Nano drug-delivery systems (DDS) may significantly improve efficiency and reduce toxicity of loaded drugs, but a few nano-DDS are highly successful in clinical use. Unprotected nanoparticles in blood flow are often quickly cleared, which could limit their circulation time and drug delivery efficiency. Elongating their blood circulation time may improve their delivery efficiency or grant them new therapeutic possibilities. Erythrocytes are abundant endogenous cells in blood and are continuously renewed, with a long life span of 100–120 days. Hence, loading nanoparticles on the surface of erythrocytes to protect the nanoparticles could be highly effective for enhancing their in vivo circulation time. One of the key questions here is how to properly attach nanoparticles on erythrocytes for different purposes and different types of nanoparticles to achieve ideal results. In this review, we describe various methods to attach nanoparticles and drugs to the erythrocyte surface, and discuss the key factors that influence the stability and circulation properties of the erythrocytes-based delivery system in vivo. These data show that using erythrocytes as a host for nanoparticles possesses great potential for further development.
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Hu CMJ, Fang RH, Zhang LF. Erythrocyte-inspired delivery systems. Adv Healthc Mater. 2012;1:537–47.
Mohandas N, Gallagher PG. Red cell membrane: past, present, and future. Blood. 2008;112:3939–48.
Sheng Y, Liu CS, Yuan Y, Tao XY, Yang F, Shan XQ, et al. Long-circulating polymeric nanoparticles bearing a combinatorial coating of PEG and water-soluble chitosan. Biomaterials. 2009;30:2340–8.
Chambers E, Mitragotri S. Prolonged circulation of large polymeric nanoparticles by non-covalent adsorption on erythrocytes. J Control Release. 2004;100:111–9.
Yoo JW, Chambers E, Mitragotri S. Factors that control the circulation time of nanoparticles in blood: challenges, solutions and future prospects. Curr Pharm Des. 2010;16:2298–307.
Schipper ML, Iyer G, Koh AL, Cheng Z, Ebenstein Y, Aharoni A, et al. Particle size, surface coating, and PEGylation influence the biodistribution of quantum dots in living mice. Small. 2009;5:126–34.
Hoshyar N, Gray S, Han HB, Bao G. The effect of nanoparticle size on in vivo pharmacokinetics and cellular interaction. Nanomedicine. 2016;11:673–92.
Albanese A, Tang PS, Chan WC. The effect of nanoparticle size, shape, and surface chemistry on biological systems. Annu Rev Biomed Eng. 2012;14:1–16.
Pan D, Vargas-Morales O, Zern B, Anselmo AC, Gupta V, Zakrewsky M, et al. The effect of polymeric nanoparticles on biocompatibility of carrier red blood cells. PLoS One. 2016;11:e152074.
Toy R, Hayden E, Shoup C, Baskaran H, Karathanasis E. The effects of particle size, density and shape on margination of nanoparticles in microcirculation. Nanotechnology. 2011;22:115101.
Ta HT, Truong NP, Whittaker AK, Davis TP, Peter K. The effects of particle size, shape, density and flow characteristics on particle margination to vascular walls in cardiovascular diseases. Expert Opin Drug Deliv. 2018;15:33–45.
Magnani M, Pierigè F, Rossi L. Erythrocytes as a novel delivery vehicle for biologics: from enzymes to nucleic acid-based therapeutics. Ther Deliv. 2012;3:405–14.
Brahler M, Georgieva R, Buske N, Muller A, Muller S, Pinkernelle J, et al. Magnetite-loaded carrier erythrocytes as contrast agents for magnetic resonance imaging. Nano Lett. 2006;6:2505–9.
Muzykantov VR. Drug delivery by red blood cells: vascular carriers designed by mother nature. Expert Opin Drug Deliv. 2010;7:403–27.
Chambers E, Mitragotri S. Long circulating nanoparticles via adhesion on red blood cells: mechanism and extended circulation. Exp Biol Med. 2007;232:958.
Sahoo K, Koralege RSH, Flynn N, Koteeswaran S, Clark P, Hartson S, et al. Nanoparticle attachment to erythrocyte via the glycophorin A targeted ERY1 ligand enhances binding without impacting cellular function. Pharmacol Res. 2016;33:1191–203.
