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Cyclic arginine-glycine-aspartic acid-modified red blood cells for drug delivery:Synthesis and in vitro evaluation

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Journal of Pharmaceutical Analysis 2022年第2期12卷 324-331页

作者： Chen Wang Min Wang Yan Zhang Hongxin Jia Binbin Chen Department of Pharmaceutics School of PharmacyXiamen Medical CollegeXiamenFujian361023China Institute of Respiratory Diseases Xiamen Medical CollegeXiamenFujian361023China Department of Pharmaceutics School of PharmacyHarbin University of CommerceHarbin150076China Department of Pharmacy Xiamen Xianyue HospitalXiamenFujian361012China

red blood cells(RBCs)are an excellent choice for cell preparation research because of their biocompatibility,high drug loading,and long half-life.In this study,doxorubicin(DOX)was encapsulated with RBCs as the carrier.The biotin-avidin system binding principle was used to modify biotinylated cyclic arginine-glycine-aspartic acid(cRGD)onto RBC surfaces for accurate targeting,high drug loading,and sustained drug release.The RBC drug delivery system(DDS)was characterized,and the concentration of surface sulfur in the energy spectrum was 6.330%.The physical and chemical properties of RBC DDS were as follows:drug content,0.857 mg/mL;particle size,3339 nm;potential value,12.5 mV;and cumulative release rate,81.35%.There was no significant change in RBC morphology for up to seven days.The results of the targeting and cytotoxicity studies of RBC DDS showed that many RBCs covered the surfaces of U251 cells,and the fluorescence intensity was higher than that of MCF-7 cells.The IC50 value of unmodified drug-loaded RBCs was 2.5 times higher than that of targeted modified drug-loaded RBCs,indicating that the targeting of cancer cells produced satisfactory inhibition.This study confirms that the RBC DDS has the characteristics of accurate targeting,high drug loading,and slow drug release,which increases its likelihood of becoming a clinical cancer treatment in the future.

关键词： Cytotoxicity red blood cells Drug delivery Targeting

Antibody-engineered red blood cell interface for high-performance capture and release of circulating tumor cells

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Bioactive Materials 2022年第5期7卷 32-40页

作者： Haicong Shen Rui Su Jiao Peng Lin Zhu Kunyue Deng Qi Niu Yanling Song Liu Yang Lingling Wu Zhi Zhu Chaoyong Yang MOE Key Laboratory of Spectrochemical Analysis&Instrumentation Collaborative Innovation Center of Chemistry for Energy MaterialsKey Laboratory for Chemical Biology of Fujian ProvinceState Key Laboratory of Physical Chemistry of Solid SurfacesDepartment of Chemical BiologyCollege of Chemistry and EngineeringXiamen UniversityXiamen361005China Department of Hematology Clinical LaboratoryThe First Affiliated Hospital of Xiamen UniversityXiamen361005China Institute of Molecular Medicine Department of Gastrointestinal SurgeryRenji HospitalSchool of MedicineShanghai Jiao Tong University Shanghai200127China

Circulating tumor cells(CTCs),as important liquid biopsy target,can provide valuable information for cancer progress monitoring and individualized treatment.However,current isolation platforms incapable of balancing capture efficiency,specificity,cell viability,and gentle release have restricted the clinical applications of CTCs.Herein,inspired by the structure and functional merits of natural membrane interfaces,we established an antibody-engineered red blood cell(RBC-Ab)affinity interface on microfluidic chip for high-performance isolation and release of CTCs.The lateral fluidity,pliability,and anti-adhesion property of the RBC microfluidic interface enabled efficient CTCs capture(96.5%),high CTCs viability(96.1%),and high CTCs purity(average 4.2-log depletion of leukocytes).More importantly,selective lysis of RBCs by simply changing the salt concentration was utilized to destroy the affinity interface for efficient and gentle release of CTCs without nucleic acid contamination.Using this chip,CTCs were successfully detected in colon cancer samples with 90%sensitivity and 100%specificity(20 patients and 10 healthy individuals).After the release process,KRAS gene mutations of CTCs were identified from all the 5 cancer samples,which was consistent with the results of tissue biopsy.We expect this RBC interface strategy will inspire further biomimetic interface construction for rare cell analysis.

