Microfluidic Characterization of Red Blood Cells Microcirculation under Oxidative Stress

Microcirculation is one of the basic functional processes where the main gas exchange between red blood cells (RBCs) and surrounding tissues occurs. It is greatly influenced by the shape and deformability of RBCs, which can be affected by oxidative stress induced by different drugs and diseases leading to anemia. Here we investigated how in vitro microfluidic characterization of RBCs transit velocity in microcapillaries can indicate cells damage and its correlation with clinical hematological analysis. For this purpose, we compared an SU-8 mold with an Si-etched mold for fabrication of PDMS microfluidic devices and quantitatively figured out that oxidative stress induced by tert-Butyl hydroperoxide splits all RBCs into two subpopulations of normal and slow cells according to their transit velocity. Obtained results agree with the hematological analysis showing that such changes in RBCs velocities are due to violations of shape, volume, and increased heterogeneity of the cells. These data show that characterization of RBCs transport in microfluidic devices can directly reveal violations of microcirculation caused by oxidative stress. Therefore, it can be used for characterization of the ability of RBCs to move in microcapillaries, estimating possible side effects of cancer chemotherapy, and predicting the risk of anemia.

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Damage of erythrocytes and blood plasma macromolecules in patients with alcoholism and the membrane-protective effect of lithium salts

ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia» ◽

10.25557/0031-2991.2021.02.22-29 ◽

2021 ◽

pp. 22-29

Author(s):

Т.П. Ветлугина ◽

В.Д. Прокопьева ◽

Е.В. Плотников ◽

Е.Г. Ярыгина ◽

В.Ф. Лебедева ◽

...

Keyword(s):

Oxidative Stress ◽

Protective Effect ◽

Red Blood Cells ◽

Blood Cells ◽

Oxidative Modification ◽

Toxic Effects ◽

Lithium Salts ◽

Hematological Analysis ◽

Spectrum Disorders

Введение. Нарушения параметров красной крови при алкоголизме, развитие окислительного стресса с увеличением продуктов окислительной модификации биомолекул обусловливают актуальность поиска защитных средств эритроцитов от токсического воздействия этанола. Перспективными для такого поиска могут быть соли лития, учитывая применение препаратов лития в качестве нормотимиков для лечения расстройств аффективного спектра. Цель исследования - изучение показателей красной крови, факторов окислительного стресса у больных алкоголизмом и оценка защитного потенциала солей лития (аскорбата, сукцината, пирувата, фумарата и карбоната) от токсического действия этанола на эритроциты. Методика. Материалом для исследования служили образцы крови 59 мужчин больных алкоголизмом и 38 здоровых мужчин (контроль). Гематологический анализ крови осуществляли на гематологическом анализаторе Micros 60. В плазме крови определяли показатели окислительного повреждения белков (по уровню карбонилированных белков) и перекисного окисления липидов (по уровню ТБК-реактивных продуктов) спектрофотометрическим методом. Мембранопротекторный эффект солей лития оценивали в опытах in vitro по степени гемолиза эритроцитов при действии этанола в присутствии солей лития в концентрации 1,2 ммоль/л в расчете на ион лития. Результаты. У больных алкоголизмом установлено повышение среднего объема эритроцитов (MCV) и снижение параметров остальных показателей красной крови. Концентрации карбонилированных белков и ТБК-реактивных продуктов в плазме крови пациентов превышали контрольные значения. Соли лития (фумарат, пируват и аскорбат) в 1,5-1,7 раз повышали устойчивость эритроцитов к гемолизу, индуцированному этанолом. Карбонат лития такого эффекта не оказывал. Заключение. У больных алкоголизмом обнаружены существенные повреждения эритроцитов и макромолекул плазмы крови. Фумарат, пируват и аскорбат лития демонстрировали высокий мембранопротекторный эффект, повышая устойчивость эритроцитов к токсическому действию этанола. Данные соли лития расширяют арсенал мембранопротекторных соединений, что в сочетании с нормотимическим действием лития позволяет использовать их для разработки импортозамещающих лекарственных средств лечения расстройств аффективного спектра. Introduction. In alcoholism, red blood variables are altered, and oxidative stress, along with an increase in the oxidative modification of biomolecules products, occurs. These effects have stimulated the search for agents to protect red blood cells against the toxic effects of ethanol. Lithium salts may be promising in this regard, since lithium preparations are used as normothymics for treating affective spectrum disorders. The aim of this study was to investigate red blood indexes, factors of oxidative stress in patients with alcoholism and to assess the protective potential of the lithium salts, ascorbate, succinate, pyruvate, fumarate and carbonate, against the toxic effect of ethanol on red blood cells. Methods. Blood samples were collected from 59 alcoholic men and 38 healthy men. Hematological analysis was performed with a Micros 60 hematological analyzer. In plasma, oxidative damage of proteins was determined by the amount of carbonylated proteins (CP), and lipid peroxidation was determined by the amount of TBA-reactive substances (TBARS), as measured spectrophotometrically. The membrane-protective effect of lithium salts was evaluated in vitro by the degree of hemolysis of red blood cells treated with ethanol in the presence of lithium salts at a concentration of 1.2 mmol/l lithium ion. Results. In alcoholic patients, compared with normal patients, an increase in the mean corpuscular volume (MCV) and a decrease in all other red cell variables were found. Concentrations of CP and TBARS in plasma exceeded normal values. The lithium salts, fumarate, pyruvate, and ascorbate, elevated the resistance of red blood cells to ethanol-induced hemolysis by 1.5-1.7 times. Lithium carbonate had no effect. Conclusion. Significant damage to red blood cells and plasma macromolecules was found in alcoholic patients. Lithium fumarate, pyruvate and ascorbate had a large membrane-protective effect. Thus, these salts increased the resistance of red blood cells to the toxic effects of ethanol. These salts expand the arsenal of membrane-protective compounds, which in combination with the normothymic effect of lithium, will allow them to be used in developing drugs for treatment of affective spectrum disorders.