Fu Y, Liu W, Wang LY, Zhu BY, Qu MK, Yang LQ, et al. Erythrocyte-membrane-camouflaged nanoplatform for intravenous glucose-responsive insulin delivery. Adv Funct Mater. 2018;28:1802250.
Villa CH, Anselmo AC, Mitragotri S, Muzykantov V. Red blood cells: supercarriers for drugs, biologicals, and nanoparticles and inspiration for advanced delivery systems. Adv Drug Deliv Rev. 2016;106:88–103.
Jensen FB. The dual roles of red blood cells in tissue oxygen delivery: oxygen carriers and regulators of local blood flow. J Exp Biol. 2009;212:3387–93.
Hamidi M, Zarrin A, Foroozesh M, Mohammadi-Samani S. Applications of carrier erythrocytes in delivery of biopharmaceuticals. J Control Release. 2007;118:145–60.
Rossi L, Pierige F, Antonelli A, Bigini N, Gabucci C, Peiretti E, et al. Engineering erythrocytes for the modulation of drugs’ and contrasting agents’ pharmacokinetics and biodistribution. Adv Drug Deliv Rev. 2016;106:73–87.
He HN, Ye JX, Wang YS, Liu Q, Chung HS, Kwon YM, et al. Cell-penetrating peptides meditated encapsulation of protein therapeutics into intact red blood cells and its application. J Control Release. 2014;176:123–32.
Villa CH, Pan DC, Zaitsev S, Cines DB, Siegel DL, Muzykantov VR. Delivery of drugs bound to erythrocytes: new avenues for an old intravascular carrier. Ther Deliv. 2015;6:795–826.
Bird J, Best R, Lewis DA. The encapsulation of insulin in erythrocytes. J Pharm Pharmacol. 1983;35:246–7.
Harisa G, Ibrahim MF, Alanazi FK. Characterization of human erythrocytes as potential carrier for pravastatin: an in vitro study. Int J Med Sci. 2011;8:222–30.
Beutler E, Dale GL, Guinto E, Kuhl W. Enzyme replacement therapy in Gaucher’s disease preliminary clinical trial of a new enzyme preparation. Proc Natl Acad Sci USA. 1977;74:4620–3.
Baysal SH, Uslan AH, Pala HH, Tuncoku O. Encapsulation of PEG-Urease/PEG-AlaDH within sheep erythrocytes and determination of the system’s activity in lowering blood levels of urea in animal models. Artif Cells Blood Substit Immobil Biotechnol. 2007;35:391–403.
Millan CG, Castaneda AZ, Marinero MLS, Lanao JM. Factors associated with the performance of carrier erythrocytes obtained by hypotonic dialysis. Blood Cells Mol Dis. 2004;33:132–40.
Bax BE, Bain MD, Fairbanks LD, Webster ADB, Chalmers RA. In vitro and in vivo studies with human carrier erythrocytes loaded with polyethylene glycol-conjugated and native adenosine deaminase. Br J Haematol. 2000;109:549–54.
Banz A, Cremel M, Rembert A, Godfrin Y. In situ targeting of dendritic cells by antigen-loaded red blood cells: A novel approach to cancer immunotherapy. Vaccine. 2010;28:2965–72.
Wang GP, Guan YS, Jin XR, Jiang SS, Lu ZJ, Wu Y, et al. Development of novel 5-fluorouracil carrier erythrocyte with pharmacokinetics and potent antitumor activity in mice bearing malignant ascites. J Gastroenterol Hepatol. 2010;25:985–90.
Gallagher PG, Chang SH, Rettig MP, Neely JE, Hillery CA, Smith BD, et al. Altered erythrocyte endothelial adherence and membrane phospholipid asymmetry in hereditary hydrocytosis. Blood. 2003;101:4625–7.
Podsiedlik M, Markowicz-Piasecka M, Sikora J. Erythrocytes as model cells for biocompatibility assessment, cytotoxicity screening of xenobiotics and drug delivery. Chem Biol Interact. 2020;332:109305.
Villa CH, Cines DB, Siegel DL, Muzykantov V. Erythrocytes as carriers for drug delivery in blood transfusion and beyond. Transfus Med Rev. 2017;31:26–35.