关键词： red blood cells Circulating tumor cells Colon cancer Biomimetic interface

Exact Singularity Subtraction from Boundary Integral Equations in Modeling Vesicles and red blood cells

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Numerical Mathematics(Theory,Methods and Applications) 2014年第4期7卷 413-434页

作者： Alexander Farutin Chaouqi Misbah UniversitéGrenoble I/CNRS Laboratoire Interdisciplinaire de Physique/UMR5588Grenoble F-38041France

The study of vesicles,capsules and red blood cells(RBCs)under flow is a field of active research,belonging to the general problematic of fluid/structure interactions.Here,we are interested in modeling vesicles,capsules and RBCs using a boundary integral formulation,and focus on exact singularity subtractions of the kernel of the integral equations in 3D.In order to increase the precision of singular and near-singular integration,we propose here a refinement procedure in the vicinity of the pole of the Green-Oseen kernel.The refinement is performed homogeneously everywhere on the source surface in order to reuse the additional quadrature nodes when calculating boundary integrals in multiple target points.We also introduce a multi-level look-up algorithm in order to select the additional quadrature nodes in vicinity of the pole of the Green-Oseen kernel.The expected convergence rate of the proposed algorithm is of order δ(1/N^(2))while the computational complexity is of order δ(N^(2) lnN),where N is the number of degrees of freedom used for surface discretization.Several numerical tests are presented to demonstrate the convergence and the efficiency of the method.

关键词： Stokes flow fluid structure interaction boundary integral method red blood cells singularity subtraction

SARS-CoV-2 infection and red blood cells:Implications for long term symptoms during exercise

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Sports Medicine and Health Science 2021年第3期3卷 181-182页

作者： Francesco Misiti Department of Human Sciences Society and HealthUniversity of Cassino and Southern LazioV.S.AngeloLoc.Folcara03043CassinoFRItaly

Skeletal muscle blood flow is coupled with the oxygenation state of hemoglobin,whereby the red blood cell(RBC)functions as an oxygen sensor and releases adenosine triphosphate(ATP)during deoxygenation to evoke vasodilation,an increase in blood flow and O2 delivery to tis-sues,presumably acting on endothelial cell purinergic receptors.1 A signal transduction pathway relating deoxygenation to ATP release has been described and includes band 3,the most abundant membrane protein in mature RBCs,the heterotrimeric G protein Gi and adenylyl cyclase(AC).

关键词： red blood cells Exercise COVID-19 Metabolism

Water Efflux in red blood cells of Sickle Cell Patients under Spontaneous Deoxygenation

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Journal of Biosciences and Medicines 2016年第12期4卷 152-161页

作者： Manuel Arsenio Lores Guevara Yulianela Mengana Torres Juan Carlos García Naranjo Andrés Ramírez Aguilera Lidia Clara Suárez Beyrio María Altagracia Marichal Felue Teresa Simón Brada Jan Philippé Centro de Biofí sica Mé dica Santiago de Cuba Cuba Hospital General “Juan Bruno Zayas Alfonso” Santiago de Cuba Cuba Department of Clinical Chemistry Immunology and Microbiology Ghent University Ghent Belgium

The water transport through red blood cells (RBC) membrane has been previously studied in Sickle Cell Disease (SCD) using oxygenated RBC or under complete deoxygenation. In this work, the water efflux in RBC of sickle cell patients was studied under spontaneous deoxygenation conditions. With that purpose, a magnetic resonance method was used to evaluate the water exchange time (τe) and the permeability through the erythrocyte membrane (P) measuring the spin-spin relaxation time (T2) in doped and non-doped RBC. Carr-Purcell-Meiboon-Gill (CPMG) pulse sequence was used to measure T2 in a magnetic resonance console coupled to one homogeneous magnet system (0.095 T). An increase of the water transport in RBC from sickle cell patients was observed and characterized with a τe value of 15.2 ± 0.8 ms. The abnormal activation of the Psickle, Gardos, and potassium chloride cotransporter channels starting from deoxygenation, as well as, the possible appearance of new pores due to the increase of the hemoglobin-membrane interaction, are suggested to explain this abnormal transport phenotype. The change of the water volume to surface ratio (V/S) in the sickle cells is also suggested to be considered in P calculation under deoxygenation. The results obtained in this work increase the fundamental knowledge about molecular mechanism involved in SCD and could be useful in the development of new methods for diagnostic and treatment evaluation.

