Efstathios Kaliviotis

The effect of red blood cell aggregation on velocity and cell-depleted layer characteristics of blood in a bifurcating microchannel

Red blood cell (RBC) aggregation is a multifaceted phenomenon, and whether it is generally beneficial or deleterious remains unclear. In order to better understand its effect on microvascular blood flow, the phenomenon must be studied in complex geometries, as it is strongly dependent on time, flow, and geometry. The cell-depleted layer (CDL) which forms at the walls of microvessels has been observed to be enhanced by aggregation; however, details of the characteristics of the CDL in complex regions, such as bifurcations, require further investigation. In the present study, a microchannel with a T-junction was used to analyze the influence of aggregation on the flow field and the CDL. Micro-PIV using RBCs as tracers provided high resolution cell velocity data. CDL characteristics were measured from the same data using a newly developed technique based on motion detection. Skewed and sharpened velocity profiles in the daughter branches were observed, contrary to the behavior of a continuous Newtonian fluid. RBC aggregation was observed to increase the skewness, but decrease the sharpening, of the velocity profiles in the daughter branches. The CDL width was found to be significantly greater, with a wider distribution, in the presence of aggregation and the mean width increased proportionally with the reciprocal of the fraction of flow entering the daughter branch. Aggregation also significantly increased the roughness of the interface between the CDL and the RBC core. The present results provide further insight into how RBC aggregation may affect the flow in complex geometries, which is of importance in both understanding its functions invivo, and utilizing it as a tool in microfluidic devices.

Large scale simulation of red blood cell aggregation in shear flows

Aggregation of highly deformable red blood cells (RBCs) significantly affects the blood flow in the human circulatory system. To investigate the effect of deformation and aggregation of RBCs in blood flow, a mathematical model has been established by coupling the interaction between the fluid and the deformable solids. The model includes a three-dimensional finite volume method solver for incompressible viscous flows, the combined finite-discrete element method for computing the deformation of the RBCs, a JKR model-Johnson, Kendall and Roberts (1964-1971) (Johnson et al., 1971) to take account of the adhesion forces between different RBCs and an iterative direct-forcing immersed boundary method to couple the fluid-solid interactions. The flow of 49,512 RBCs at 45% concentration under the influence of aggregating forces was examined, improving the existing knowledge on simulating flow and structural characteristics of blood at a large scale: previous studies on the particular issue were restricted to simulating the flow of 13,000 aggregative ellipsoidal particles at a 10% concentration. The results are in excellent agreement with experimental studies. More specifically, both the experimental and the simulation results show uniform RBC distributions under high shear rates (60-100/s) whereas large aggregation structures were observed under a lower shear rate of 10/s. The statistical analysis of the simulation data also shows that the shear rate has significant influence on both the flow velocity profiles and the frequency distribution of the RBC orientation angles.

Hematocrit, viscosity and velocity distributions of aggregating and non-aggregating blood in a bifurcating microchannel

Microscale blood flow is characterised by heterogeneous distributions of hematocrit, viscosity and velocity. In microvascular bifurcations, cells are unevenly distributed between the branches, and this effect can be amplified in subsequent branches depending on a number of parameters. We propose an approach to infer hematocrit profiles of human blood flowing through a bifurcating microchannel. The influence of aggregation, induced by the addition of Dextran 2000 to the samples, is also considered. Averaged values indicate plasma skimming, particularly in the presence of red blood cell (RBC) aggregation. Using an empirical model, the hematocrit profiles are used to estimate local relative viscosity distributions. Simulations are used to predict how the non-uniform viscosity influences the velocity profiles. Comparing these data to velocity profiles of RBCs measured using particle image velocimetry provides validation of the model. It is observed that aggregation blunts velocity profiles after a long straight section of channel. Downstream of the bifurcation, skewing of the velocity profiles is detected, which is enhanced by aggregation. The proposed methodology is capable of providing hitherto unreported information on important aspects of microscale blood rheology.