Gao XL, Yue TT, Tian FL, Liu ZP, Zhang XR. Erythrocyte membrane skeleton inhibits nanoparticle endocytosis. AIP Adv. 2017;7:65303.
Zhao ZM, Ukidve A, Gao YS, Kim J, Mitragotri S. Erythrocyte leveraged chemotherapy (ELeCt): nanoparticle assembly on erythrocyte surface to combat lung metastasis. Sci Adv. 2019;5:eaax9250.
Pishesha N, Bilate AM, Wibowo MC, Huang NJ, Li ZY, Dhesycka R, et al. Engineered erythrocytes covalently linked to antigenic peptides can protect against autoimmune disease. Proc Natl Acad Sci USA. 2017;114:3157–62.
Glassman PM, Villa CH, Ukidve A, Zhao ZM, Smith P, Mitragotri S, et al. Vascular drug delivery using carrier red blood cells: focus on RBC surface loading and pharmacokinetics. Pharmaceutics. 2020;12:440.
Han X, Wang C, Liu Z. Red blood cells as smart delivery systems. Bioconjug Chem. 2018;29:852–60.
Yan JJ, Yu JC, Wang C, Gu Z. Red blood cells for drug delivery. Small Methods. 2017;1:1700270.
Zaitsev S, Danielyan K, Murciano JC, Ganguly K, Krasik T, Taylor RP, et al. Human complement receptor type 1–directed loading of tissue plasminogen activator on circulating erythrocytes for prophylactic fibrinolysis. Blood. 2006;108:1895–902.
Zaitsev S, Spitzer D, Murciano JC, Ding BS, Tliba S, Kowalska MA, et al. Targeting of a mutant plasminogen activator to circulating red blood cells for prophylactic fibrinolysis. J Pharmacol Exp Ther. 2010;332:1022–31.
Domenech C, Thomas X, Chabaud S, Baruchel A, Gueyffier F, Mazingue F, et al. l-asparaginase loaded red blood cells in refractory or relapsing acute lymphoblastic leukaemia in children and adults: results of the GRASPALL 2005-01 randomized trial. Br J Haematol. 2011;153:58–65.
Anselmo AC, Kumar S, Gupta V, Pearce AM, Ragusa A, Muzykantov V, et al. Exploiting shape, cellular-hitchhiking and antibodies to target nanoparticles to lung endothelium: Synergy between physical, chemical and biological approaches. Biomaterials. 2015;68:1–8.
Brenner JS, Pan DC, Myerson JW, Marcos-Contreras OA, Villa CH, Patel P, et al. Red blood cell-hitchhiking boosts delivery of nanocarriers to chosen organs by orders of magnitude. Nat Commun. 2018;9:2684.
Red Blood Cells Harnessed as Nanoparticle Carriers for Vaccines. Genetic Engineering and Biotechnology News. 2020 July 14. https://www.genengnews.com/news/red-blood-cells-harnessed-as-nanoparticle-carriers-for-vaccines/
Ukidve A, Zhao ZM, Fehnel A, Krishnan V, Pan DIC, Gao YS, et al. Erythrocyte-driven immunization via biomimicry of their natural antigen-presenting function. Proc Natl Acad Sci USA. 2020;117:17727–36.
Anselmo AC, Gupta V, Zern BJ, Pan D, Zakrewsky M, Muzykantov V, et al. Delivering nanoparticles to lungs while avoiding liver and spleen through adsorption on red blood cells. ACS Nano. 2013;7:11129–37.
Zelepukin IV, Yaremenko AV, Shipunova VO, Babenyshev AV, Balalaeva IV, Nikitin PI, et al. Nanoparticle-based drug delivery via RBC-hitchhiking for the inhibition of lung metastases growth. Nanoscale. 2019;11:1636–46.
Scott MD, Murad KL, Koumpouras F, Talbot M, Eaton JW, et al. Chemical camouflage of antigenic determinants: Stealth erythrocytes. Proc Natl Acad Sci USA. 1997;14:7566–71.
Park J, Andrade B, Seo Y, Kim MJ, Zimmerman SC, Kong H. Engineering the surface of therapeutic “living”cells. Chem Rev. 2018;118:1664–90.
