Red Blood Cell Model Research Articles

Erythrophagocytosis-induced ferroptosis contributes to pulmonary microvascular thrombosis and thrombotic vascular remodeling in pulmonary arterial hypertension

BackgroundWhether primary or just as a complication from the progression of pulmonary arterial hypertension (PAH), thrombosis seems to be an important player in this condition. The crosstalk between red blood cells (RBCs) and pulmonary microvascular endothelial cells (PMVECs) and their role in PAH remain undefined. ObjectivesThe goals of this study were to assess the role of RBC–PMVEC interaction in microvascular thrombosis and thrombotic vascular remodeling under hypoxic conditions. MethodsWe established an in vitro hypoxic coincubation model of RBC and PMVEC as well as a hypoxic mouse model. We investigated erythrophagocytosis (EP), ferroptosis, thrombosis tendency, and pulmonary hemodynamics in experimental PAH. ResultsIncreased EP in PMVEC triggered ferroptosis, enhanced procoagulant activity, and exacerbated vessel remodeling under hypoxic conditions. In the PAH mouse model induced by chronic hypoxia, EP-induced ferroptosis followed by upregulated TMEM16F led to a high tendency of thrombus formation and thrombotic vascular remodeling. Inhibition of ferroptosis or silencing of TMEM16F could alleviate hypercoagulable phenotype, reverse right ventricular systolic pressure, right ventricular hypertrophy index, and remodeling of pulmonary vessels. ConclusionThese results illustrate the pathogenic RBC–PMVEC interactions in PAH. Inhibition EP, ferroptosis, or TMEM16F could be a novel therapeutic target to prevent PAH development and thrombotic complications.

Ginseng total saponin improves red blood cell oxidative stress injury by regulating tyrosine phosphorylation and glycolysis in red blood cells

BackgroundOxidative stress is the main cause of many diseases, but because of its complex pathogenic factors, there is no clear method for treating it. Ginseng total saponin (GTS) an important active ingredients in Panax ginseng C.A. Mey (PG) and has potential therapeutic ability for oxidative stress due to various causes. However, the molecular mechanism of GTS in the treating oxidative stress damage in red blood cells (RBCs) is still unclear. PurposeThis study aimed to examine the protective effect of GTS on RBCs under oxidative stress damage and to determine its potential mechanism. MethodsThe oxidative stress models of rat RBCs induced by hydrogen peroxide (H2O2) and exhaustive swimming in vivo and in vitro was used. We determined the cell morphology, oxygen carrying capacity, apoptosis, antioxidant capacity, and energy metabolism of RBCs. The effect of tyrosine phosphorylation (pTyr) of Band 3 protein on RBCs glycolysis was also examined. ResultsGTS reduced the hemolysis of RBCs induced by H2O2 at the lowest concentration. Moreover, GTS effectively improved the morphology, enhanced the oxygen carrying capacity, and increased antioxidant enzyme activity, adenosine triphosphate (ATP) levels, and adenosine triphosphatase (ATPase) activity in RBCs. GTS also promoted the expression of membrane proteins in RBCs, inhibited pTyr of Band 3 protein, and further improved glycolysis, restoring the morphological structure and physiological function of RBCs. ConclusionsThis study shows, that GTS can protect RBCs from oxidative stress damage by improving RBCs morphology and physiological function. Changes in pTyr expression and its related pTyr regulatory enzymes before and after GTS treatment suggest that Band 3 protein is the main target of GTS in the treating endogenous and exogenous oxidative stress. Moreover, GTS can enhance the glycolytic ability of RBCs by inhibiting pTyr of Band 3 protein, thereby restoring the function of RBCs.

Energy spectrum analysis on a red blood cell model.

It is important to understand the dynamics of red blood cells (RBCs) in blood flow. This requires the formulation of coarse-grained RBC models that reproduce the hydrodynamic properties of blood accurately. One of the models that successfully reproduces the rheology and morphology of blood has been proposed by Fedosov et al. [Comput. Methods Appl. Mech. Eng. 199, 1937-1948 (2010)]. The proposed RBC model contains several parameters whose values are determined by either various experiments or physical requirements. In this study, we developed a new method of determining parameter values precisely from the fluctuations of the RBC membrane. Specifically, we studied the relationship between the spectra of the fluctuations and model parameters. Characteristic peaks were observed in the spectra, whose peak frequencies were dependent on the parameter values. In addition, we investigated the spectra of the radius of gyration. We identified the peaks originating from the spring potential and the volume-conserving potential appearing in the spectra. These results lead to the precise experimental determination of the parameters used in the RBC model.

