ABSTRACT
BACKGROUND: Oxidative stress is increased in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients and leads to the development of graft versus host disease (GVHD). Mesenchymal stromal cells (MSCs) can ameliorate GVHD by regulating the function of T cells. However, whether MSCs can modulate erythrocyte antioxidant metabolism and thus reduce GVHD is not known. METHODS: Forty female BALB/c mice were randomly assigned to four groups: the control, GVHDhigh, hPMSC, and PBS groups. A hypoxanthine/xanthine oxidase system was used to steadily and gradually produce superoxide in an in vitro experiment. A scanning microscope was used to examine the ultrastructure of erythrocytes. Laser diffraction analyses were used to analyze erythrocyte deformability. Western blotting was used to measure the expression of the erythrocyte membrane skeleton proteins Band 3 and ß-Spectrin. Corresponding kits were used to assess the levels of oxidative damage and the activity of antioxidant enzymes. RESULTS: Morphological and deformability defects were significantly increased in erythrocytes from GVHD patients. Band 3 and ß-Spectrin expression was also reduced in GVHD patients and model mice. Furthermore, we observed significantly increased oxidative stress-induce injury and decreased antioxidant capability in erythrocytes from both GVHD patients and model mice. Subsequent research showed that human placenta-derived MSC (hPMSC) therapy decreased the GVHD-induced redox imbalance in erythrocytes. Furthermore, our findings suggested that upregulating glucose metabolism promoted both the de novo synthesis and recycling of GSH, which is the primary mechanism by which hPMSCs mediate the increase in antioxidant capacity in erythrocytes. CONCLUSION: Together, our findings suggest that hPMSCs can increase antioxidant capacity by increasing erythrocyte GSH production and thus ameliorate GVHD.
Subject(s)
Erythrocytes , Glutathione , Graft vs Host Disease , Mesenchymal Stem Cells , Mice, Inbred BALB C , Oxidative Stress , Animals , Female , Erythrocytes/metabolism , Humans , Mesenchymal Stem Cells/metabolism , Glutathione/metabolism , Mice , Placenta/metabolism , Pregnancy , Mesenchymal Stem Cell Transplantation , Hematopoietic Stem Cell Transplantation/adverse effects , Adult , Cells, Cultured , Middle Aged , Erythrocyte Deformability , Disease Models, AnimalABSTRACT
The deformability of red blood cells (RBCs), expressing their ability to change their shape as a function of flow-induced shear stress, allows them to optimize oxygen delivery to the tissues and minimize their resistance to flow, especially in microcirculation. During physiological aging and blood storage, or under external stimulations, RBCs undergo metabolic and structural alterations, one of which is hemoglobin (Hb) redistribution between the cytosol and the membrane. Consequently, part of the Hb may attach to the cell membrane, and although this process is reversible, the increase in membrane-bound Hb (MBHb) can affect the cell's mechanical properties and deformability in particular. In the present study, we examined the correlation between the MBHb levels, determined by mass spectroscopy, and the cell deformability, determined by image analysis. Six hemoglobin subunits were found attached to the RBC membranes. The cell deformability was negatively correlated with the level of four subunits, with a highly significant inter-correlation between them. These data suggest that the decrease in RBC deformability results from Hb redistribution between the cytosol and the cell membrane and the respective Hb interaction with the cell membrane.