Pan DC, Myerson JW, Brenner JS, Patel PN, Anselmo AC, Mitragotri S, et al. Nanoparticle properties modulate their attachment and effect on carrier red blood cells. Sci Rep. 2018;8:1615.
Setyawati MI, Tay CY, Docter D, Stauber RH, Leong DT. Understanding and exploiting nanoparticles’ intimacy with the blood vessel and blood. Chem Soc Rev. 2015;44:8174–99.
Barshtein G, Livshits L, Shvartsman LD, Shlomai NO, Arbell D, Yedgar S. Polystyrene nanoparticles activate erythrocyte aggregation and adhesion to endothelial cells. Cell Biochem Biophys. 2016;74:19–27.
Charoenphol P, Onyskiw PJ, Carrasco-Teja M, Eniola-Adefeso O. Particle-cell dynamics in human blood flow: Implications for vascular-targeted drug delivery. J Biomech. 2012;45:2822–8.
Down LA, Papavassiliou DV, O’Rear EA. Significance of extensional stresses to red blood cell lysis in a shearing flow. Ann Biomed Eng. 2011;39:1632–42.
Chinol M, Casalini P, Maggiolo M, Canevari S, Omodeo ES, Caliceti P, et al. Biochemical modifications of avidin improve pharmacokinetics and biodistribution, and reduce immunogenicity. Br J Cancer. 1998;78:189–97.
Stephan MT, Irvine DJ. Enhancing cell therapies from the outside in: Cell surface engineering using synthetic nanomaterials. Nano Today. 2011;6:309–25.
Anselmo AC, Mitragotri S. Cell-mediated delivery of nanoparticles: Taking advantage of circulatory cells to target nanoparticles. J Control Release. 2014;190:531–41.
Anselmo AC, Kumar S, Gupta V, Pearce AM, Ragusa A, Muzykantov V, et al. Exploiting shape, cellular-hitchhiking and antibodies to target nanoparticles to lung endothelium: synergy between physical, chemical and biological approaches. Biomaterials. 2015;68:1–8.
He CB, Hu YP, Yin LC, Tang C, Yin CH. Effects of particle size and surface charge on cellular uptake and biodistribution of polymeric nanoparticles. Biomaterials. 2010;31:3657–66.
Foged C, Brodin B, Frokjaer S, Sundblad A. Particle size and surface charge affect particle uptake by human dendritic cells in an in vitro model. Int J Pharm. 2005;298:315–22.
Kulkarni SA, Feng SS. Effects of particle size and surface modification on cellular uptake and biodistribution of polymeric nanoparticles for drug delivery. Pharmacol Res. 2013;30:2512–22.
Aggarwal P, Hall JB, McLeland CB, Dobrovolskaia MA, McNeil SE. Nanoparticle interaction with plasma proteins as it relates to particle biodistribution, biocompatibility and therapeutic efficacy. Adv Drug Deliv Rev. 2009;61:428–37.
Xiao K, Li YP, Luo JT, Lee JS, Xiao WW, Gonik AM, et al. The effect of surface charge on in vivo biodistribution of PEG-oligocholic acid based micellar nanoparticles. Biomaterials. 2011;32:3435–46.
Benne N, van Duijn J, Kuiper J, Jiskoot W, Slutter B. Orchestrating immune responses: How size, shape and rigidity affect the immunogenicity of particulate vaccines. J Control Release. 2016;234:124–34.
Nowak M, Brown TD, Graham A, Helgeson ME, Mitragotri S. Size, shape, and flexibility influence nanoparticle transport across brain endothelium under flow. Bioeng Transl Med. 2020;5:e10153.
Kolhar P, Anselmo AC, Gupta V, Pant K, Prabhakarpandian B, Ruoslahti E, et al. Using shape effects to target antibody-coated nanoparticles to lung and brain endothelium. Proc Natl Acad Sci USA. 2013;110:10753–8.
Doshi N, Prabhakarpandian B, Rea-Ramsey A, Pant K, Sundaram S, Mitragotri S. Flow and adhesion of drug carriers in blood vessels depend on their shape: a study using model synthetic microvascular networks. Release. J Control Release. 2010;146:196–200.
Lee S, Ferrari M, Decuzzi P. Shaping nano-/micro-particles for enhanced vascular interaction in laminar flows. Nanotechnology. 2009;20:495101.
