Introduction
When about 50 years ago Wolf1 noticed that some tiny and highly abundant objects present in human blood plasma contribute to clotting events, it seemed that there is something beyond clotting factors that may support platelets / thrombocytes in their aggregation and formation of a thrombus. This coagulant particulate material released from platelets (referred to as “platelet dust”) was produced in considerably larger amounts than required for thrombin generation. It was detected not only in plasma but also in serum, and its presence seemed to be associated with the platelet-like activity of the serum.1 The platelet activation leads to secretion of granules containing the procoagulant material and proteins involved in cytoskeletal arrangement, synaptic transportation, and secretion from their internal space.2 The processes of the formation and secretion of platelet “particles” are analogous to the processes occurring in other cells, including endothelial cells. They result in the formation of 2 types of membrane vesicles: bigger ones shed from the surface and named microvesicles (ectosomes) of 100 nm to 1 µm in diameter, and exosomes, measuring 40 nm to 100 nm in diameter. The latter are similar in size to the internal vesicles in multivesicular bodies (MVBs) and α-granules,3,4 and can be compared to endothelial vesiculation (Figure 1). Currently, the topic of extracellular vesicles (EVs) and their involvement in disease promotion and progression is gaining important insight in diagnostics and treatment.5 In this review, we focus on specific EV characteristics, still not intensively investigated, which contribute to the biological activity of EVs, that is, composition of the EV surface called a corona, and its charge.
The biological activity of EVs depends on their cargo and their biological availability and biostability. The estimated blood plasma concentration of EVs in healthy individuals is between 108 and 1014 EVs/ml. Such discrepancies, more than 6 orders of magnitude, depend mainly on the isolation protocol and possible contamination by other colloidal particles present in the plasma, such as lipoproteins and large protein aggregates.6 In total platelet poor plasma, the average platelet microvesicle (PMV) content has been estimated at 109 to 1010 EVs/ml in patients on antiplatelet drugs and healthy individuals. PMV abundance depends mainly on the plasma purity and preanalytical handling.7-11 Assuming that one-third of the plasma EVs is of platelet origin, the number of the plasma EVs can be approximately 1010 EVs/ml.6 The average plasma residence time of intravenously delivered EVs ranges from 30 to 80 minutes and it is mainly regulated by the phagocyting activity of the mononuclear phagocyte system and by continuous turnover of EVs secreted by cells.12,13 Phagocytosis is one of the proposed mechanisms of EV internalization.14,15 Alternatively, EVs can be internalized by target cells in a variety of endocytic pathways (eg, clathrin-dependent endocytosis16 and clathrin-independent pathways, such as macropinocytosis,14 lipid raft-mediated internalization, or caveole-mediated uptake).14,17-19
What do extracellular vesicles contain?
The term EVs is used to refer to all membrane vesicles constituting a population of very diverse vesicular structures of different size and molecular content3,4,20 (Figure 2). Their molecular cargo has been intensively investigated and gathered in the biggest and manually curated compendia of molecular data for protein, lipid, and RNA, known as Vesiclepedia or ExoCarta21-23 or extracellular vesicle–associated DNA database (EV-ADD).24 Both populations of EVs (ectosomes and exosomes) contain or carry specific proteins, which can be considered potential biomarkers.25-27 For the endothelial cells, the most pathognomonic proteins are urokinase plasminogen activator surface receptor (uPAR),28,29 von Willebrand factor (vWF),30 heat shock proteins,31 and metalloproteinases.32,33
Another molecular hallmark of EVs is microRNA (miRNA), specific, noncoding short RNA molecules.34,35 For years, EV miRNAs have been considered as a way of cell-to-cell communication. Recently, stoichiometric studies of miRNAs and exosomes have showed that most individual exosomes do not carry biologically significant amounts of miRNAs and they are unlikely to function as vehicles for miRNA-based communication.36,37 Currently, EVs are rather considered a system for removing waste from the cells, which seems to be a good alternative to traditional laboratory biomarkers,38-40 or as vehicles for drug delivery systems for further clinical use.41,42
Molecular composition of EVs differs markedly also in terms of carbohydrates (glycans) attached to the surface proteins and forming a hydrophobic sugar overcoat (Figure 3). The presence and structure of glycans play a crucial role in cellular life and functioning of glycoproteins, for example, in cell-cell recognition, pharmacokinetics, physical stability, and immunogenicity. Glycans attached to proteins exert various important biological functions, such as: 1) targeting recognition, 2) modulating protein activity, or 3) stabilizing protein folding. Changes in the glycoproteome probably contribute to the age-related functional decline of the cardiovascular system as well as the heart and the aorta performance.43,44 Glycans covering the surface of EVs form a corona that has been recognized as a crucial mediator of EV functions.45,46 Glycomic profile of melanoma-derived ectosomes showed important correlation with melanoma malignancy.47 Differences in the surface glycosylation pattern, particularly in N-acetylglucosamine, mannose, and fucose-binding lectins result in facilitated EV-cell interactions and functional activation of endothelial cells.48 Glycans are key players in the regulation of EV uptake, through charge-based effects or direct glycan recognition by targeting receptors.49
The importance of protein glycosylation for the biotechnology industry is highlighted by the fact that approximately 70% of therapeutic proteins, approved or in (pre-)clinical studies, are glycoproteins.