Subject(s)
Erythrocyte Deformability , Erythrocyte Membrane , Hemoglobins , Humans , Erythrocyte Membrane/metabolism , Hemoglobins/metabolism , Erythrocytes/metabolism , Protein BindingABSTRACT
BACKGROUND: Blood collection from donors on testosterone therapy (TT) is restricted to red blood cell (RBC) concentrates to avoid patient exposure to supraphysiological testosterone (T). The objective of this study was to identify TT-related changes in RBC characteristics relevant to transfusion effectiveness in patients. STUDY DESIGN: This was a two-part study with cohorts of patients and blood donors on TT. In part 1, we conducted longitudinal evaluation of RBCs collected before and at three time points after initiation of T. RBC assays included storage and oxidative hemolysis, membrane deformability (elongation index), and oximetry. In part 2, we evaluated the fate of transfused RBCs from TT donors in immunodeficient mice and by retrospective analyses of NIH's vein-to-vein databases. RESULTS: TT increased oxidative hemolysis (1.45-fold change) and decreased RBC membrane deformability. Plasma free testosterone was positively correlated with oxidative hemolysis (r = .552) and negatively correlated with the elongation index (r = -.472). Stored and gamma-irradiated RBCs from TT donors had lower posttransfusion recovery in mice compared to controls (41.6 ± 12 vs. 55.3 ± 20.5%). Recipients of RBCs from male donors taking T had 25% lower hemoglobin increments compared to recipients of RBCs from non-TT male donors, and had increased incidence (OR, 1.80) of requiring additional RBC transfusions within 48 h of the index transfusion event. CONCLUSIONS: TT is associated with altered RBC characteristics and transfusion effectiveness. These results suggest that clinical utilization of TT RBCs may be less effective in recipients who benefit from longer RBC survival, such as chronically transfused patients.
Subject(s)
Blood Preservation , Erythrocyte Deformability , Erythrocyte Transfusion , Erythrocytes , Oxidative Stress , Testosterone , Testosterone/blood , Humans , Male , Oxidative Stress/drug effects , Erythrocytes/metabolism , Erythrocytes/drug effects , Erythrocyte Deformability/drug effects , Animals , Mice , Middle Aged , Female , Hemolysis/drug effects , Adult , Retrospective Studies , Blood Donors , Cell Survival/drug effects , AgedABSTRACT
Type 2 diabetes mellitus (T2DM) is a prevalent and debilitating disease with numerous health risks, including cardiovascular diseases, kidney dysfunction, and nerve damage. One important aspect of T2DM is its association with the abnormal morphology of red blood cells (RBCs), which leads to increased blood viscosity and impaired blood flow. Therefore, evaluating the mechanical properties of RBCs is crucial for understanding the role of T2DM in cellular deformability. This provides valuable insights into disease progression and potential diagnostic applications. In this study, we developed an open micro-electro-fluidic (OMEF) biochip technology based on dielectrophoresis (DEP) to assess the deformability of RBCs in T2DM. The biochip facilitates high-throughput single-cell RBC stretching experiments, enabling quantitative measurements of the cell size, strain, stretch factor, and post-stretching relaxation time. Our results confirm the significant impact of T2DM on the deformability of RBCs. Compared to their healthy counterparts, diabetic RBCs exhibit â¼27% increased size and â¼29% reduced stretch factor, suggesting potential biomarkers for monitoring T2DM. The observed dynamic behaviors emphasize the contrast between the mechanical characteristics, where healthy RBCs demonstrate notable elasticity and diabetic RBCs exhibit plastic behavior. These differences highlight the significance of mechanical characteristics in understanding the implications for RBCs in T2DM. With its â¼90% sensitivity and rapid readout (ultimately within a few minutes), the OMEF biochip holds potential as an effective point-of-care diagnostic tool for evaluating the deformability of RBCs in individuals with T2DM and tracking disease progression.