High-precision non-invasive RBC and HGB detection system based on spectral analysis.

Non-invasive blood composition analysis based on dynamic spectrum (DS) theory has gained significant attention due to its non-invasive, simple, and fast performance. However, most of the multi-wavelength photoplethysmography (PPG) detection devices used to obtain DS are composed of halogen light sources and spectrometers and cannot detect effective PPG signals in the visible light short band (400-620nm), which limits the detection accuracy of blood components with significant absorption spectral differences in that band. Therefore, this paper designs a multi-wavelength spectral acquisition system that can measure high signal-to-noise ratio (SNR > 65dB) PGG signals at wavelengths of 405, 430, 450, 505, 520, and 570nm and combines this system with a halogen lamp spectrometer acquisition system for non-invasive blood component detection. Furthermore, this paper collects the DS of 272 subjects with the combined system and establishes a predictive model for DS with the content of red blood cell (RBC) and hemoglobin (HGB) components. The results show that, compared with the halogen lamp spectrometer acquisition system, the correlation coefficient (Rp) of RBC and HGB prediction model established by the combined system has increased by 0.0619 and 0.0489, respectively, and the root mean square error (RMSE) has decreased by 0.08 1e12/L and 0.85g/L, which confirm the feasibility of the designed multi-wavelength spectrum acquisition system to enhance the accuracy of blood component detection.

Ferroptosis of Endothelial Cells Triggered by Erythrophagocytosis Contributes to Thrombogenesis in Uremia.

Although thrombosis events are the leading complication of uremia, their mechanism is largely unknown. The interaction between endothelial cells (ECs) and red blood cells (RBCs) in uremic solutes and its prothrombotic role need to be investigated. Here, we established an in vitro co-incubation model of uremic RBC and EC as well as a uremic rat model induced by adenine. Using flow cytometry, confocal microscopy, and electron microscopy, we found increased erythrophagocytosis by EC accompanied by increased reactive oxygen species, lipid peroxidation, and impairment of mitochondria, indicating that ECs undergo ferroptosis. Further investigations showed increased proteins' expression of heme oxygenase-1 and ferritin and labile iron pool accumulation in EC, which could be suppressed by deferoxamine (DFO). The ferroptosis-negative regulators glutathione peroxidase 4 and SLC7A11 were decreased in our erythrophagocytosis model and could be enhanced by ferrostatin-1 or DFO. In vivo, we observed that vascular EC phagocytosed RBC and underwent ferroptosis in the kidney of the uremic rat, which could be inhibited by blocking the phagocytic pathway or inhibiting ferroptosis. Next, we found that the high tendency of thrombus formation was accompanied by erythrophagocytosis-induced ferroptosis in vitro and in vivo. Importantly, we further revealed that upregulated TMEM16F expression mediated phosphatidylserine externalization on ferroptotic EC, which contributed to a uremia-associated hypercoagulable state. Our results indicate that erythrophagocytosis-triggered ferroptosis followed by phosphatidylserine exposure of EC may play a key role in uremic thrombotic complications, which may be a promising target to prevent thrombogenesis of uremia.

The stress-free state of human erythrocytes: Data-driven inference of a transferable RBC model

The stress-free state (SFS) of red blood cells (RBCs) is a fundamental reference configuration for the calibration of computational models, yet it remains unknown. Current experimental methods cannot measure the SFS of cells without affecting their mechanical properties, whereas computational postulates are the subject of controversial discussions. Here, we introduce data-driven estimates of the SFS shape and the visco-elastic properties of RBCs. We employ data from single-cell experiments that include measurements of the equilibrium shape of stretched cells and relaxation times of initially stretched RBCs. A hierarchical Bayesian model accounts for these experimental and data heterogeneities. We quantify, for the first time, the SFS of RBCs and use it to introduce a transferable RBC (t-RBC) model. The effectiveness of the proposed model is shown on predictions of unseen experimental conditions during the inference, including the critical stress of transitions between tumbling and tank-treading cells in shear flow. Our findings demonstrate that the proposed t-RBC model provides predictions of blood flows with unprecedented accuracy and quantified uncertainties.

