell-free layer formation in the arterioles is enhanced by a combination of flow reduction and red blood cell aggregation. A positive relation (P < 0.005) was found between mean cell-free layer widths and their corresponding SDs for all conditions. An analysis of the frequency and magnitudes of cell-free layer variation from their mean value revealed that the layer deviated with significantly larger magnitudes into the red blood cell core after flow reduction and dextran infusion (P < 0.05). In accordance, the disparity of cell-free layer width distribution found in opposite radial directions from its mean became greater with aggregation in reduced flow conditions. This study shows that the cell-free layer width in arterioles is dependent on both flow rate and red blood cell aggregability, and that the temporal variations in width are asymmetric with a greater excursion into the red blood cell core than toward the vessel wall.

Effect of erythrocyte aggregation and flow rate on temporal variation of cell‐free layer width in arterioles

The cell‐free layer (CFL) is a dynamic feature of microcirculatory blood flow and its variability is commonly described in terms of standard deviation (SD). However, CFL width in an arteriole follows a non‐Gaussian distribution imposed by asymmetrical nature of its physical environment, which implies that more detailed statistical analyses are required to better interpret CFL characteristics. In this study, rat cremaster muscle was used to visualize arteriolar flows and erythrocyte aggregability was raised by dextran infusion to levels seen in normal human blood. The effects of aggregation and flow reduction on CFL characteristics in arterioles (ID 20‐50 μm) were investigated. A positive relation was found between mean CFL widths and their corresponding SD and was predominantly enhanced by flow reduction. Both normalized mean and SD of CFL width increased significantly at low pseudoshear rates (17.1 ± 6.1 s−1), especially in the presence of aggregation. The variability of the CFL width was further analyzed in terms of its frequency and magnitude of deviations from its mean value, extending either towards the vessel wall or into the erythrocyte core. There was no significant effect of either flow condition or aggregation on the frequency of CFL deviations, whereas the magnitude of deviations significantly increased after flow reduction and dextran infusion (P &lt; 0.05), especially into the erythrocyte core.

Effects of erythrocyte aggregation and venous network geometry on red blood cell axial migration

Axial migration of red blood cells in small glass tubes can cause blood viscosity to be effectively independent of shear rate. However, this phase separation may not occur to the same degree in the venous network due to infusion of cells and aggregates at branch points. To investigate this hypothesis, we followed trajectories of fluorescently labeled red blood cells in the venular network of the rat spinotrapezius muscle at normal and reduced flow with and without red blood cell aggregation. Cells traveling near the wall of an unbranched venular segment migrated approximately 1% of the longitudinal path length without aggregation and migrated slightly more with aggregation. Venular segment length between branch points averaged three to five times the diameter. Cells in the main vessel were shifted centrally by up to 20% of diameter at branch points, reducing the migration rate of cells near the opposite wall to <1% even in the presence of aggregation. We conclude that formation of a cell-free marginal layer in the venular network is attenuated due to the time dependence of axial migration and the frequent branching of the network.

O(2) release from erythrocytes flowing in a narrow O(2)-permeable tube: effects of erythrocyte aggregation.

The effects of erythrocyte aggregation on O(2) release were examined using O(2)-permeable fluorinated ethylenepropylene copolymer tubes (inner diameter, 25 microm; outer diameter, 100 microm). Measurements were performed using an apparatus built on an inverted microscope that contained a scanning-grating spectrophotometer with a photon count detector connected to two photomultipliers and an image processor through a video camera. The rate of O(2) release from the cells flowing in the narrow tube was determined based on the visible absorption spectrum and the flow velocity of the cells as well as the tube size. When the tube was exposed to nitrogen-saturated deoxygenated saline containing 10 mM sodium dithionite, the flowing erythrocytes were deoxygenated in proportion to the traveling distance, and the deoxygenation at a given distance increased with decreasing flow velocity and cell concentration (hematocrit). Adding Dextran T-70 to the cell suspension increased erythrocyte aggregation in the tube, which resulted in suppressed cell deoxygenation and increased marginal cell-free-layer thickness. The deoxygenation was inversely proportional to the cell-free-layer thickness. The relation was not essentially altered even when the medium viscosity was adjusted with Dextran T-40 to remain constant. The rate of O(2) release from erythrocytes in the tube was discussed in relation to the O(2) diffusion process. We conclude that the diffusion of O(2) from erythrocytes flowing in narrow tubes is inhibited primarily by erythrocyte aggregation itself and partly by thickening of the cell-free layer.

Effect of erythrocyte aggregation on velocity profiles in venules.

A recent whole organ study in cat skeletal muscle showed that the increase in venous resistance seen at reduced arterial pressures is nearly abolished when the muscle is perfused with a nonaggregating red blood cell suspension. To explore a possible underlying mechanism, we tested the hypothesis that red blood cell aggregation alters flow patterns in vivo and leads to blunted red blood cell velocity profiles at reduced shear rates. With the use of fluorescently labeled red blood cells in tracer quantities and a video system equipped with a gated image intensifier, we obtained velocity profiles in venous microvessels (45-75 microm) of rat spinotrapezius muscle at centerline velocities between 0.3 and 14 mm/s (pseudoshear rates 3-120 s(-1)) under normal (nonaggregating) conditions and after induction of red blood cell aggregation with Dextran 500. Profiles are nearly parabolic (Poiseuille flow) over this flow rate range in the absence of aggregation. When aggregation is present, profiles are parabolic at high shear rates and become significantly blunted at pseudoshear rates of 40 s(-1) and below. These results indicate a possible mechanism for increased venous resistance at reduced flows.

Quantitative evaluation of flow dynamics of erythrocytes in microvessels: influence of erythrocyte aggregation.

Effects of erythrocyte aggregation on the flow dynamics of erythrocytes in microvessels were examined quantitatively by perfusing human erythrocytes suspended in isotonic medium containing various concentrations of dextran (70,400 avg mol wt, Dx-70) into a part of the microvascular bed isolated from rabbit mesentery. Thickness of the marginal cell-free layer was measured with an image analyzer, total flow resistance was determined on the basis of the perfusion pressure-volume flow relationship, and homogeneity of erythrocyte flow was evaluated by the power spectrum obtained by the fast Fourier transform of the light intensity change monitored on single microvessels. With increasing dextran concentration, suspension viscosity of erythrocytes at high shear rates increased linearly and thickness of the cell-free layer increased in a sigmoidal fashion. Flow resistance increased relatively little over the range of dextran concentrations in which the cell-free layer increased most rapidly. Furthermore, the flow pattern of erythrocytes in microvessels became inhomogeneous. In conclusion, the present study shows that Dx-70-induced erythrocyte aggregation results in increased flow resistance in the circulatory system, even through the widening of the cell-free layer tends to reduce the resistance and also results in inhomogeneous flow of erythrocytes in microvessels.

The Effect of Erythrocyte Aggregation on the Rheology of Blood

The Effect of Erythrocyte Aggregation on the Rheology of Blood