Subject(s)
Diabetes Mellitus, Type 2 , Erythrocyte Deformability , Erythrocytes , Humans , Diabetes Mellitus, Type 2/diagnosis , Erythrocytes/cytology , Erythrocytes/pathology , Lab-On-A-Chip Devices , Electrophoresis/instrumentation , Microfluidic Analytical Techniques/instrumentation , Equipment DesignABSTRACT
The production of HbS - an abnormal hemoglobin (Hb) - in sickle cell disease (SCD) results in poorly deformable red blood cells (RBCs) that are prone to microcapillary occlusion, causing tissue ischemia and organ damage. Novel treatments, including gene therapy, may reduce SCD morbidity, but methods to functionally evaluate RBCs remain limited. Previously, we presented the microfluidic impedance red cell assay (MIRCA) for rapid assessment of RBC deformability, employing electrical impedance-based readout to measure RBC occlusion of progressively narrowing micropillar openings. We describe herein the design, development, validation, and clinical utility of the next-generation MIRCA assay, featuring enhanced portability, rapidity, and usability. It incorporates a miniaturized impedance analyzer and features a simplified wash-free operation that yields an occlusion index (OI) within 15 min as a new metric for RBC occlusion. We show a correlation between OI and percent fetal hemoglobin (%HbF), other laboratory biomarkers of RBC hemolysis, and SCD severity. To demonstrate the assay's versatility, we tested RBC samples from treatment-naïve SCD patients in Uganda that yielded OI levels similar to those from hydroxyurea (HU)-treated patients in the U.S., highlighting the role of %HbF in protecting against microcapillary occlusion independent of other pharmacological effects. The MIRCA assay could also identify a subset of HU-treated patients with high occlusion risks, suggesting that they may require treatment adjustments including a second-line therapy to improve their outcomes. This work demonstrates the potential of the MIRCA assay for accelerated evaluation of RBC health, function, and therapeutic effect in an ex vivo model of the microcapillary networks.
Subject(s)
Anemia, Sickle Cell , Biosensing Techniques , Electric Impedance , Erythrocytes , Humans , Anemia, Sickle Cell/blood , Biosensing Techniques/instrumentation , Equipment Design , Erythrocyte Deformability , Microfluidic Analytical Techniques/instrumentation , Hemolysis , Lab-On-A-Chip DevicesABSTRACT
OBJECTIVE: To study the dynamics of hemorheologic changes and the frequency of early complications of laparoscopic radical hysterectomy in patients with uterine corpus cancer depending on conducting rehabilitation activities in the early postoperative period. MATERIAL AND METHODS: The number of patients with uterine corpus cancer equal 49 (mean age 54.8±2.2 years), divided into 2 comparable groups, was examined: experimental group - 23 patients, who received local magnetotherapy since the first day after surgery for 5-6 days, and control group - 26 patients without physiotherapy. Comparative group included 24 healthy women. The basic rheological parameters, namely blood viscosity at high and low shear rate, hematocrit, erythrocytes' aggregation and deformability, erythrocytes and platelets electrophoretic mobility, were evaluated in all patients initially, on the 1st and 5th days after surgery and in comparison group. RESULTS: There were changes in the rheological properties of the blood before surgery in patients of both groups: increase of blood viscosity, enhancement of aggregation activity of its formed elements, decrease of erythrocytes' deformability properties. The laparoscopic radical hysterectomy was accompanied by the exacerbation of these disorders. The early magnetotherapy in patients reduced hemorheological abnormalities up to the preoperative parameters (p<0.05) for 5 days, as well as reduced the incidence of early postoperative complications by 2.4 times compared to the control group. CONCLUSION: The application of local low-frequency low-intensity magnetotherapy since the first postoperative day allows to reduce the level of postoperative hemorheological abnormalities up to the level of preoperative parameters, as well as the frequency of early postoperative complications.
Subject(s)
Neoplasms , Humans , Female , Middle Aged , Hemorheology , Erythrocyte Deformability , Erythrocyte Aggregation , Postoperative ComplicationsABSTRACT
BACKGROUND: A challenge for coaches and athletes is to find the best combination of exercises during training. Considering its favorable effects, HIIT has been very popular recently. OBJECTIVE: The goal of this study was to investigate anthropometric features, performance, erythrocyte deformability, plasma viscosity (PV) and oxidative stress in response to acute and long-term (6 weeks) HIIT in adolescent basketball players. METHODS: 22 sportsmen between the ages of 14-16 were included. Tabata protocol was applied to the HIIT group in addition to their routine training program 3 days/week, for 6 weeks. Erythrocyte deformability was determined using an ectacytometer (LORCA), PV with a rotational viscometer. Total oxidant status (TOS), total antioxidant status (TAS) were measured by kits. RESULTS: HIIT for 6 weeks induced an improvement in performance tests and waist circumference. 6 weeks of HIIT resulted in a decrement, while the last exercise session yielded an increment in RBC deformability. PV and TOS of HIIT groups were decreased on the 6th week. CONCLUSIONS: Our results demonstrate that, HIIT in addition to the routine exercise program is beneficial for improving performance and blood fluidity as well as decreasing oxidative stress in basketball players. Therefore, HIIT seems as an efficient training strategy for highly-trained individuals.