Prior immunization against an intracellular antigen enhances subsequent red blood cell alloimmunization in mice

AbstractAntibodies against red blood cell (RBC) alloantigens can increase morbidity and mortality among transfusion recipients. However, alloimmunization rates can vary dramatically, as some patients never generate alloantibodies after transfusion, whereas others not only become alloimmunized but may also be prone to generating additional alloantibodies after subsequent transfusion. Previous studies suggested that CD4 T–cell responses that drive alloantibody formation recognize the same alloantigen engaged by B cells. However, because RBCs express numerous antigens, both internally and externally, it is possible that CD4 T–cell responses directed against intracellular antigens may facilitate subsequent alloimmunization against a surface RBC antigen. Here, we show that B cells can acquire intracellular antigens from RBCs. Using a mouse model of donor RBCs expressing 2 distinct alloantigens, we demonstrate that immune priming to an intracellular antigen, which would not be detected by any currently used RBC compatibility assays, can directly influence alloantibody formation after exposure to a subsequent distinct surface RBC alloantigen. These findings suggest a previously underappreciated mechanism whereby transfusion recipient responders may exhibit an increased rate of alloimmunization because of prior immune priming toward intracellular antigens.

Lingering Dynamics of Type 2 Diabetes Mellitus Red Blood Cells in Retinal Arteriolar Bifurcations.

It has been proven that the deformability of red blood cells (RBC) is reduced owing to changes in mechanical properties, such as diabetes mellitus and hypertension. To probe the effects of RBC morphological and physical parameters on the flow field in bifurcated arterioles, three types of RBC models with various degrees of biconcave shapes were built based on the in vitro experimental data. The dynamic behaviors of the RBCs in shear flow were simulated to validate the feasibility of the finite element-Arbitrary Lagrangian-Eulerian method with a moving mesh. The influences of the shear rate and viscosity ratios on RBC motions were investigated. The motion of RBCs in arteriolar bifurcations was further simulated. Abnormal variations in the morphological and physical parameters of RBCs may lead to diminished tank-tread motion and enhanced tumbling motion in shear flow. Moreover, abnormal RBC variations can result in slower RBC motion at the bifurcation with a longer transmit time and greater flow resistance, which may further cause inadequate local oxygen supply. These findings would provide useful insights into the microvascular complications in diabetes mellitus.

On‐chip magnetophoretic capture in a model of malaria‐infected red blood cells

The search for new rapid diagnostic tests for malaria is a priority for developing an efficient strategy to fight this endemic disease, which affects more than 3 billion people worldwide. In this study, we characterize systematically an easy‐to‐operate lab‐on‐chip, designed for the magnetophoretic capture of malaria‐infected red blood cells (RBCs). The method relies on the positive magnetic susceptibility of infected RBCs with respect to blood plasma. A matrix of nickel posts fabricated in a silicon chip placed face down is aimed at attracting infected cells, while healthy cells sediment on a glass slide under the action of gravity. Using a model of infected RBCs, that is, erythrocytes with methemoglobin, we obtained a capture efficiency of about 70% after 10 min in static conditions. By proper agitation, the capture efficiency reached 85% after just 5 min. Sample preparation requires only a 1:10 volume dilution of whole blood, previously treated with heparin, in a phosphate‐buffered solution. Nonspecific attraction of untreated RBCs was not observed in the same time interval.