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In vitro characterization of engineered red blood cells as viral traps against HIV-1 and SARS-CoV-2

Molecular Therapy — Methods & Clinical Development ◽

10.1016/j.omtm.2021.03.003 ◽

2021 ◽

Vol 21 ◽

pp. 161-170 ◽

Cited By ~ 1

Author(s):

Magnus A.G. Hoffmann ◽

Collin Kieffer ◽

Pamela J. Bjorkman

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

In Vitro Characterization ◽

Hiv 1

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In Vitro Assay for Single-cell Characterization of Impaired Deformability in Red Blood Cells under Recurrent Episodes of Hypoxia

Lab on a Chip ◽

10.1039/d1lc00598g ◽

2021 ◽

Author(s):

YUHAO QIANG ◽

Jia Liu ◽

Ming Dao ◽

E Du

Keyword(s):

Shear Stress ◽

Red Blood Cells ◽

Single Cell ◽

Blood Cells ◽

Blood Circulation ◽

In Vitro Assay ◽

Vitro Assay ◽

Cyclic Shear

Red blood cells (RBCs) are subjected to recurrent changes in shear stress and oxygen tension during blood circulation. The cyclic shear stress has been identified as an important factor that...

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Increased susceptibility of microcytic red blood cells to in vitro oxidative stress

European Journal Of Haematology ◽

10.1111/j.1600-0609.1995.tb00705.x ◽

2009 ◽

Vol 55 (5) ◽

pp. 327-331 ◽

Cited By ~ 51

Author(s):

J. L. Vives Corrons ◽

A. Miguel-Garcia ◽

M. A. Pujades ◽

A. Miguel-Sosa ◽

S. Cambiazzo ◽

...

Keyword(s):

Oxidative Stress ◽

Red Blood Cells ◽

Blood Cells ◽

Increased Susceptibility

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Oxidative Stress and Increased Destruction of Red Blood Cells Contribute to the Pathophysiology of Anemia Caused By DMT1 Deficiency

Blood ◽

10.1182/blood.v124.21.4027.4027 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4027-4027 ◽

Cited By ~ 1

Author(s):

Zuzana Zidova ◽

Daniel Garcia-Santos ◽

Katarina Kapralova ◽

Pavla Koralkova ◽

Renata Mojzikova ◽

...