Subject(s)
Basketball , Adolescent , Humans , Basketball/physiology , Oxidative Stress , Antioxidants/metabolism , Exercise/physiology , Erythrocyte Deformability/physiology , OxidantsABSTRACT
Exposure to both oxidative and shear stress, a condition that the red blood cell (RBC) continuously experiences in the circulation in vivo can be mimicked in a Couette type viscometer and monitored by ektacytometry. RBCs maintain their deformation and orientation under shear stress and oxidative stress until a threshold is reached at which these conditions appear to overwhelm the elaborate and complex pathways that maintain a proper redox environment in the cell. Oxidative stress under shear alters the ability of the cell to deform, changes cell morphology, its orientation in the shear stress field, and appears to alter intracellular and membrane characteristics. The application of the RoxyScan technology allows the comparison of oxidant effects and the role of antioxidant systems. This provides the opportunity to study the ability of RBC to deal with oxidative stress in various conditions, including RBC disorders such as sickle cell disease (SCD).
Subject(s)
Anemia, Sickle Cell , Erythrocyte Deformability , Humans , Erythrocytes/metabolism , Oxidative Stress , Oxidation-ReductionABSTRACT
Several diseases like Sickle Cell Anemia, Thalassemia, Hereditary Spherocytosis, Malaria, and Micro-angiopathic Hemolytic Anemia can alter the normal shape of red blood cells (RBCs). The objective of this study is to gain insight into how a change in RBC deformability can affect blood heat transfer. The heat sink effect in a bifurcated vessel with two asymptotic cases (case 1: deformable and case 2: nondeformable RBCs) is being studied during hyperthermia treatment in a three-dimensional bifurcated vessel, whose wall is being subjected to constant heat flux boundary condition. Euler-Euler multiphase method along with the granular model and Kinetic theory is used to include the particle nature of RBCs during blood flow in the current model. To enhance the efficiency of the numerical model, user-defined functions (UDFs) are imported into the model from the C++ interface. The numerical model used is verified with the experimental results from (Carr and Tiruvaloor, 1989, "Enhancement of Heat Transfer in Red Cell Suspensions In Vitro Experiments," ASME J. Biomech. Eng., 111(2), pp. 152-156; Yeleswarapu et al. 1998, "The Flow of Blood in Tubes: Theory and Experiment," Mech. Res. Commun., 25(3), pp. 257-262). The results indicate that the deformability of RBCs can change both the flow dynamics and heat sink effect in a bifurcated vessel, which subsequently affects the efficacy and efficiency of the thermal ablation procedure. Both spatial and transient Nusselt numbers of blood flow with deformable RBCs are slightly higher compared to the one with nondeformable RBCs.
Subject(s)
Hot Temperature , Hyperthermia, Induced , Erythrocyte Deformability/physiology , Erythrocytes , HemodynamicsABSTRACT
Multiple myeloma (MM) is considered to be one of the hematological malignancies formed by excessive and abnormal proliferation of plasmocytes. Among other parameters, several blood tests are used to diagnose multiple myeloma. The hemorheological profile in multiple myeloma is not widely studied. Hemorheology includes the study of measuring the deformability and aggregation of erythrocytes, blood viscosity, and sedimentation rate. The degree of deformability of blood cells is necessary to maintain proper vital functions. Proper deformability of red blood cells ensures proper blood circulation, tissue oxidation and carbon dioxide uptake. The aim of the study was to compare morphology and blood rheology parameters in patients with MM and healthy individuals. The study included 33 patients with MM, and 33 healthy subjects of the same age. The hematological blood parameters were evaluated using ABX MICROS 60 hematology analyzer. The LORCA Analyzer to study erythrocyte aggregation and deformability. Patients with MM had lower red blood cells count (RBC) (9.11%) (p < 0.001) and half time of total aggregation (T1/2) (94.29%) (p < 0.001) values and higher mean corpuscular volume (MCV) (5.50%) (p < 0.001), aggregation index (AI) (68.60%) (p < 0.001), total extent of aggregation (AMP) (87.92%) (p < 0.001) values than the healthy control group. Aggregation in patients with MM is different compared to healthy individuals. It was observed that the percentage of cell aggregation is almost 50% higher than in the control group. The study of morphology, aggregation and deformability of erythrocytes in patients with suspected MM may be helpful in making clinical decisions.