Multiscale computational framework for predicting viscoelasticity of red blood cells in aging and mechanical fatigue

Red blood cells (RBCs) experience significant cyclic deformation through large elastic stretching and relaxation as they circulate in the bloodstream. Such hundreds of thousands times cell deformation causes cumulative fatigue damage to the circulating RBCs, leading to alterations in cell deformability and membrane viscoelasticity. Here, we employ a two-step multiscale computational framework based on coarse-grained molecular dynamics (CGMD) and dissipative particle dynamics (DPD) to probe the viscoelastic properties of the surface-altered RBCs in aging and mechanical fatigue, using experimental information on the molecular structures of both RBC membrane and cytosolic Hb as well as Hb concentration of RBC cytosol. We perform CGMD simulations to compute the shear modulus and membrane viscosity of a small RBC patch with altered horizontal connectivity within spectrin network; meanwhile, we compute the shear viscosity of RBC cytosol under different Hb concentrations. We then use these computed parameters as inputs for a more coarse-grained DPD-based RBC model at the whole-cell level to probe cell deformation, dynamic relaxation, viscoelastic performance of the altered RBC membrane. We find that the RBC membrane rigidity increases with the enhanced horizontal connectivity within spectrin network, but by a factor less than the effective membrane viscosity, resulting in an elevated characteristic relaxation time with increasing fatigue cycles. In addition, we quantify the two-dimensional storage and loss moduli of RBC membrane under time-dependent loading, revealing that the loss modulus is much larger than the storage modulus throughout the entire frequency range covered by the simulations, especially under enhanced connectivity in the cytoskeletal network, which is analogous to the performance of typical viscoelastic materials. These quantitative findings provide unique insights into the progressive damage of circulating RBCs, and demonstrate the capability of the two-step multiscale framework in effectively predicting the altered viscoelastic properties of RBCs influenced by in vivo aging and mechanical fatigue.

Моделювання осциляції поверхонь клітин крові як заповнених рідиною багатошарових в’язкопружних оболонок

Rheological properties of the red blood cells (RBC) determine their movement in the larger and smaller blood vessels, oxygen and carbon dioxide delivery to/from the cells. Those properties vary significantly with age and health state of an organism. In this paper a new rheological model of RBC as a thin multilayer shell, which includes the cytoskeleton, lipid bilayer, glycocalyx, and hydrate shell as Maxwell's viscoelastic bodies is proposed. Mechanical properties of the rheological model in isotonic, isometric and dynamic experiments are studied. The oscillations of the surfaces of erythrocytes or other cells in the approximation of multilayer viscoelastic shell filled with a viscous fluid are investigated. The expressions for the dynamic Young’s modules and viscosity/fluidity coefficients as functions of the viscoelastic and geometric parameters of the layers are obtained. The problem of propagation of small perturbations along the cell surface is considered. The solutions of the problem in the form of Young and Lamé waves are obtained. The method of identification of the erythrocyte parameters from the experimental measurements of the wave propagation on the basis of the developed mathematical model for the purposes of clinical diagnostics of diseases with use of a microdrop of blood of the patient is proposed.

Resolved CFD-DEM simulation of blood flow with a reduced-order RBC model

Mathematical modeling of the blood flow with a resolved description of biological cells mechanics such as red blood cell (RBC) has been a challenge in the past decades as it involves physical complexities and demands high computational costs. In the present study, we propose an approach for efficient simulation of blood flow with several suspended RBCs. In this approach, we employ our previously proposed reduced-order model for deformable particles (Nair et al. in Comput Part Mech 7:593–601, 2020) to mimic the mechanical behavior of an individual RBC as a cluster of overlapping spheres interconnected by flexible mathematical bonds. This discrete element method-based model is then coupled with a fluid flow solver using the immersed boundary method with continuous forcing in the context of computational fluid dynamics-discrete element method (CFD-DEM) coupling. The present computational method is tested with a couple of validation cases in which the single RBC dynamics, as well as the blood flow with several RBCs, were tested in comparison with existing literature date. First, the RBC deformation index in shear flow at different shear rates is studied with a good accuracy. Then, the blood flow in micro-tubes of different diameters and hematocrits was simulated. The key characteristics of blood flow such as cell-free layer (CFL) thickness, Fahraeus effect and the relative apparent viscosity are used as the validation metrics. The proposed approach can predict the formation of the migration of RBC toward the tube center-line and the CFL thickness in good agreement with previous measurement and simulations. Furthermore, the model is employed to study the CFL enhancement for plasma separation based on channel constriction. The simulation results compute the CFL thickness downstream of the channel constriction in good agreement with the experiments in a wide range of flow rates and constriction lengths. The original contribution of this study lies in proposing an efficient resolved CFD-DEM simulation method for blood flows with many RBCs which can be employed for numerical investigation of bio-microfluidic applications.