New feature of extracellular vesicles: a corona charge
Modification of surface N-glycans increases EV uptake and reduces EV charge, expressed as the zeta potential (ZP), from negative toward neutral. The other contributors to the surface charge are phospholipids, the most common of which is a negatively charged phosphate group (PO43−). This group, when covalently bound to the lipid glycerol moiety of a 2-chain fatty acid, forms the main group of charged fatty acids. Formation of EVs is very closely associated with the exposure of a membrane phospholipid phosphatidylserine (PS). Under normal conditions, PS is usually present in the inner membrane leaflet, but during EV secretion PS is transferred to the outer membrane leaflet.51 PS is composed of a negatively charged phosphate group attached to the serine at the hydroxyl end.
ZP is the measure that indicates the accumulation of negatively charged phospholipids in the inner membrane leaflet. Such accumulation of ions generates the ZP with an effective range of approximately 1 nm (Figure 4). Positively charged ions are attracted to the anionic surface, which is especially noticeable at the inner leaflet of the plasma membrane.
ZP is the electrostatic potential present at the boundary between the diffuse layer and the compact layer (also known as the raft layer) of a colloid system, in the case of EVs, it is an EV suspension in a body fluid. This potential is related to the surface charge of the EVs and is used as an indicator of their stability and ability to form aggregates. The more negative the ZP, the weaker the forces attaching EVs together, and the stronger the EV affinity to positive or less negative surfaces. The cell surface charge varies between positive and negative electric state, and it depends on the balance between negatively and positively charged molecules. In physiological conditions, the surface charge of endothelial cells is less negative,52 and it changes toward higher values in pathological conditions or during cancerogenesis.53-55
ZP as an indicator of colloidal stability of dispersed particles is influenced by the charge of a colloid surface. ZP is one of the useful measures to characterize colloidal stability, including EV aggregation and their electrophoretic mobility.56 Biological membranes (including EVs) bear negatively charged glycoproteins and glycolipids forming a complex corona with the surrounding medium and regulating biological properties, such as adhesiveness and internalization.57,58 The charge of the EV corona depends on different factors that control the interactions between the particle surface and the medium, such as pH and ionic strength of the medium56 (Figure 4).