Subject(s)
Erythrocyte Deformability , Multiple Myeloma , Humans , Hemorheology , Erythrocytes , Erythrocyte Aggregation , Rheology , Blood Viscosity , Blood SedimentationABSTRACT
BACKGROUND: The triad "insulin resistance, prediabetes, diabetes" is three independent neologies with characteristic features and development. In addition, each are characterized by progression and the possibility of transition from one form to other. Due to the fact that diabetes is one of the common diseases associated with high rates of disability, it is necessary to improve diagnostic methods and educational regimens for successful prevention and treatment of the disease. OBJECTIVE: We investigated Band 3 protein (B3p) level, osmotic resistance of erythrocytes, the total antioxidant activity (TAA) of blood serum, level of HbA1 in group patients with insulin resistance (IR), prediabetes, and Type 2 Diabetes Mellitus (T2DM) and comparative with health control group. METHODS: We used original, accurate research methods that measure the essence of the studied quantities. RESULTS: Disruptions of glucose and insulin homeostasis ay lead to the initiation of oxidative stress (in our study demonstrated by a decrease of TAA of blood serum) increased redox-sensitive PTP activity and aberrant band 3 phosphorylation, potentially leading to reduced erythrocyte deformability. At the same time glycation of Hb during T2DM may affect its cross-link with membrane proteins, in particular with B3p, and although appears to limit its cross-linking and decrease its clusterization ability, induces alterations in the cytoskeletal matrix, and thereby decrease erythrocytes' osmotic resistance making them more susceptible to hemolysis. CONCLUSIONS: The osmotic resistance of the erythrocytes can be used as a sensitive marker for the detection of the early stages of hyperglycemia (prediabetes). This set of clinical trials will make it possible to identify diseases that make up the triad at an early stage. Early detection of disorders and continued research in this direction will help in the development of a diagnostic scheme for the prevention of such patients. Based on our data, research into anti-oxidation drugs is very important. With the help of the array of studies described in the article and antioxidant treatment, the likelihood of successful treatment will increase.
Subject(s)
Diabetes Mellitus, Type 2 , Erythrocytes , Hyperglycemia , Insulin Resistance , Humans , Diabetes Mellitus, Type 2/blood , Hyperglycemia/blood , Erythrocytes/metabolism , Male , Female , Middle Aged , Oxidative Stress , Erythrocyte Deformability , Anion Exchange Protein 1, Erythrocyte/metabolism , Adult , Prediabetic State/blood , AntioxidantsABSTRACT
BACKGROUND: In the Fontan palliation for single ventricle heart disease (SVHD), pulmonary blood flow is non-pulsatile/passive, low velocity, and low shear, making viscous power loss a critical determinant of cardiac output. The rheologic properties of blood in SVHD patients are essential for understanding and modulating their limited cardiac output and they have not been systematically studied. We hypothesize that viscosity is decreased in single ventricle circulation. METHODS: We evaluated whole blood viscosity, red blood cell (RBC) aggregation, and RBC deformability to evaluate changes in healthy children and SVHD patients. We altered suspending media to understand cellular and plasma differences contributing to rheologic differences. RESULTS: Whole blood viscosity was similar between SVHD and healthy at their native hematocrits, while viscosity was lower at equivalent hematocrits for SVHD patients. RBC deformability is increased, and RBC aggregation is decreased in SVHD patients. Suspending SVHD RBCs in healthy plasma resulted in increased RBC aggregation and suspending healthy RBCs in SVHD plasma resulted in lower RBC aggregation. CONCLUSIONS: Hematocrit corrected blood viscosity is lower in SVHD vs. healthy due to decreased RBC aggregation and higher RBC deformability, a viscous adaptation of blood in patients whose cardiac output is dependent on minimizing viscous power loss. IMPACT: Patients with single ventricle circulation have decreased red blood cell aggregation and increased red blood cell deformability, both of which result in a decrease in blood viscosity across a large shear rate range. Since the unique Fontan circulation has very low-shear and low velocity flow in the pulmonary arteries, blood viscosity plays an increased role in vascular resistance, therefore this work is the first to describe a novel mechanism to target pulmonary vascular resistance as a modifiable risk factor. This is a novel, modifiable risk factor in this patient population.