Computational modeling of biomechanics and biorheology of heated red blood cells

Because of their compromised deformability, heat denatured erythrocytes have been used as labeled probes to visualize spleen tissue or to assess the ability of the spleen to retain stiff red blood cells (RBCs) for over three decades, e.g., see Looareesuwan et al. N. Engl. J. Med. (1987). Despite their good accessibility, it is still an open question how heated RBCs compare to certain diseased RBCs in terms of their biomechanical and biorheological responses, which may undermine their effective usage and even lead to misleading experimental observations. To help answering this question, we perform a systematic computational study of the hemorheological properties of heated RBCs with several physiologically relevant static and hemodynamic settings, including optical-tweezers test, relaxation of prestretched RBCs, RBC traversal through a capillary-like channel and a spleen-like slit, and a viscometric rheology test. We show that our in silico RBC models agree well with existing experiments. Moreover, under static tests, heated RBCs exhibit deformability deterioration comparable to certain disease-impaired RBCs such as those in malaria. For RBC traversal under confinement (through microchannel or slit), heated RBCs show prolonged transit time or retention depending on the level of confinement and heating procedure, suggesting that carefully heat-treated RBCs may be useful for studying splenic- or vaso-occlusion in vascular pathologies. For the rheology test, we expand the existing bulk viscosity data of heated RBCs to a wider range of shear rates (1–1000 s−1) to represent most pathophysiological conditions in macro- or microcirculation. Although heated RBC suspension shows elevated viscosity comparable to certain diseased RBC suspensions under relatively high shear rates (100–1000 s−1), they underestimate the elevated viscosity (e.g., in sickle cell anemia) at low shear rates (<10 s−1). Our work provides mechanistic rationale for selective usage of heated RBC as a potentially useful model for studying the abnormal traversal dynamics and hemorheology in certain blood disorders.

Numerical simulation of spatiotemporal red blood cell aggregation under sinusoidal pulsatile flow

Previous studies on red blood cell (RBC) aggregation have elucidated the inverse relationship between shear rate and RBC aggregation under Poiseuille flow. However, the local parabolic rouleaux pattern in the arterial flow observed in ultrasonic imaging cannot be explained by shear rate alone. A quantitative approach is required to analyze the spatiotemporal variation in arterial pulsatile flow and the resulting RBC aggregation. In this work, a 2D RBC model was used to simulate RBC motion driven by interactional and hydrodynamic forces based on the depletion theory of the RBC mechanism. We focused on the interaction between the spatial distribution of shear rate and the dynamic motion of RBC aggregation under sinusoidal pulsatile flow. We introduced two components of shear rate, namely, the radial and axial shear rates, to understand the effect of sinusoidal pulsatile flow on RBC aggregation. The simulation results demonstrated that specific ranges of the axial shear rate and its ratio with radial shear rate strongly affected local RBC aggregation and parabolic rouleaux formation. These findings are important, as they indicate that the spatiotemporal variation in shear rate has a crucial role in the aggregate formation and local parabolic rouleaux under pulsatile flow.

Quantitative prediction of flow dynamics and mechanical retention of surface-altered red blood cells through a splenic slit

Normal red blood cells (RBCs) have remarkable properties of deformability, which enable them to squeeze through tiny splenic inter-endothelial slits (IESs) without any damage. Decreased surface-area-to-volume (SA/V) ratio through the loss of membrane surface is a key determinant of splenic entrapment of surface-altered RBCs due to cell aging or disease. Here, we investigate the flow dynamics and mechanical retention of the surface-altered RBCs with different extents of surface area loss, using a multiscale RBC (MS-RBC) model implemented in dissipative particle dynamics (DPD). We show that the DPD-based MS-RBC simulations can accurately reproduce the ex vivo experimentally measured rate of RBC mechanical retention when we take into account the distribution of RBC surface area (i.e., the size difference within the RBC population). We also examine the cumulative effect of the cell surface area loss on the traversal dynamics of the surface-altered RBCs, where we found that the final values of cell surface area (or the SA/V ratio) play a key role in determining the RBC traversal dynamics, regardless of the loss pathway of cell surface area. Taken together, these simulation results have implications for understanding the sensitivity of the splenic IESs to retain and clear the surface-altered RBCs with increased surface area loss, providing an insight into the fundamental flow dynamics and mechanical clearance of the surface-altered RBCs by the human spleen.