Keyword(s):

Oxidative Stress ◽

Red Blood Cells ◽

Blood Cells ◽

Fetal Liver ◽

Erythroid Differentiation ◽

Heme Oxygenase 1 ◽

Wild Type ◽

Divalent Metal Transporter 1 ◽

Oxidative Defense

Abstract Inactivating mutations in divalent metal transporter 1 (DMT1) are associated with a severe defect in erythroid iron utilization and cause moderate to severe hypochromic microcytic anemia in human patients and two rodent models. We have previously shown that DMT1 deficiency impairs erythroid differentiation, induces apoptosis of erythroid precursors and causes the suppression of colony-forming capacity of erythroid progenitors. Using in vitro cultures of fetal liver cells we were able to recapitulate this in vivo defect. We confirmed abnormal pattern of erythroid differentiation and increased apoptosis (2.5-times) of DMT1-mutant erythroblasts when compared to wild-type (wt) fetal liver erythroblats. Determination of 2’,7’-Dichlorofluorescein diacetate-dependent intensity of fluorescence, which is proportional to the concentration of reactive oxygen species (ROS), revealed elevated levels of ROS in DMT1-mutant erythroblats when compared to wt erythroblast. This result suggests that oxidative stress contributes to the apoptosis in DMT1-mutant cells. We also observed that the defective erythroid differentiation of DMT1-mutant erythroblasts is marked by a blunted induction of heme oxygenase-1, an enzyme that co-regulates erythroid differentiation by controlling the heme regulatory pool in erythroid cells (Garcia-Santos et al., Blood, 2014, 123 (14): 2269-77). In further studies we focused on mature red blood cells (RBC), because it is known that nutritional iron deficiency and certain types of congenital hypochromic anemia are associated with increased levels of ROS and shortened life span of RBC that can be at least partially attributed to a programmed cell death of erythrocytes, so called eryptosis (Lang et al., Int J Biochem Cell Biol, 2012, 44 (8): 1236-43). Using labeling with carboxyfluorescein diacetate succinimidyl ester, we observed an accelerated clearance of DMT1-mutant RBC from circulating blood when compared to wild-type RBC. In vitro, DMT1-mutant RBC exposed to hyperosmotic shock or glucose depletion showed significantly increased levels of phosphatidylserine on the membrane detected by Annexin V binding. Together, these results confirmed eryptosis of DMT1-mutant RBC. As eryptosis is proposed to be triggered via activation of Ca2+ cation channels, we next measured the concentration of cytosolic Ca2+ using Fluo3/AM fluorescent dye and found significantly elevated content of intracellular Ca2+ in DMT1-mutant RBC when compared to wt RBC. In addition, DMT1-mutant RBC had higher levels of ROS than wt RBC despite significantly increased activity of anti-oxidative defense enzymes; glutathione peroxidase (1.6-times), catalase (1.9-times) and methemoglobin reductase (1.9-times). This indicates that exaggerated anti-oxidative defense in DMT1-mutant RBC is not sufficient to eliminate ROS effectively. Furthermore, DMT1-mutant RBC also showed accelerated anaerobic glycolysis as detected by increased activities of hexokinase (2.5-times), pyruvate kinase (2.4-times), glucose-phosphate isomerase (3.2-times). This result together with reduced ATP/ADP (1.6-times) ratio in DMT1-mutant RBC when compared to wt RBC suggests an increased demand for ATP in DMT1-mutant erythrocytes. In conclusion we propose that increased oxidative stress and accelerated destruction of RBC contribute to the pathophysiology of anemia caused by DMT1-deficiency. Grant support: Czech Grant Agency, grant No. P305/11/1745; Ministry of Health Czech Republic, grant No. NT13587, Education for Competitiveness Operational Program, CZ.1.07/2.3.00/20.0164, Internal Grant of Palacky University Olomouc, LF_2014_011 and in part by the Canadian Institutes of Health Research (D.G-S., P.P.). Disclosures No relevant conflicts of interest to declare.

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Antioxidant-Enhancing Property of the Polar Fraction of Mangosteen Pericarp Extract and Evaluation of Its Safety in Humans

Oxidative Medicine and Cellular Longevity ◽

10.1155/2016/1293036 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 6

Author(s):

Wichit Suthammarak ◽

Pornpayom Numpraphrut ◽

Ratiya Charoensakdi ◽

Neelobol Neungton ◽

Vachara Tunrungruangtavee ◽

...