关键词： red blood cells Water Permeability Nuclear Magnetic Relaxation Sickle Cell Disease

Improving Oxygen Binding of Desiccated Human red blood cells

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Advances in Bioscience and Biotechnology 2016年第2期7卷 47-54页

作者： Steingrimur Stefansson David S. Chung Jamie Yoon Won Seok Yoo Young Wook Park George Kim David Hahn Huyen Le Sung-Jae Chung Stephen P. Bruttig David H. Ho HeMemics Biotechnologies Inc. Rockville MD USA Division of Biology & Medicine Brown University Providence RI USA Earl Warren College UC San Diego CA USA Department of Chemistry Michigan University Ann Arbor MI USA Seoul International School Seongnam South Korea Fuzbien Technology Institute Rockville MD USA Nauah Solutions Mclean VA USA Marymount University School of Arts & Science Arlington VA USA

Desiccating human red blood cells (RBCs) to increase their storage life has been the subject of intense research for a number of years. However, drying RBCs invariably compromises their integrity and has detrimental effects on hemoglobin function due to autoxidation. We have previously demonstrated an RBC desiccation and rehydration process that preserves RBC antigenic epitopes better than frozen RBCs. This study expands on those observations by examining what effects this desiccation process has on RBC hemoglobin function with respect to oxygen binding properties. In this paper, we examined RBCs from normal donors which were desiccated to 25% moisture content and stored dry for 2 weeks at room temperature prior to rehydration with plasma followed by structural and functional studies. Our data showed that approximately 98% of the RBCs were intact upon rehydration based on hemolysis assays. Oxygen dissociation curves for the desiccated/rehydrated RBCs showed a left shift compared to fresh RBCs (pO2 = 17 mmHg vs. 26 mmHg, respectively). The desiccated/rehydrated RBCs also showed an increase in methemoglobin compared to fresh RBCs (4.5% vs 0.9%, respectively). 2,3-Diphosphoglycerate concentration of the desiccated/rehydrated RBCs was reduced by 20%. In conclusion, although this RBC dehydration process preserves RBC integrity and hemoglobin oxygen binding properties better than most other dehydration techniques described so far, further optimization and long-term studies are needed to make this procedure acceptable for human transfusion.

关键词： Desiccation red blood cells Oxygen Binding Hemolysis Methemoglobin 2 3-Diphosphoglycerate

red blood cells (ABCs) in plasma can survive freeze thaw

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中国输血杂志 2001年第S1期14卷 422-页

Multiparametric characterization of red blood cell physiology after hypotonic dialysis based drug encapsulation process

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Acta Pharmaceutica Sinica B 2022年第4期12卷 2089-2102页

作者： Mélanie Robert Bastien Laperrousaz Diana Piedrahita Emilie-Fleur Gautier Travis Nemkov Florian Dupuy Elie Nader Virginie Salnot Patrick Mayeux Angelo D’Alessandro Catherine Lavazec Philippe Joly Alexander Scheer Philippe Connes Agnès Cibiel Erytech Pharma Lyon 69008France Laboratoire interuniversitaire de Biologie de la Motricité(LIBM)EA7424 Team«Vascular Biology and Red Blood Cell»Universite´Claude Bernard Lyon 1Universite´de LyonLyon 69008France Labex GR-Ex Paris 75015France 3P5 proteom’IC facility Universitéde ParisInstitut CochinINSERMCNRSParis 75014France University of Colorado Anschutz Medical CampusAuroraCO 80045USA Omix Technologies Inc.AuroraCO 80045USA Inserm U1016 CNRS 8104Universitéde ParisInstitut CochinParis 75014France Laboratoire de Biochimie et de Biologie Moléculaire UF de biochimie des pathologies e´rythrocytairesCentre de Biologie et de Pathologie EstHospices Civils de LyonBron 69500France