As shown in Table 1, values of EV ZP differ surprisingly, ranging from –20 to –10 mV for EVs isolated from macrophages, astrocytes, and neurons57 or from –40 to –30 mV for EVs isolated from cancer cells or erythrocytes.54,58 These variations in ZP have not been explored yet, and may result from diverse separation methods, contamination of EVs with different colloids including lipoproteins, as well as external conditions, such as low pH, high ionic strength, or valency of surface cations.29,30
Origin | EV isolation methodology | EV zeta potential |
---|---|---|
Adipose-derived stem cells | 300 g, 10 min 2000 g, 10 min 10 000 g, 30 min 100 000 g, 70 min | −10.8 ± 0.65 mV59 |
Human serum | TEIR ExoQuick miRCURY Ultracentrifugation (20 000 g, 30 min; 110 000 g, 70 min) | From –9.80 mV to –21.1 mV60 |
Bone-marrow MSCs | 300 g, 5 min 16 500 g, 40 min Filtration, 0.2 µm 120 000 g, 70 min | −30 ± 1.13 mV61 |
HLSC culture media Human serum and saliva | 3000 g, 20 min Filtration, 0.22 µm 3000 g, 20 min 10 000 g, 20 min 100 000 g, 60 min | HLSC culture medium –13.80 mV Human serum –7.825 mV Saliva –8.54 mV62 |
Glioblastoma cells | Exospin exosome purification kit | Empty exosome −22.18 ± 8.73 mV Incubation at 37 °C −18.22 ± 1.23 mV Sonication, −22.46 ± 0.63 mV63 |
Colon cancer HCT116 cell line and ASCs | 3000 g, 30 min 13 000 g, 70 min Sample concentration using an Amicon Ultra-15 Centrifugal Filter Devices (100 kDa, Millipore, Merck KGaA, Darmstadt, Germany) 120 000 g, 70 min Filtration, 0.22 µm | TEx –9.20 ± 0.41 mV AEx –7.22 ± 0.60 mV64 |
Lung cancer cells | 750 g, 15 min 2000 g, 20 min Filtration, 0.45 µm 10 000 g, 45 min Filtration, 0.22 µm 100 000 g, 90 min | −14.4 ±3.3 mV at RT for exosomes diluted in PBS with trehalose −11.8 ±1.5 mV at RT for exosomes diluted in PBS with DMSO65 |
Human NB cell lines HTLA-230, IMR-32, SH-SY5Y, and GI-LI-N | 300 g, 10 min 10 000 g, 30 min Filtration, 0.22 µm Filtration, 0.1 µm 100 000 g, 75 min Washing in PBS 2 × ultracentrifugation 100 000 g, 75 min | HTLA-230 −12.1 ±0.17 mV IMR-32 −14.8 ±1.55 mV SH-SY5Y −13.2 ±1.1 mV GI-LI-N −12 ±0.15 mV66 |
Human umbilical cord–derived mesenchymal stromal cells | 3200 g, 30 min Filtration, 0.2 µm Sample concentration using an Amicon Ultra-15 filter unit with Ultracel-100 membrane (MWCO = 100 kDa, Merck Millipore, Cat. No. UFC910024) SEC column | From −7.73 ±3.76 mV to −12.4 ±2.5 mV67 |
Human urine–derived stem cells | Centrifugation using an Amicon Ultra15 Centrifugal Filter Tube (10 kDa; Millipore) ExoQuick-TC Solution (System Biosciences, Palo Alto, California, United States) 1500 g, 30 min | −2.02 ±0.03 mV68 |
Red blood cells | 1500 g, 10 min 3000 g, 15 min 25 000 g, 60 min ~200 g, 120 min | −36.4 ±7.8 mV69 |
Raw bovine milk | 13 000 g, 30 min 90 000 g, 60 min 180 000 g, 120 min Filtration, 0.2 µm | Exo-PAC −28.28 ±1.8 mV Exo-5-FU −27 ±1.6 mV; plain exosomes −23 ±1.2 mV70 |
Human serum | Total exosome isolation from serum (Invitrogen by Thermo Fisher Scientific, Vilnius, Lithuania) | Serum small EVs (sEVs) from RB individuals −11.04 ±0.4 mV Serum small EVs (sEVs) from non-RB individuals –12.72 ±1.7 mV71 |
Murine cardiac fibroblasts (CF) and CF-derived iPS | 3000 g, 10 min Filtration, 0.2 µm precipitation overnight in PEG buffer at 4 ˚C 1500 g, 30 min | CF exosomes −14.22mV iPS exosomes −15.44 mV72 |
Human immortalized microvascular endothelial cell line (TIME) | 2000 g, 30 min 18 000 g, 30 min 150 000 g, 90 min | Ectosomes –9.3 ±0.7 mV Exosomes –11.35 ±1.9 mV22 |
Abbreviations: AEx, adipose stem cells–derived exosomes; ASCs, adipose stem cells; DMSO, dimethyl sulfoxide; Exo-5-FU, 5-fluorouracil-loaded exosomes; Exo-PAC; paclitaxel-loaded exosomes; HLSC, adult human liver stem cells; iPS, induced pluripotent stem cells; MLC, mixed lymphocyte culture; MWCO, molecular weight cutoff; NB, neuroblastoma; PBS, phosphate-buffered saline; RB, retinoblastoma; SEC, size exclusion chromatograhy; TEIR, Total Exosome Isolation Reagent for serum; TEx, tumor cell-derived exosomes |
Concluding remarks
EVs are very attractive research objects as potential biomarkers of various pathologies due to their specific cargo and abundance in the cellular environment. As they have different cellular origin, they also differ in their metabolic competencies, and carry a variety of bioactive molecules, such as miRNAs, proteins, and lipids. However, their biodistribution and availability are also important and still underestimated features. For their further use in clinical practice as potential drug carriers, their biodistribution and uptake by target cells should be considered. To better understand the mechanisms of cell internalization, most attention should be given to the EV surface properties related to the EV corona and surface charge, characterized by glycosylation and ZP.