Fast response characteristics of red blood cell aggregation

The present work reports on an important feature of the fast response dynamics of blood flow observed after abrupt changes of the shearing conditions: distinctive peak values in conductance and light reflection/transmission have been observed at short times after the abrupt changes in the shearing conditions and have been attributed to red blood cell (RBC) disorientation and shape changes. Optical shearing microscopy results from the present study show that this peak is directly related to the inter-cellular or inter-aggregate spacing, quantified as the plasma gaps present in the captured images. In order to provide a more in-depth understanding of the structural characteristics of blood subjected to abrupt changes in the flow conditions, normal human blood samples at hematocrits of 45, 35, 25 and 10% were sheared at 100 s(-1) and the shear then suddenly reduced to values decreasing from 60 to 0 s(-1). Results from the present study agree qualitatively and quantitatively with results previously reported in the literature: the hematocrit and the magnitude of the final shear rate affect the magnitude of the peak values. The characteristic peak time was mostly influenced by the cell concentration. It is suggested that aggregation forces may play a part in the process of the fast response structural and spatial rearrangements of RBC.

Spatial variation of blood viscosity: Modelling using shear fields measured by a μPIV based technique

The spatial characteristics of blood viscosity were investigated by combining a newly developed constitutive equation with shear deformation fields calculated from velocity measurements obtained by a μPIV based technique. Blood at physiological hematocrit levels and in the presence of aggregation was sheared in a narrow gap plate-plate geometry and the velocity and aggregation characteristics were determined from images captured using a high resolution camera. Changes in the microstructure of blood caused by aggregation were observed to affect the flow characteristics. At low shear rates, high aggregation and network formation caused the RBC motion to become essentially two-dimensional. The measured velocity fields were used to estimate the magnitude of shear which was subsequently used in conjunction with the new model to assess the spatial variation of viscosity across the flow domain. It was found that the non-uniform microstructural characteristics of blood influence its viscosity distribution accordingly. The viscosity of blood estimated in the core of the examined flow, using a zero-gradient core velocity profile assumption, was found to be significantly higher than the overall effective viscosity determined using other velocity profile assumptions.

Coupled human erythrocyte velocity field and aggregation measurements at physiological haematocrit levels

Simultaneous measurement of erythrocyte (RBC) velocity fields and aggregation properties has been successfully performed using an optical shearing microscope and Particle Image Velocimetry (PIV). Blood at 45% haematocrit was sheared at rates of 5.4< or =gamma < or = 252 s(-1) and imaged using a high speed camera. The images were then processed to yield aggregation indices and flow velocities. Negligible levels of aggregation were observed for gamma > or = 54.0 s(-1), while high levels of aggregation and network formation occurred for gamma < or = 11.7 s(-1). The results illustrate that the velocity measurements are dependent on the extent of RBC aggregation. High levels of network formation cause the velocities at gamma > or = 5.4 s(-1) to deviate markedly from the expected solid body rotation profile. The effect of aggregation level on the PIV accuracy was assessed by monitoring the two-dimensional (2D) correlation coefficients. Lower levels of aggregation result in poorer image correlation, from which it can be inferred that PIV accuracy is reduced. Moreover, aggregation is time-dependent, and consequently PIV accuracy may decrease during recording as the cells break up. It is therefore recommended that aggregation and its effects are taken into account in future when undertaking blood flow studies using PIV. The simplicity of the technique, which requires no lasers, filters, or special pretreatments, demonstrates the potential wide-spread applicability of the data acquisition system for accurate blood flow PIV and aggregation measurement.