Shuvaev VV, Ilies MA, Simone E, Zaitsev S, Kim Y, Cai SS, et al. Endothelial targeting of antibody-decorated polymeric filomicelles. ACS Nano. 2011;5:6991–9.
Geng Y, Dalhaimer P, Cai SS, Tsai R, Tewari M, Minko T, et al. Shape effects of filaments versus spherical particles in flow and drug delivery. Nat Nanotechnol. 2007;2:249–55.
Yoo J, Mitragotri S. Polymer particles that switch shape in response to a stimulus. Proc Natl Acad Sci USA. 2010;107:11205–10.
Champion JA, Mitragotri S. Shape induced inhibition of phagocytosis of polymer particles. Pharmacol Res. 2009;26:244–9.
Gutierrez M, Ojeda LS, Eniola-Adefeso O. Vascular-targeted particle binding efficacy in the presence of rigid red blood cells: implications for performance in diseased blood. Biomicrofluidics. 2018;12:42217.
Anselmo AC, Zhang M, Kumar S, Vogus DR, Menegatti S, Helgeson ME, et al. Elasticity of nanoparticles influences their blood circulation, phagocytosis, endocytosis, and targeting. ACS Nano. 2015;9:3169–77.
Hui Y, Yi X, Wibowo D, Yang GZ, Middelberg APJ, Gao HJ, et al. Nanoparticle elasticity regulates phagocytosis and cancer cell uptake. Sci Adv. 2020;6:eaaz4316.
Guo P, Liu D, Subramanyam K, Wang BR, Yang J, Huang J, et al. Nanoparticle elasticity directs tumor uptake. Nat Commun. 2018;9:130.
Champion JA, Mitragotri S. Role of target geometry in phagocytosis. Proc Natl Acad Sci USA. 2006;103:4930–4.
Anselmo AC, Mitragotri S. Impact of particle elasticity on particle-based drug delivery systems. Adv Drug Deliv Rev. 2017;108:51–67.
Herant M. Mechanics of neutrophil phagocytosis: behavior of the cortical tension. J Cell Sci. 2005;118:1789–97.
Garapaty A, Champion JA. Tunable particles alter macrophage uptake based on combinatorial effects of physical properties. Bioeng Transl Med. 2017;2:92–101.
Carboni E, Tschudi K, Nam J, Lu XL, Ma AWK. Particle margination and its implications on intravenous anticancer drug delivery. AAPS PharmSciTech. 2014;15:762–71.
Gupta N, Kozlovskaya V, Dolmat M, Kharlampieva E. Shape recovery of spherical hydrogen-bonded multilayer capsules after osmotically induced deformation. Langmuir. 2019;35:10910–9.
Zhao YN, Sun XX, Zhang GN, Trewyn BG, Slowing II, Lin VSY. Interaction of mesoporous silica nanoparticles with human red blood cell membranes: size and surface effects. ACS Nano. 2011;5:1366–75.
Muzykantov VR, Taylor RP. Attachment of biotinylated antibody to red blood cellsantigen-binding capacity of immunoerythrocytes and their susceptibility to lysis by complement. Anal Biochem. 1994;1:142–8.
Stephan MT, Moon JJ, Um SH, Bershteyn A, Irvine DJ. Therapeutic cell engineering with surface-conjugated synthetic nanoparticles. Nat Med. 2010;16:1035–41.
Arruebo M, Valladares M, González-Fernández Á. Antibody-conjugated nanoparticles for biomedical applications. J Nanomater. 2009;2009:439389.
Zhou H, Fan Z, Li PY, Deng JJ, Arhontoulis DC, Li CY, et al. Dense and dynamic polyethylene glycol shells cloak nanoparticles from uptake by liver endothelial cells for long blood circulation. ACS Nano. 2018;12:10130–41.
Ishida T, Maeda R, Ichihara M, Irimura K, Kiwada H. Accelerated clearance of PEGylated liposomes in rats after repeated injections. J Control Release. 2003;88:35–42.
Hall SS, Mitragotri S, Daugherty PS. Identification of peptide ligands facilitating nanoparticle attachment to erythrocytes. Biotechnol Prog. 2007;23:749–54.