Ewa Ł. Stępień, PhD, Marian Smoluchowski Institute of Physics, Jagiellonian University, ul. Łojasiewicza 11, 30-348 Kraków, Poland, phone: +48 12 664 47 62, email: e.stepien@uj.edu.pl
March 22, 2023.
March 23, 2023.
April 19, 2023.
The authors thank Dr. Karol Kubat (Jagiellonian University) who assisted in the preparation of Figure 3.
None declared.
Stępień EŁ, Durak-Kozica M, Moskal P. Extracellular vesicles in vascular pathophysiology: beyond their molecular content. Pol Arch Intern Med. 2023; 133: 16483. doi:10.20452/pamw.16483
- 1.
- Wolf P. The nature and significance of platelet products in human plasma. Br J Haematol. 1967; 13: 269-288.Crossref
- 2.
- Kasprzyk J, Stępień E, Piekoszewski W. Application of nano-LC-MALDI-TOF/TOF-MS for proteomic analysis of microvesicles. Clin Biochem. 2017; 50: 241-243.Crossref
- 3.
- Heijnen HF, Schiel AE, Fijnheer R, et al. Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood. 1999; 94: 3791-3799.Crossref
- 4.
- Cocucci E, Meldolesi J. Ectosomes and exosomes: shedding the confusion between extracellular vesicles. Trends Cell Biol. 2015; 25: 364-372.Crossref
- 5.
- Stępień E, Rząca C, Moskal P. Novel biomarker and drug delivery systems for theranostics - extracellular vesicles. Bio-Algorithms and Med-Syst. 2021; 17: 301-309.Crossref
- 6.
- Johnsen KB, Gudbergsson JM, Andresen TL, Simonsen JB. What is the blood concentration of extracellular vesicles? Implications for the use of extracellular vesicles as blood-borne biomarkers of cancer. Biochim Biophys Acta Rev Cancer. 2019; 1871: 109-116.Crossref
- 7.
- Chandler WL. Microparticle counts in platelet-rich and platelet-free plasma, effect of centrifugation and sample-processing protocols. Blood Coagul Fibrinolysis. 2013; 24: 125-132.Crossref
- 8.
- Stępień E, Gruszczyński K, Kapusta P, et al. Plasma centrifugation does not influence thrombin-antithrombin and plasmin-antiplasmin levels but determines platelet microparticles count. Biochem Med (Zagreb). 2015; 25: 222-229.Crossref
- 9.
- Cointe S, Judicone C, Robert S, et al. Standardization of microparticle enumeration across different flow cytometry platforms: results of a multicenter collaborative workshop. J Thromb Haemost. 2017; 15: 187-193.Crossref
- 10.
- Lacroix R, Judicone C, Mooberry M, et al; The ISTH SSC Workshop. Standardization of pre-analytical variables in plasma microparticle determination: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop. J Thromb Haemost. 2013 Apr 2. [Epub ahead of print]Crossref
- 11.
- Buntsma NC, Gąsecka A, Roos YBWEM, et al. EDTA stabilizes the concentration of platelet-derived extracellular vesicles during blood collection and handling. Platelets. 2022; 33: 764-771.Crossref
- 12.
- Charoenviriyakul C, Takahashi Y, Nishikawa M, Takakura Y. Preservation of exosomes at room temperature using lyophilization. Int J Pharmaceutics. 2018; 553: 1-7.Crossref
- 13.
- Lai CP, Mardini O, Ericsson M, et al. Dynamic biodistribution of extracellular vesicles in vivo using a multimodal imaging reporter. ACS Nano. 2014; 8: 483-494.Crossref
- 14.
- Doherty GJ, McMahon HT. Mechanisms of endocytosis. Ann Rev Biochem. 2009; 78: 857-902.Crossref
- 15.
- Swanson JA. Shaping cups into phagosomes and macropinosomes. Nat Rev Mol Cell Biol. 2008; 9: 639-649.Crossref
- 16.
- Wang LH, Rothberg KG, Anderson RG. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol. 1993; 123: 1107-1117.Crossref
- 17.