Keyword(s):

Side Effects ◽

Oral Administration ◽

Red Blood Cells ◽

Blood Cells ◽

Blood Chemistry ◽

Antioxidant Property ◽

Polar Fraction ◽

Cytotoxic Activities

Crude extract from the pericarp of the mangosteen (mangosteen extract [ME]) has exhibited several medicinal properties in both animal models and human cell lines. Interestingly, the cytotoxic activities were always observed in nonpolar fraction of the extract whereas the potent antioxidant was often found in polar fraction. Although it has been demonstrated that the polar fraction of ME exhibited the antioxidant activity, the safety of the polar fraction of ME has never been thoroughly investigated in humans. In this study, we investigated the safety of oral administration of the polar fraction of ME in 11 healthy Thai volunteers. During a 24-week period of the study, only minor and tolerable side effects were reported; no serious side effects were documented. Blood chemistry studies also showed no liver damage or kidney dysfunction in all subjects. We also demonstrated antioxidant property of the polar fraction of ME bothin vitroandin vivo. Interestingly, oral administration of the polar fraction of ME enhanced the antioxidant capability of red blood cells and decreased oxidative damage to proteins within red blood cells and whole blood.

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Исследование транспорта эритроцитов через микроканалы при индукции окислительного стресса трет-бутилпероксидом

Журнал технической физики ◽

10.21883/jtf.2020.09.49689.403-19 ◽

2020 ◽

Vol 90 (9) ◽

pp. 1553

Author(s):

Е.А. Скверчинская ◽

О.Д. Тапинова ◽

Н.А. Филатов ◽

Н.А. Беседина ◽

И.В. Миндукшев ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Membrane ◽

Red Blood Cells ◽

Microfluidic Device ◽

Blood Cells ◽

Butyl Hydroperoxide ◽

Blood Microcirculation ◽

Tert Butyl Hydroperoxide ◽

Microfluidic Analysis ◽

Cytological Evaluation

To investigate the influence of oxidative stress on red blood cells (RBCs) transport in microvessels and capillaries of the blood microcirculation system, we studied their migration in narrow microchannels in a microfluidic device. We measured cells' velocities in the microchannels under the oxidative stress induced by tert-Butyl hydroperoxide at different concentrations and compared the results with a cytological evaluation of the cell membrane transformations. Oxidative stress caused changes in cells’ shape and volume, travelling velocity of the cells decreased and occlusions of microchannels occurred. The developed microfluidic device provided an ability to study microreology of RBCs under the action of hydroperoxide. In the future, such microfluidic analysis may be used to determine the changes in microrheology of RBCs from patients under the influence of xenobiotics.

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Protective Effect of Pitao (Pitavia punctata (R. & P.) Molina) Polyphenols against the Red Blood Cells Lipoperoxidation and the In Vitro LDL Oxidation

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/1049234 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 2

Author(s):

Ricardo I. Castro ◽

Oscar Forero-Doria ◽

Luis Soto-Cerda ◽

A. Peña-Neira ◽

Luis Guzmán

Keyword(s):

Oxidative Stress ◽

Red Blood Cells ◽

Blood Cells ◽

Hypochlorous Acid ◽

Low Density Lipoprotein ◽

Membrane Integrity ◽

Density Lipoprotein ◽

Ldl Oxidation ◽

Quercetin Derivatives

The oxidative stress is characterized by an imbalance between the oxidizing agents and antioxidants; meanwhile, the consumption of antioxidants has been considered as an important tool in the prevention of oxidative stress and its consequences. Pitavia punctata (R. & P.) Molina is an endemic arboreal species from the Chilean Coast Range, in which a large amount of flavonoids has been described. This work focused on characterizing and evaluating, in human erythrocytes, the antioxidant capacity and membrane protection of P. punctata extracts and the in vitro protection of the oxidation of the Low Density Lipoprotein (LDL). The phytochemical screening revealed the presence of Quercetin derivatives and flavonoids, such as (-)-Epicatechin, Kaempferol, and derivatives. The methanolic extract presented an important antioxidant activity, protecting the membrane integrity of the red blood cells against the oxidative damage caused by Hypochlorous acid and inhibiting the oxidation of the LDL lipoprotein.

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