Surface-modified loaded human red blood cells for targeting and delivery of drugs

Red blood cells (RBCs) are natural carriers which can be used for targeted drug delivery. Conditions during loading and surface modification are essential for carrier-RBC preparation for specifically targeted drug delivery. Therefore, human RBCs were loaded with albumin and magnetic nanoparticles (NPs) by different hypotonic haemolysis procedures and compared based on loading efficiency and membrane damage. Samples were analysed by flow cytometry and confocal microscopy. The optimized loading procedure resulted in 90% albumin-loaded carrier-RBCs with <4% Annexin V binding and 263 pg iron per RBC after loading with iron oxide NPs. Albumin-loaded RBCs were subsequently surface conjugated with insulin and IgG via biotin–streptavidin. Insulin-conjugated carrier-RBCs were observed to attach and to be internalized by cultured endothelial cells. Uptake was not observed for carrier-RBCs non-specifically modified with IgG. Attachment of other peptides with high specificity will open novel opportunities for targeting various cells, tissues and for crossing biological barriers.

[1]  J. Estaquier,et al.  Cellular and molecular mechanisms of senescent erythrocyte phagocytosis by macrophages. A review. , 1998, Biochimie.

[2]  Yu-Hsin Lin,et al.  Preparation and characterization of nanoparticles shelled with chitosan for oral insulin delivery. , 2007, Biomacromolecules.

[3]  V. Muzykantov,et al.  Avidin-induced lysis of biotinylated erythrocytes by homologous complement via the alternative pathway depends on avidin's ability of multipoint binding with biotinylated membrane. , 1992, Biochimica et biophysica acta.

[4]  M. Dathe,et al.  Dipalmitoylation of a cellular uptake-mediating apolipoprotein E-derived peptide as a promising modification for stable anchorage in liposomal drug carriers. , 2006, Biochimica et biophysica acta.

[5]  M. Magnani,et al.  Cell-based drug delivery. , 2008, Advanced drug delivery reviews.

[6]  D. Daleke Regulation of transbilayer plasma membrane phospholipid asymmetry Published, JLR Papers in Press, December 16, 2002. DOI 10.1194/jlr.R200019-JLR200 , 2003, Journal of Lipid Research.

[7]  W. Duckworth,et al.  Human red blood cell insulin-degrading enzyme and rat skeletal muscle insulin protease share antigenic sites and generate identical products from insulin. , 1990, The Journal of biological chemistry.

[8]  M. Magnani,et al.  Targeting antiretroviral nucleoside analogues in phosphorylated form to macrophages: in vitro and in vivo studies. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[9]  G P Samokhin,et al.  Targeting of enzyme immobilized on erythrocyte membrane to collagen‐coated surface , 1985, FEBS letters.

[10]  R. Georgieva,et al.  Fabrication of Colloidal Stable, Thermosensitive, and Biocompatible Magnetite Nanoparticles and Study of Their Reversible Agglomeration in Aqueous Milieu , 2009 .

[11]  Vladimir R Muzykantov,et al.  Drug delivery by red blood cells: vascular carriers designed by mother nature , 2010, Expert opinion on drug delivery.

[12]  Mehrdad Hamidi,et al.  Applications of carrier erythrocytes in delivery of biopharmaceuticals. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[13]  V R Muzykantov,et al.  Enhanced complement susceptibility of avidin-biotin-treated human erythrocytes is a consequence of neutralization of the complement regulators CD59 and decay accelerating factor. , 1995, The Biochemical journal.

[14]  V. Muzykantov,et al.  Avidin attachment to biotinylated erythrocytes induces homologous lysis via the alternative pathway of complement. , 1991, Blood.

[15]  Silvia Muro,et al.  Advanced drug delivery systems that target the vascular endothelium. , 2006, Molecular interventions.

[16]  P. Hinderling,et al.  Red blood cells: a neglected compartment in pharmacokinetics and pharmacodynamics. , 1997, Pharmacological reviews.

[17]  P. Hochstein,et al.  Aging human erythrocytes. Differential sensitivity of young and old erythrocytes to hemolysis induced by peroxide in the presence of thyroxine. , 1976, Archives of biochemistry and biophysics.

[18]  J. Carpentier,et al.  Biosynthesis and regulation of the insulin receptor. , 1989, The Yale journal of biology and medicine.

[19]  L. H. Reddy Drug delivery to tumours: recent strategies , 2005, The Journal of pharmacy and pharmacology.

[20]  D. Stasiw,et al.  Erythrocyte fragility in aging. , 1974, Biochimica et biophysica acta.

[21]  G. Ihler,et al.  An erythrocyte encapsulator dialyzer used in preparing large quantities of erythrocyte ghosts and encapsulation of a pesticide in erythrocyte ghosts. , 1980, Analytical biochemistry.