Subject(s)
Blood Viscosity , Erythrocyte Aggregation , Erythrocyte Deformability , Fontan Procedure , Humans , Child , Heart Defects, Congenital/surgery , Heart Defects, Congenital/physiopathology , Male , Female , Hematocrit , Univentricular Heart/surgery , Univentricular Heart/physiopathology , Child, Preschool , Heart Ventricles/physiopathology , Heart Ventricles/abnormalities , Cardiac Output , Adolescent , ErythrocytesABSTRACT
BACKGROUND: Arteriovenous malformations (AVMs) are vascular anomalies characterized by abnormal shunting between arteries and veins. The progression of the AVMs and their hemodynamic and rheological relations are poorly studied, and there is a lack of a feasible experimental model. OBJECTIVE: To establish a model that cause only minimal micro-rheological alterations, compared to other AV models. METHODS: Sixteen female Sprague Dawley rats were randomly divided into control and AVM groups. End-to-end anastomoses were created between the saphenous veins and arteries to mimic AVM nidus. Hematological and hemorheological parameters were analyzed before surgery and on the 1st, 3rd, 5th, 7th, 9th, and 12th postoperative weeks. RESULTS: Compared to sham-operated Control group the AVM group did not show important alterations in hematological parameters nor in erythrocyte aggregation and deformability. However, slightly increased aggregation and moderately decreased deformability values were found, without significant differences. The changes normalized by the 12th postoperative week. CONCLUSIONS: The presented rat model of a small-caliber AVM created on saphenous vessels does not cause significant micro-rheological changes. The alterations found were most likely related to the acute phase reactions and not to the presence of a small-caliber shunt. The model seems to be suitable for further studies of AVM progression.
Subject(s)
Arteriovenous Malformations , Disease Models, Animal , Rats, Sprague-Dawley , Animals , Rats , Female , Arteriovenous Malformations/pathology , Saphenous Vein/pathology , Hemorheology , Arteriovenous Shunt, Surgical , Erythrocyte Deformability , Erythrocyte AggregationABSTRACT
BACKGROUND: Hemorheological parameters have been reported to be altered in cardiovascular disease. Major depression has been associated with increased risk of cardiovascular disease. OBJECTIVE: Our hypothesis is that hemorheological parameters are disturbed in major depressive disorder. METHODS: Major depressive disorder and control groups consisted of 50 subjects. Plasma viscosity, erythrocyte aggregation, erythrocyte deformability, hematological parameters and hematological parameters were examined. RESULTS: Plasma viscosity was statistically significantly higher, erythrocyte elongation index at 0.53âPa and 0.95âPa was lower, and MCV, MCH, and MCHC values were also lower in the major depression group (Pâ<â0.05). Elongation index and plasma viscosity were correlated with depressive symptomatology. CONCLUSIONS: The increased plasma viscosity and decreased elongation index of erythrocytes indicate an unfavorable hemorheological situation in patients with major depressive disorder compared with healthy controls. The results of this study confirm the findings of studies finding a potential threat to cardiovascular health from major depressive disorder. Increased plasma viscosity and decreased erythrocyte elongation index in depressed patients may be risk factors for cardiovascular events and provide data on the causality of the association between depression and cardiovascular disease.