red blood cells(RBCs)can act as carriers for therapeutic agents and can substantially improve the safety,pharmacokinetics,and pharmacodynamics of many drugs.Maintaining RBCs integrity and lifespan is important for the efficacy of RBCs as drug carrier.We investigated the impact of drug encapsulation by hypotonic dialysis on RBCs physiology and integrity.Several parameters were compared between processed RBCs loaded with l-asparaginase(“eryaspase”),processed RBCs without drug and non-processed RBCs.Processed RBCs were less hydrated and displayed a reduction of intracellular content.We observed a change in the metabolomic but not in the proteomic profile of processed RBCs.Encapsulation process caused moderate morphological changes and was accompanied by an increase of RBCs-derived Extracellular Vesicles release.Despite a decrease in deformability,processed RBCs were not mechanically retained in a spleen-mimicking device and had increased surface-to-volume ratio and osmotic resistance.Processed RBCs half-life was not significantly affected in a mouse model and our previous phase 1 clinical study showed that encapsulation of asparaginase in RBCs prolonged its in vivo half-life compared to free forms.Our study demonstrated that encapsulation by hypotonic dialysis may affect certain characteristics of RBCs but does not significantly affect the in vivo longevity of RBCs or their drug carrier function.

关键词： red blood cells Drug carrier Hypotonic dialysis L-Asparaginase Omics Rheology Morphology Senescence markers

Characterization of the interaction of nanobubble ultrasound contrast agents with human blood components

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生物活性材料(英文) 2023年第1期19卷 642-652页

作者： Michaela B.Cooley Eric C.Abenojar Dana Wegierak Anirban Sen Gupta Michael C.Kolios Agata A.Exner Department of Biomedical Engineering Case Western Reserve UniversityClevelandOH44106United States Department of Radiology Case Western Reserve UniversityClevelandOH44106United States Department of Physics Ryerson UniversityTorontoOntarioM5B 2K3Canada Institute for Biomedical Engineering and Science Technology A Partnership Between Ryerson University and St.Michael’s HospitalTorontoOntarioM5B 1T8Canada

Nanoscale ultrasound contrast agents,or nanobubbles,are being explored in preclinical applications ranging from vascular and cardiac imaging to targeted drug delivery in cancer.These sub-micron particles are approximately 10x smaller than clinically available microbubbles.This allows them to effectively traverse compromised physiological barriers and circulate for extended periods of time.While various aspects of nanobubble behavior have been previously examined,their behavior in human whole blood has not yet been explored.Accordingly,herein we examined,for the first time,the short and long-term effects of blood components on nanobubble acoustic response.We observed differences in the kinetics of backscatter from nanobubble suspensions in whole blood compared to bubbles in phosphate buffered saline(PBS),plasma,or red blood cell solutions(RBCs).Specifically,after introducing nanobubbles to fresh human whole blood,signal enhancement,or the magnitude of nonlinear ultrasound signal,gradually increased by 22.8±13.1%throughout our experiment,with peak intensity reached within 145 s.In contrast,nanobubbles in PBS had a stable signal with negligible change in intensity(􀀀1.7±3.2%)over 8 min.Under the same conditions,microbubbles made with the same lipid formulation showed a􀀀56.8±6.1%decrease in enhancement in whole blood.Subsequent confocal,fluorescent,and scanning electron microscopy analysis revealed attachment of the nanobubbles to the surface of RBCs,suggesting that direct interactions,or hitchhiking,of nanobubbles on RBCs in the presence of plasma may be a possible mechanism for the observed effects.This phenomenon could be key to extending nanobubble circulation time and has broad implications in drug delivery,where RBC interaction with nanoparticles could be exploited to improve delivery efficiency.

关键词： Nanobubbles Ultrasound contrast agents red blood cells Hitchhiking Ultrasound Drug delivery