[22]  J. Kreuter,et al.  Covalent attachment of apolipoprotein A-I and apolipoprotein B-100 to albumin nanoparticles enables drug transport into the brain. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[23]  V. Muzykantov,et al.  Red blood cell targeting to smooth muscle cells , 1986, FEBS letters.

[24]  G. King,et al.  Receptor-mediated transport of insulin across endothelial cells. , 1985, Science.

[25]  F. Lang,et al.  Suicidal Erythrocyte Death Following Cellular K+ Loss , 2006, Cellular Physiology and Biochemistry.

[26]  M. Magnani,et al.  Erythrocyte-based drug delivery , 2005, Expert opinion on drug delivery.

[27]  M. Magnani,et al.  Erythrocyte-mediated delivery of drugs, peptides and modified oligonucleotides , 2002, Gene Therapy.

[28]  M. Tonetti,et al.  Carrier Erythrocytes , 1993, Clinical Pharmacokinetics.

[29]  Samir Mitragotri,et al.  Prolonged circulation of large polymeric nanoparticles by non-covalent adsorption on erythrocytes. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[30]  B. Trump,et al.  Membrane Alterations in Hemolysis: Internalization of Plasmalemma Induced by Primaquine , 1969, Science.

[31]  V. Muzykantov,et al.  Avidin attachment to red blood cells via a phospholipid derivative of biotin provides complement-resistant immunoerythrocytes. , 1993, Journal of immunological methods.

[32]  R. Mylvaganam,et al.  Use of platelets as drug carriers for the treatment of hematologic diseases. , 1987, Methods in enzymology.

[33]  A. Zanella,et al.  Erythrocyte engineering for drug delivery and targeting , 1998, Biotechnology and applied biochemistry.

[34]  P. Hochstein,et al.  Aging of human erythrocytes , 1976 .

[35]  J. Weisel,et al.  The spatial dynamics of fibrin clot dissolution catalyzed by erythrocyte‐bound vs. free fibrinolytics , 2010, Journal of thrombosis and haemostasis : JTH.

[36]  D. Hirst,et al.  Mesenchymal stem cells as a gene therapy carrier for treatment of fibrosarcoma. , 2009, Cytotherapy.

[37]  T. Allen Ligand-targeted therapeutics in anticancer therapy , 2002, Nature Reviews Cancer.

[38]  U Teichgräber,et al.  Magnetite-loaded carrier erythrocytes as contrast agents for magnetic resonance imaging. , 2006, Nano letters.

[39]  V. Muzykantov,et al.  Regulation of the complement-mediated elimination of red blood cells modified with biotin and streptavidin. , 1996, Analytical biochemistry.

[40]  M. Weiner,et al.  Incorporation of inositol hexaphosphate into red blood cells mediated by dimethyl sulfoxide. , 1983, Life sciences.

[41]  Mehrdad Hamidi,et al.  Carrier Erythrocytes: An Overview , 2003, Drug delivery.

[42]  A. Voigt,et al.  In vitro inhibition of fungal activity by macrophage-mediated sequestration and release of encapsulated amphotericin B nanosupension in red blood cells. , 2010, Small.

[43]  V. Muzykantov,et al.  Directed targeting of immunoerythrocytes provides local protection of endothelial cells from damage by hydrogen peroxide. , 1987, The American journal of pathology.

[44]  M. Magnani,et al.  Red blood cell-mediated delivery of recombinant HIV-1 Tat protein in mice induces anti-Tat neutralizing antibodies and CTL. , 2003, Vaccine.

[45]  M. Magnani,et al.  Comparison of uricase-bound and uricase-loaded erythrocytes as bioreactors for uric acid degradation. , 1992, Advances in experimental medicine and biology.

[46]  H. Bäumler,et al.  Influence of 30 Gy Gamma Irradiation on the Quality of Red Blood Cell Concentrates in Several Storage Media , 1999, Transfusion Medicine and Hemotherapy.

[47]  V R Muzykantov,et al.  Attachment of biotinylated antibody to red blood cells: antigen-binding capacity of immunoerythrocytes and their susceptibility to lysis by complement. , 1994, Analytical biochemistry.

[48]  T. Tsong,et al.  Formation and resealing of pores of controlled sizes in human erythrocyte membrane , 1977, Nature.

[49]  Han‐Chung Wu,et al.  Peptide-Mediated Liposomal Drug Delivery System Targeting Tumor Blood Vessels in Anticancer Therapy , 2010, Journal of oncology.

[50]  M. Magnani,et al.  Red blood cells as an antigen‐delivery system , 1992, Biotechnology and applied biochemistry.