- Nanbo A, Kawanishi E, Yoshida R, Yoshiyama H. Exosomes derived from Epstein-Barr virus-infected cells are internalized via caveola-dependent endocytosis and promote phenotypic modulation in target cells. J Virol. 2013; 87: 10334-10347.Crossref
- 18.
- Wąchalska M, Rychłowski M, Grabowska K, et al. Palmitoylated mNeonGreen protein as a tool for visualization and uptake studies of extracellular vesicles. Membranes (Basel). 2020; 10: 373.Crossref
- 19.
- El-Sayed A, Harashima H. Endocytosis of gene delivery vectors: from clathrin-dependent to lipid raft-mediated endocytosis. Mol Ther. 2013; 21: 1118-1130.Crossref
- 20.
- Durak-Kozica M, Wróbel A, Platt M, Stępień EŁ. Comparison of qNANO results from the isolation of extracellular microvesicles with the theoretical model. Bio-Algorithms and Med-Systems. 2022; 18: 171-179.Crossref
- 21.
- Kalra H, Simpson RJ, Ji H, et al. Vesiclepedia: a compendium for extracellular vesicles with continuous community annotation. PLoS Biol. 2012; 10: e1001450.Crossref
- 22.
- Pathan M, Fonseka P, Chitti SV, et al. Vesiclepedia 2019: a compendium of RNA, proteins, lipids and metabolites in extracellular vesicles. Nucleic Acids Res. 2019; 47: D516-D519.Crossref
- 23.
- Keerthikumar S, Chisanga D, Ariyaratne D, et al. ExoCarta: a web-based compendium of exosomal cargo. J Mol Biol. 2016; 428: 688-692.Crossref
- 24.
- Tsering T, Li M, Chen Y, et al. EV-ADD, a database for EV-associated DNA in human liquid biopsy samples. J Extracell Vesicles. 2022; 11: e12270.Crossref
- 25.
- Hurwitz SN, Rider MA, Bundy JL, et al. Proteomic profiling of NCI-60 extracellular vesicles uncovers common protein cargo and cancer type-specific biomarkers. Oncotarget. 2016; 7: 86999-87015.Crossref
- 26.
- Stępień E, Kabłak-Ziembicka A, Czyż J, et al. Microparticles, not only markers but also a therapeutic target in the early stage of diabetic retinopathy and vascular aging. Expert Opin Ther Targets. 2012; 16: 677-688.Crossref
- 27.
- Tokarz A, Konkolewska M, Kuśnierz-Cabala B, et al. Retinopathy severity correlates with RANTES concentrations and CCR 5-positive microvesicles in diabetes. Folia Med Cracov. 2019; 59: 95-112.
- 28.
- Durak-Kozica M, Enguita FJ, Stępień E. Targeting uPAR in diabetic vascular pathologies. Postepy Hig Med Dosw. 2019; 73: 803-808.Crossref
- 29.
- Durak-Kozica M, Baster Z, Kubat K, Stępień E. 3D visualization of extracellular vesicle uptake by endothelial cells. Cell Mol Biol Lett. 2018; 23: 57.Crossref
- 30.
- Peterson DB, Sander T, Kaul S, et al. Comparative proteomic analysis of PAI-1 and TNF-alpha-derived endothelial microparticles. Proteomics. 2008; 8: 2430-2446.Crossref
- 31.
- Taha EA, Ono K, Eguchi T. Roles of extracellular HSPs as biomarkers in immune surveillance and immune evasion. Int J Mol Sci. 2019; 20: 4588.Crossref
- 32.
- Adnani L, Kassouf J, Meehan B, et al. Angiocrine extracellular vesicles impose mesenchymal reprogramming upon proneural glioma stem cells. Nat Commun. 2022; 13: 5494.Crossref
- 33.
- Taraboletti G, D’Ascenzo S, Borsotti P, et al. Shedding of the matrix metalloproteinases MMP-2, MMP-9, and MT1-MMP as membrane vesicle-associated components by endothelial cells. Am J Pathol. 2002; 160: 673-680.Crossref
- 34.
- Lotvall J, Valadi H. Cell to cell signalling via exosomes through esRNA. Cell Adh Migr. 2007; 1: 156-158.Crossref
- 35.
- Alexandru N, Badila E, Weiss E, et al. Vascular complications in diabetes: microparticles and microparticle associated microRNAs as active players. Biochem Biophys Res Commun. 2016; 472: 1-10.Crossref
- 36.