Spatial Distributions of Red Blood Cells Significantly Alter Local Haemodynamics

Although bulk changes in red blood cell concentration between vessels have been well characterised, local distributions are generally overlooked. Red blood cells aggregate, deform and migrate within vessels, forming heterogeneous distributions which have considerable effect on local haemodynamics. The present study reports data on the local distribution of human red blood cells in a sequentially bifurcating microchannel, representing the branching geometry of the microvasculature. Imaging methodologies with simple extrapolations are used to infer three dimensional, time-averaged velocity and haematocrit distributions under a range of flow conditions. Strong correlation between the bluntness of the velocity and haematocrit profiles in the parent branch of the geometry is observed and red blood cell aggregation has a notable effect on the observed trends. The two branches of the first bifurcation show similar characteristics in terms of the shapes of the profiles and the extent of plasma skimming, despite the difference in geometric configuration. In the second bifurcation, considerable asymmetry between the branches in the plasma skimming relationship is observed, and elucidated by considering individual haematocrit profiles. The results of the study highlight the importance of considering local haematocrit distributions in the analysis of blood flow and could lead to more accurate computational models of blood flow in microvascular networks. The experimental approaches developed in this work provide a foundation for further examining the characteristics of microhaemodynamics.

Erythrocyte aggregation at non-steady flow conditions: A comparison of characteristics measured with electrorheology and image analysis

In the present study electro-rheology (Contraves LS30 viscometer-based system) and optical shearing microscopy (Lincam CSS450 system and image analysis) techniques have been utilized in order to provide quantitative data on the behaviour of the microstructural properties of whole normal human blood at non-steady flow conditions. The objective of this work is to contribute towards a better understanding of red blood cell aggregation at flow conditions similar to that occurring in a circulatory system and to aid the interpretation and validation of electro-rheological data through a quantitative comparison with data acquired with optical shearing microscopy. Electro-rheology is a promising technique that has been used to provide bulk fluid properties, showing potential for basic research and diagnostic purposes, whereas optical shearing techniques offer a direct assessment of blood microstructure at a cellular level. However, little information exists in the literature regarding the relationships between electro-rheological measurements and blood microstructural characteristics. The results showed that the different non-steady flow conditions affect differently the dynamics of aggregation varying from a parabolic-decrease to an inverted S-shape curve with time. For a wide range of the non-steady flows results obtained with the two different techniques agree to a difference between 1.2 and 12%.

On the effect of microstructural changes of blood on energy dissipation in Couette flow.

Red blood cell aggregation affects the flow of blood at low shear rates; not only the behaviour of the fluid deviates from its Newtonian characteristics, but, depending on the shearing history of the flow, the non-Newtonian characteristics may be influenced. It is not clear how the time and flow-dependent characteristics of the microstructural network developed in blood affect its mechanical properties. The present study aims to improve understanding of the effect of dynamic flow conditions on microstructural characteristics and consequently on the mechanical properties of the fluid. Viscosity measurements on blood samples from healthy volunteers (H=0.45) were taken with a double-walled Couette rheometric cell, under unsteady and quasi-unsteady flow conditions. The aggregation extent index A(alpha), and the microstructural integrity index A(I) were assessed with an optical shearing system and image analysis. Results showed that energy losses in Couette geometries may depend on the structural integrity of the developed RBC network.

Partitioning of red blood cell aggregates in bifurcating microscale flows

Microvascular flows are often considered to be free of red blood cell aggregates, however, recent studies have demonstrated that aggregates are present throughout the microvasculature, affecting cell distribution and blood perfusion. This work reports on the spatial distribution of red blood cell aggregates in a T-shaped bifurcation on the scale of a large microvessel. Non-aggregating and aggregating human red blood cell suspensions were studied for a range of flow splits in the daughter branches of the bifurcation. Aggregate sizes were determined using image processing. The mean aggregate size was marginally increased in the daughter branches for a range of flow rates, mainly due to the lower shear conditions and the close cell and aggregate proximity therein. A counterintuitive decrease in the mean aggregate size was apparent in the lower flow rate branches. This was attributed to the existence of regions depleted by aggregates of certain sizes in the parent branch, and to the change in the exact flow split location in the T-junction with flow ratio. The findings of the present investigation may have significant implications for microvascular flows and may help explain why the effects of physiological RBC aggregation are not deleterious in terms of in vivo vascular resistance.