Lindorfe MA, Hahn CS, Foley PL, Taylor RP. Heteropolymer-mediated clearance of immune complexes via erythrocyte CR1 mechanisms and applications. Immunol Rev. 2001;183:10–24.
Zaitsev S, Spitzer D, Murciano J, Ding BS, Tliba S, Kowalska MA, et al. Sustained thromboprophylaxis mediated by an RBC-targeted pro-urokinase zymogen activated at the site of clot formation. Blood. 2010;115:5241–8.
Potempa M, Potempa J. Protease-dependent mechanisms of complement evasion by bacterial pathogens. Biol Chem. 2012;393:873–88.
Fang RH, Hu CJ, Chen KNH, Luk BT, Carpenter CW, Gao WW, et al. Lipid-insertion enables targeting functionalization of erythrocyte membrane-cloaked nanoparticles. Nanoscale. 2013;5:8884–8.
Shi G, Mukthavaram R, Kesari S, Simberg D. Distearoyl anchor-painted erythrocytes with prolonged ligand retention and circulation properties in vivo. Adv Healthc Mater. 2014;3:142–8.
Mukthavaram R, Shi G, Kesari S, Simberg D. Targeting and depletion of circulating leukocytes and cancer cells by lipophilic antibody-modified erythrocytes. J Control Release. 2014;183:146–53.
Reist CJ, Combs MJ, Croft BY, Taylor RP. Antigens pre-bound to the primate erythrocyte complement receptor via cross-linked bispecific monoclonal antibody heteropolymers are rapidly cleared from the circulation. Eur J Immunol. 1993;23:3021–7.
Zhu DM, Wu L, Suo M, Gao S, Xie W, Zan MH, et al. Engineered red blood cells for capturing circulating tumor cells with high performance. Nanoscale. 2018;10:6014–23.
Tzounakas VL, Karadimas DG, Papassideri IS, Seghatchian J, Antonelou MH. Erythrocyte-based drug delivery in Transfusion Medicine: Wandering questions seeking answers. Transfus Apher Sci. 2017;56:626–34.
Banz A, Cremel M, Mouvant A, Guerin N, Horand F, Godfrin Y. Tumor growth control using red blood cells as the antigen delivery system and poly(I:C). J Immunother. 2012;35:409–17.
Gao WW, Zhang LF. Engineering red‐blood‐cell‐membrane–coated nanoparticles for broad biomedical applications. AIChE J. 2015;61:738–46.
Murad KT, Mahany KL, Brugnara C, Kuypers FA, Eaton JW, Scott ML. Structural and functional consequences of antigenic modulation of red blood cells. Blood. 1999;93:2121–7.
Tan YX, Qiu Y, Xu H, Ji SP, Li SB, Gong F, et al. Decreased immunorejection in unmatched blood transfusions by attachment of methoxypolyethylene glycol on human red blood cells and the effect on D antigen. Transfusion. 2006;46:2122–7.
Nacharaju P, Boctor FN, Manjula BN, Acharya SA. Surface decoration of red blood cells with maleimidophenyl-polyethylene glycol facilitated by thiolation with iminothiolane: an approach to mask A, B, and D antigens to generate universal red blood cells. Transfusion. 2005;45:374–83.
Sarvi F, Najafabadi SH, Farahani EV, Shojaosadati SA. Surface coating of red blood cells with monomethoxy poly(ethylene glycol) activated with two different reagents. Iran J Chem Chem Eng-Int Engl Ed. 2008;27:1–9.
Bourgeaux V, Lanao JM, Bax BE, Godfrin YL. Drug-loaded erythrocytes: on the road toward marketing approval. Drug Des Devel Ther. 2016;10:665–76.
This work was supported by National Natural Science Foundation of China (Nos. 81872824 and 51721091). Sichuan University provided necessary services for the writing of the manuscript.
The authors declare no competing interests.
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Zhang, Sq., Fu, Q., Zhang, Yj. et al. Surface loading of nanoparticles on engineered or natural erythrocytes for prolonged circulation time: strategies and applications. Acta Pharmacol Sin 42, 1040–1054 (2021). https://doi.org/10.1038/s41401-020-00606-z
- drug delivery systems
- prolonged circulation time