Subject(s)
Blood Viscosity , Depressive Disorder, Major , Erythrocyte Aggregation , Erythrocyte Deformability , Hemorheology , Humans , Depressive Disorder, Major/blood , Male , Female , Adult , Middle Aged , Cardiovascular Diseases/bloodABSTRACT
Deterministic Lateral Displacement (DLD) device has gained widespread recognition and trusted for filtering blood cells. However, there remains a crucial need to explore the complex interplay between deformable cells and flow within the DLD device to improve its design. This paper presents an approach utilizing a mesoscopic cell-level numerical model based on dissipative particle dynamics to effectively capture this complex phenomenon. To establish the model's credibility, a series of numerical simulations were conducted and the numerical results were validated with nominal experimental data from the literature. These include single cell stretching experiment, comparisons of the morphological characteristics of cells in DLD, and comparison the specific row-shift fraction of DLD required to initiate the zigzag mode. Additionally, we investigate the effect of cell rigidity, which serves as an indicator of cell health, on average flow velocity, trajectory, and asphericity. Moreover, we extend the existing theory of predicting zigzag mode for solid spherical particles to encompass the behavior of red blood cells. To achieve this, we introduce a new concept of effective diameter and demonstrate its applicability in providing highly accurate predictions across a wide range of conditions.
Subject(s)
Erythrocyte Deformability , Erythrocytes , FiltrationABSTRACT
The intrinsic physical and mechanical properties of red blood cells (RBCs), including their geometric and rheological characteristics, can undergo changes in various circulatory and metabolic diseases. However, clinical diagnosis using RBC biophysical phenotypes remains impractical due to the unique biconcave shape, remarkable deformability, and high heterogeneity within different subpopulations. Here, we combine the hydrodynamic mechanisms of fluid-cell interactions in micro circular tubes with a machine learning method to develop a relatively high-throughput microfluidic technology that can accurately measure the shear modulus of the membrane, viscosity, surface area, and volume of individual RBCs. The present method can detect the subtle changes of mechanical properties in various RBC components at continuum scales in response to different doses of cytoskeletal drugs. We also investigate the correlation between glycosylated hemoglobin and RBC mechanical properties. Our study develops a methodology that combines microfluidic technology and machine learning to explore the material properties of cells based on fluid-cell interactions. This approach holds promise in offering novel label-free single-cell-assay-based biophysical markers for RBCs, thereby enhancing the potential for more robust disease diagnosis.
Subject(s)
Erythrocyte Deformability , Erythrocytes , Viscosity , Rheology , Microfluidics/methodsABSTRACT
BACKGROUND: Even though cardiovascular stenting is widely used for the treatment of coronary artery disease, information on how it can affect the hematological and hemorheological profile is scarce in the literature. Most of the work on this issue is based on theoretical or computational fluid dynamics models, lacking in-depth in vitro and in vivo experimental verification. OBJECTIVE: This work investigates, in an in vivo setting, the effects of stenting and the implantation time-course on hematological and hemorheological parameters that could potentially compromise the device's functionality and longevity. METHODS: Custom-made self-expanding nitinol stents were implanted in the common carotid artery of male CD1 mice. Whole blood samples were collected from control (non-stented) and stented animals at 5 and 10 weeks post-implantation. Hematological measurements and blood viscosity, red blood cell aggregation, and deformability were performed using standard techniques. RESULTS: Implant-induced changes were observed in some of the hematological and hemorheological indices. Blood viscosity seems to have been negatively affected by an increased hematocrit and reduced RBC deformability, at 10 weeks post-implantation, despite a slight decrease in RBC aggregation. CONCLUSIONS: Although the alterations observed may be the result of the peri-implant inflammatory response, the physiological consequences due to hemorheological changes need to be further investigated.