Hierarchical Bayesian Uncertainty Quantification for a Model of the Red Blood Cell

Simulations of blood flows in microfluidic devices and physiological systems are gaining importance in complementing experimental and clinical studies. The predictive capabilities of these simulations hinge on the parameters of the red blood cell (RBC) model that are usually calibrated from experimental data. However, these parameter values may vary drastically when calibrated using different experimental quantities or experimental settings. In turn, the results of existing blood flow simulations largely depend on the utilized parameters that have been chosen to validate a particular experiment. We suggest a revision to this type of model calibration to properly integrate experimental data in the computational models and accordingly inform their predictions. In this context, we introduce the calibration of a popular RBC model using data-driven, hierarchical Bayesian inference. We employ data from classical experiments of RBC stretching by optical tweezers and tank treading in shear flows, and distinguish the calibration of the model parameters through single-level and hierarchical Bayesian uncertainty quantification. We find that the optimal model parameters depend not only on the data used for the inference but also on the way the data are used in the inference process. Single-level Bayesian models predict well the data used in their calibration, but are inferior to the hierarchical Bayesian model at predicting previously unseen data. This work demonstrates that the proper integration of experimental data is essential for the development of a robust and transferable RBC model. We believe that the present study can serve as a prototype across scientific fields, in revising the integration of computational models and heterogeneous experimental data.

Lingering Dynamics in Microvascular Blood Flow

The microvascular networks in the body of vertebrates consist of the smallest vessels such as arterioles, capillaries, and venules. The flow of red blood cells (RBCs) through these networks ensures the gas exchange in as well as the transport of nutrients to the tissues. Any alterations in this blood flow may have severe implications on the health state. Because the vessels in these networks obey dimensions similar to the diameter of RBCs, dynamic effects on the cellular scale play a key role. The steady progression in the numerical modeling of RBCs, even in complex networks, has led to novel findings in the field of hemodynamics, especially concerning the impact and the dynamics of lingering events when a cell meets a branch of the network. However, these results are yet to be matched by a detailed analysis of the lingering experiments in vivo. To quantify this lingering effect in in vivo experiments, this study analyzes branching vessels in the microvasculature of Syrian golden hamsters via intravital microscopy and the use of an implanted dorsal skinfold chamber. It also presents a detailed analysis of these lingering effects of cells at the apex of bifurcating vessels, affecting the temporal distribution of plasmatic zones of blood flow in the branches and even causing a partial blockage in severe cases.

Determining a critical threshold for G6PD activity below which red blood cell response to oxidative stress is poor

BackgroundGlucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme disorder in the world. Its main function is to generate NADPH that is required for anti-oxidative pathway in the cells especially in red blood cells (RBC). G6PD deficiency is X-linked and thus subject to random X-chromosome inactivation in women giving them mosaic expression of G6PD activities in their individual cells. This phenomenon makes it difficult for diagnosis with the currently available G6PD qualitative diagnostic tests. With the rolling out of newly marketed anti-malarial drug tafenoquine, which has a long half-life, screening for G6PD deficiency becomes a necessity where those with < 70% G6PD activity cannot receive this drug. Thus, evidence for a quantitative cut-off for G6PD activity is needed to ensure safe drug administration.MethodsRBC models were developed to analyse the effect of oxidant on RBC oxidative markers namely total glutathione (GSH)and malondialdehyde (MDA). G6PD activity was measured using quantitative assay from Trinity Biotech and was correlated with cytofluorometric assay. RBC from two G6PD heterozygous women with different G6PD activities were also analysed for comparison.ResultsThere was a negative correlation between G6PD activity and CuCl concentration and a strong association between G6PD activities and proportion of G6PD normal RBC in CuCl-treated models and in ex vivo RBC. However, in terms of oxidative stress markers analyses, unlike the hypothesis where the lower G6PD activity, the higher MDA and the lower GSH level, the CuCl RBC model showed that in low G6PD activities (10–30%) cells, the MDA level is lower compared to the rest of the models (p < 0.05). The ex vivo models however were in line with the hypothesis, although the result was not significant (p = 0.5). There was a significant difference between RBC with < 60% and those with > 80% G6PD activities in CuCl RBC model, but not in ex vivo RBC (p = 0.5). Genotyping heterozygous subjects showed G6PDViangchan variant with 2.97 U/gHb (33% activity) and 6.58 U/gHb (74% activity).ConclusionsThe GSH analysis has pointed to the 60% G6PD activity cut-off and this data is supportive of the old World Health Organization threshold for intermediate upper limit of 60% G6PD activity. However, there are significant limitations in using MDA assay with CuCl RBC model because the RBC was already stressed due to the copper treatment and thus present a different result when compared to the ex vivo model.