- Chevillet JR, Kang Q, Ruf IK, et al. Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc Natl Acad Sci U S A. 2014; 111: 14888-14893.Crossref
- 37.
- Zapała B, Kamińska A, Piwowar M, et al. miRNA signature of urine extracellular vesicles shows the involvement of inflammatory and apoptotic processes in diabetic chronic kidney disease. Pharm Res. 2023 Mar 1. [Epub ahead of print]Crossref
- 38.
- Sekuła M, Janawa G, Stankiewicz E, Stepień E. Endothelial microparticle formation in moderate concentrations of homocysteine and methionine in vitro. Cell Mol Biol Lett. 2011; 16: 69-78.Crossref
- 39.
- Stępień EŁ, Durak-Kozica M, Kamińska A, et al. Circulating ectosomes: determination of angiogenic microRNAs in type 2 diabetes. Theranostics. 2018; 8: 3874-3890.Crossref
- 40.
- Kamińska A, Roman M, Wróbel A, et al. Raman spectroscopy of urinary extracellular vesicles to stratify patients with chronic kidney disease in type 2 diabetes. Nanomedicine. 2022; 39: 102468.Crossref
- 41.
- Stępień EŁ, Rząca C, Moskal P. Radiovesicolomics - new approach in medical imaging. Front Physiol. 2022; 13: 996985.Crossref
- 42.
- Rząca C, Jankowska U, Stępień EŁ. Proteomic profiling of exosomes derived from pancreatic beta-cells cultured under hyperglycemia. Bio-Algorithms and Med-Systems. 2022; 18: 151-157.Crossref
- 43.
- Przybyło M, Stepień E, Pfitzner R, et al. Age effect on human aortic valvular glycoproteins. Arch Med Res. 2007; 38: 495-502.Crossref
- 44.
- Franzka P, Krüger L, Schurig MK, et al. Altered glycosylation in the aging heart. Front Mol Biosci. 2021; 8: 673044.Crossref
- 45.
- Tóth EÁ, Turiák L, Visnovitz T, et al. Formation of a protein corona on the surface of extracellular vesicles in blood plasma. J Extracell Vesicles. 2021; 10: e12140.Crossref
- 46.
- Buzas EI. Opportunities and challenges in studying the extracellular vesicle corona. Nat Cell Biol. 2022; 24: 1322-1325.Crossref
- 47.
- Surman M, Hoja-Łukowicz D, Szwed S, et al. Human melanoma-derived ectosomes are enriched with specific glycan epitopes. Life Sci. 2018; 207: 395-411.Crossref
- 48.
- Clos-Sansalvador M, Garcia SG, Morón-Font M, et al. N-Glycans in immortalized mesenchymal stromal cell-derived extracellular vesicles are critical for EV-cell interaction and functional activation of endothelial cells. Int J Mol Sci. 2022; 23: 9539.Crossref
- 49.
- Williams C, Pazos R, Royo F, et al. Assessing the role of surface glycans of extracellular vesicles on cellular uptake. Sci Rep. 2019; 9: 11920.Crossref
- 50.
- Williams C, Royo F, Aizpurua-Olaizola O, et al. Glycosylation of extracellular vesicles: current knowledge, tools and clinical perspectives. J Extracell Vesicles. 2018; 7: 1442985.Crossref
- 51.
- Marzec ME, Rząca C, Moskal P, Stępień EŁ. Study of the influence of hyperglycemia on the abundance of amino acids, fatty acids, and selected lipids in extracellular vesicles using TOF-SIMS. Biochem Biophys Res Commun. 2022; 622: 30-36.Crossref
- 52.
- Ghitescu L, Fixman A. Surface charge distribution on the endothelial cell of liver sinusoids. J Cell Biol. 1984; 99: 639-647.Crossref
- 53.
- Ghinea N, Simionescu N. Anionized and cationized hemeundecapeptides as probes for cell surface charge and permeability studies: differentiated labeling of endothelial plasmalemmal vesicles. J Cell Biol. 1985; 100: 606-612.Crossref
- 54.
- Vargas FF, Osorio MH, Ryan US, De Jesus, M. Surface charge of endothelial cells estimated from electrophoretic mobility. Membr Biochem. 1989; 8: 221-227.Crossref
- 55.