Subject(s)
Blood Viscosity , Hemorheology , Stents , Animals , Mice , Male , Erythrocyte Deformability , Erythrocyte Aggregation , Alloys , HematocritABSTRACT
The complex, systemic pathology of sickle cell disease is driven by multiple mechanisms including red blood cells (RBCs) stiffened by polymerized fibers of deoxygenated sickle hemoglobin. A critical step toward understanding the pathologic role of polymer-containing RBCs is quantifying the biophysical changes in these cells in physiologically relevant oxygen environments. We have developed a microfluidic platform capable of simultaneously measuring single RBC deformability and oxygen saturation under controlled oxygen and shear stress. We found that RBCs with detectable amounts of polymer have decreased oxygen affinity and decreased deformability. Surprisingly, the deformability of the polymer-containing cells is oxygen-independent, while the fraction of these cells increases as oxygen decreases. We also find that some fraction of these cells is present at most physiologic oxygen tensions, suggesting a role for these cells in the systemic pathologies. Additionally, the ability to measure these pathological cells should provide clearer targets for evaluating therapies.
Subject(s)
Anemia, Sickle Cell , Oxygen Saturation , Humans , Erythrocytes , Erythrocyte Deformability , Polymers , OxygenABSTRACT
Long-chain polyunsaturated fatty acids (LC-PUFAs) are important modulators of red blood cell (RBC) rheology. Dietary LC-PUFAs are readily incorporated into the RBC membrane, improving RBC deformability, fluidity, and hydration. Female C57BL/6J mice consumed diets containing increasing amounts of fish oil (FO) ad libitum for 8 weeks. RBC deformability, filterability, and post-transfusion recovery (PTR) were evaluated before and after cold storage. Lipidomics and lipid peroxidation markers were evaluated in fresh and stored RBCs. High-dose dietary FO (50%, 100%) was associated with a reduction in RBC quality (i.e., in vivo lifespan, deformability, lipid peroxidation) along with a reduced 24 h PTR after cold storage. Low-dose dietary FO (6.25-12.5%) improved the filterability of fresh RBCs and reduced the lipid peroxidation of cold-stored RBCs. Although low doses of FO improved RBC deformability and reduced oxidative stress, no improvement was observed for the PTR of stored RBCs. The improvement in RBC deformability observed with low-dose FO supplementation could potentially benefit endurance athletes and patients with conditions resulting from reduced perfusion, such as peripheral vascular disease.
Subject(s)
Dietary Fats, Unsaturated , Erythrocyte Deformability , Humans , Female , Mice , Animals , Mice, Inbred C57BL , Erythrocytes/metabolism , Fish Oils/pharmacology , Fish Oils/metabolism , Fatty Acids, Unsaturated/metabolism , Fatty Acids/metabolism , Dietary Fats, Unsaturated/metabolism , Blood Preservation/methodsABSTRACT
In routine hematological instruments, blood cells are counted and sized by monitoring the impedance signals induced during their passage through a Coulter orifice. However, only signals associated with centered paths in the aperture are considered for analysis, while the rejected measurements, caused by near-wall trajectories, can provide additional information on red blood cells (RBC), as recent publications suggest. To assess usefulness of two new parameters in describing alterations in RBC properties, we performed a pilot study to compare blood samples from patients with diabetes mellitus (DM), frequent pathological condition associated with impairment in RBC deformability, versus controls. A total of 345 blood samples were analyzed: 225 in the DM group and 120 in the control group. A diagram of [Formula: see text] and [Formula: see text], the two new parameters derived from the analysis of impedancemetry pulses, was used to compare distribution of RBC subpopulations between groups. To discriminate RBC from DM and control individuals, based on our multiparametric analysis, we built a score from variables derived from [Formula: see text] matrix which showed good performances: area under the receiving operating characteristic curve 0.948 (0.920-0.970), p<0.0001; best discriminating value: negative predictive value 94.7%, positive predictive value was 78.4%. These results seem promising to approach RBC alterations in routine laboratory practice. The related potential clinically relevant outcomes remain to be investigated.