Prediction of mechanical hemolysis in medical devices via a Lagrangian strain-based multiscale model.

This work introduces a new Lagrangian strain-based model to predict the shear-induced hemolysis in biomedical devices. Current computational models for device-induced hemolysis usually utilize empirical fitting of the released free hemoglobin (Hb) in plasma from damaged red blood cells (RBCs). These empirical correlations contain parameters that depend on specific device and operating conditions, thus cannot be used to predict hemolysis in a general device. The proposed algorithm does not have any empirical parameters, thus can presumably be used for hemolysis prediction in various blood-wetting medical devices. In contrast to empirical correlations in which the Hb release is related to the shear stress and exposure time without considering the physical processes, the proposed model links flow-induced deformation of the RBC membrane to membrane permeabilization and Hb release. In this approach, once the steady-state numerical solution of blood flow in the device is obtained under a prescribed operating condition, sample path lines are traced from the inlet of the device to the outlet to calculate the history of the shear stress tensor. In solving the fluid flow, it is assumed that RBCs do not have any influence on the flow pattern. Along each path line, shear stress tensor will be input into a coarse-grained (CG) RBC model to calculate the RBC deformation. Then the correlations obtained from molecular dynamics (MD) simulations are applied to relate the local areal RBC deformation to the perforated area on the RBC membrane. Finally, Hb released out of transient pores is calculated over each path line via a diffusion equation considering the effects of the steric hindrance and increased hydrodynamic drag due to the size of the Hb molecule. The total index of hemolysis (IH) is calculated by integration of released Hb over all the path lines in the computational domain. Hemolysis generated in the Food and Drug Administration (FDA) nozzle and two blood pumps, that is, a CentriMag blood pump (a centrifugal pump) and HeartMate II (an axial pump), for different flow regimes including the laminar and turbulent flows are calculated via the proposed algorithm. In all the simulations, the numerical predicted IH is close to the range of experimental data. The results promisingly indicate that this multiscale approach can be used as a tool for predicting hemolysis and optimizing the hematologic design of other types of blood-wetting devices.

INVERSE UNCERTAINTY QUANTIFICATION OF A CELL MODEL USING A GAUSSIAN PROCESS METAMODEL

In order to accurately describe the mechanics of red blood cells (RBCs) and resulting fluid dynamics, a cell-resolved blood flow fluid solver is required. The parameters of the material model for the RBC membranes are carefully tuned to reproduce the behavior of real cells under various experimental conditions. In this work, uncertainty in the parameters of the material model for RBCs used in a model for RBC suspensions was estimated with Inverse Uncertainty Quantification (IUQ) using Bayesian Annealed Sequential Importance Sampling (BASIS). Due to the relatively high computational cost of the model, a Gaussian Process regression metamodel was trained in order to feasibly draw the large number of samples required to obtain an accurate posterior distribution estimate. Additionally, the identifiability of the model parameters was estimated using Sobol sensitivity indices. The elongation index of simulated RBCs in a perfect sheared environment was the model prediction used to calibrate model parameters. The results show good identifiability of the parameter defining the tensile properties of the cell membrane and viscosity ratio, and poor identifiability of the parameter defining the response of the cell surface while undergoing bending. This suggests that the latter should be identified using a different quantity of interest. Overall, the model outputs with the optimal values of the parameters obtained using the Gaussian Process metamodel match better or close to the measurements than the results with the parameters' values obtained with the original model. Therefore, we can conclude that it is a valid method to decrease the computational cost of IUQ of the model.