- Chen B, Le W, Wang Y, et al. Targeting negative surface charges of cancer cells by multifunctional nanoprobes. Theranostics. 2016; 6: 1887-1898.Crossref
- 56.
- Midekessa G, Godakumara K, Ord J, et al. Zeta potential of extracellular vesicles: toward understanding the attributes that determine colloidal stability. ACS Omega. 2020; 5: 16701-16710.Crossref
- 57.
- Haney MJ, Zhao Y, Fallon JK, et al. Extracellular vesicles as drug delivery system for treatment of neurodegenerative disorders: optimization of the cell source. Adv Nanobiomed Res. 2021; 1: 2100064.Crossref
- 58.
- Nakase I, Ueno N, Matsuzawa M, et al. Environmental pH stress influences cellular secretion and uptake of extracellular vesicles. FEBS Open Bio. 2021; 11: 753-767.Crossref
- 59.
- Li R, Li D, Wang H, et al. Exosomes from adipose-derived stem cells regulate M1/M2 macrophage phenotypic polarization to promote bone healing via miR-451a/MIF. Stem Cell Res Ther. 2022; 13: 149.Crossref
- 60.
- Helwa I, Cai J, Drewry MD, et al. A comparative study of serum exosome isolation using differential ultracentrifugation and three commercial reagents. PloS One. 2017; 12: e0170628.Crossref
- 61.
- Ashour AA, El-Kamel AH, Mehanna RA, et al. Luteolin-loaded exosomes derived from bone marrow mesenchymal stem cells: a promising therapy for liver fibrosis. Drug Delivery. 2022; 29: 3270-3280.Crossref
- 62.
- Deregibus MC, Figliolini F, D’antico S, et al. Charge-based precipitation of extracellular vesicles. Int J Mol Med. 2016; 38: 1359-1366.Crossref
- 63.
- Salarpour S, Forootanfar H, Pournamdari M, et al. Paclitaxel incorporated exosomes derived from glioblastoma cells: comparative study of two loading techniques. Daru. 2019; 27: 533-539.Crossref
- 64.
- Jing B, Gai Y, Qian R, et al. Hydrophobic insertion-based engineering of tumor cell-derived exosomes for SPECT/NIRF imaging of colon cancer. J Nanobiotechnology. 2021; 19: 1-13.Crossref
- 65.
- Ruzycka-Ayoush M, Nowicka AM, Kowalczyk A, et al. 2023. Exosomes derived from lung cancer cells: Isolation, characterization, and stability studies. Eur J Pharm Sci. 2023; 181: 106369.Crossref
- 66.
- Marimpietri D, Petretto A, Raffaghello L, et al. Proteome profiling of neuroblastoma-derived exosomes reveal the expression of proteins potentially involved in tumor progression. PloS One. 2013; 8: e75054.Crossref
- 67.
- Wright A, Snyder OL, Christenson LK, et al. Effect of pre-processing storage condition of cell culture-conditioned medium on extracellular vesicles derived from human umbilical cord-derived mesenchymal stromal cells. Int J Mol Sci. 2022; 23: 7716.Crossref
- 68.
- Li H, Hu Y, Zeng M, et al. exosomes from human urine-derived stem cells encapsulated into PLGA nanoparticles for therapy in mice with particulate polyethylene-induced osteolysis. Front Med. 2021; 8: 781449.Crossref
- 69.
- Nguyen DB, Tran HT, Kaestner L, Bernhardt I. The relation between extracellular vesicles released from red blood cells, their cargo, and the clearance by macrophages. Front Physiol. 2022; 13: 783260.Crossref
- 70.
- Kumar DN, Chaudhuri A, Dehari D, et al. Combination therapy comprising paclitaxel and 5-fluorouracil by using folic acid functionalized bovine milk exosomes improves the therapeutic efficacy against breast cancer. Life. 2022; 12: 1143.Crossref
- 71.
- Manukonda R, Yenuganti VR, Nagar N, et al. Comprehensive analysis of serum small extracellular vesicles-derived coding and non-coding RNAs from retinoblastoma patients for identifying regulatory interactions. Cancers. 2022; 14: 4179.Crossref
- 72.
- Wang Y, Zhang L, Li Y, et al. Exosomes / microvesicles from induced pluripotent stem cells deliver cardioprotective miRNAs and prevent cardiomyocyte apoptosis in the ischemic myocardium. Int J Cardiol. 2015; 192: 61-69.Crossref