pH-responsive polymers for trehalose loading and desiccation protection of human red blood cells.

PP-50, a synthetic pH-responsive biopolymer, is here shown to increase the permeability of the phospholipid bilayer to trehalose, a disaccharide accumulated in desiccation tolerant organisms across all kingdoms. Uptake of 251 ± 6 mm intracellular trehalose facilitated an increase in the membrane integrity of vacuum dried cells by a factor of 9 ± 1 and reduced extent of hemoglobin oxidation in dried cells from 66 ± 1% to 23 ± 3%. To elucidate the mechanism of PP-50 mediated trehalose delivery, permeability studies were conducted using molecules ranging in size from sucrose to 10 kDa poly(ethylene glycol). It was shown that the logarithm of relative diffusant membrane permeability decreased linearly with diffusant molecular volume, suggesting transport via non-Stokesian diffusion. Consistent with this conclusion, topographic atomic force micrographs reported membrane thinning proximate to PP-50 adsorption on the erythrocyte membrane, a phenomenon associated with increased incidence of phospholipid hydrocarbon chain bending.

[1]  S. Ludtke,et al.  Membrane thinning caused by magainin 2. , 1995, Biochemistry.

[2]  F. Devínsky,et al.  Interaction of surfactants with model and biological membranes. II. Effect of N-alkyl-N,N,N-trimethylammonium ions on phosphatidylcholine bilayers as studied by spin probe ESR. , 1990, Chemistry and physics of lipids.

[3]  B. Bechinger,et al.  Detergent-like actions of linear amphipathic cationic antimicrobial peptides. , 2006, Biochimica et biophysica acta.

[4]  H. Träuble,et al.  The movement of molecules across lipid membranes: A molecular theory , 1971, The Journal of Membrane Biology.

[5]  B. Tighe,et al.  Responsive hydrophobically associating polymers: a review of structure and properties. , 2001, Advanced drug delivery reviews.

[6]  H Denny Kamaruddin,et al.  Some observations about the application of Fick's first law for membrane separation of multicomponent mixtures , 1997 .

[7]  J. Crowe Trehalose as a "chemical chaperone": fact and fantasy. , 2007, Advances in experimental medicine and biology.

[8]  Z. Yue,et al.  Synthesis and pH-responsive properties of pseudo-peptides containing hydrophobic amino acid grafts , 2009 .

[9]  A. Bondi van der Waals Volumes and Radii , 1964 .

[10]  Z. Török,et al.  Loading red blood cells with trehalose: a step towards biostabilization. , 2004, Cryobiology.

[11]  R. Benesch,et al.  Equations for the spectrophotometric analysis of hemoglobin mixtures. , 1973, Analytical biochemistry.

[12]  S. Marčelja,et al.  Physical principles of membrane organization , 1980, Quarterly Reviews of Biophysics.

[13]  Rongjun Chen,et al.  Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival. , 2010, Biomaterials.

[14]  Z. Török,et al.  Preservation of Trehalose-Loaded Red Blood Cells by Lyophilization , 2005 .

[15]  J. Acker,et al.  Biopreservation of red blood cells – the struggle with hemoglobin oxidation , 2010, The FEBS journal.

[16]  W. R. Lieb,et al.  Non-stokesian nature of transverse diffusion within human red cell membranes , 2005, The Journal of Membrane Biology.

[17]  A. Hoffman,et al.  Design of "smart" nano-scale delivery systems for biomolecular therapeutics , 2007 .

[18]  Z. Yue,et al.  Modulation of the pH-responsive properties of poly(L-lysine iso-phthalamide) grafted with a poly(ethylene glycol) analogue. , 2005, Biomaterials.

[19]  M. B. Banaszak Holl,et al.  Membrane thinning due to antimicrobial peptide binding: an atomic force microscopy study of MSI-78 in lipid bilayers. , 2005, Biophysical journal.

[20]  R M Hochmuth,et al.  Mechanical measurement of red cell membrane thickness. , 1983, Science.

[21]  C. M. Peterson,et al.  Imaging cells with the atomic force microscope. , 1990, Journal of structural biology.

[22]  M. Lafleur,et al.  Characterization of the membrane-destabilizing properties of different pH-sensitive methacrylic acid copolymers. , 2003, Biochimica et biophysica acta.

[23]  Huey W. Huang,et al.  Action of antimicrobial peptides: two-state model. , 2000, Biochemistry.

[24]  E. M. Renkin,et al.  FILTRATION, DIFFUSION, AND MOLECULAR SIEVING THROUGH POROUS CELLULOSE MEMBRANES , 1954, The Journal of general physiology.

[25]  Baolin Liu,et al.  Effects of glycerol pretreatment on recovery and antioxidant enzyme activities of lyophilized red blood cells. , 2008, Cryo letters.

[26]  J. Acker,et al.  Biopreservation of red blood cells: past, present, and future. , 2005, Transfusion medicine reviews.

[27]  Baolin Liu,et al.  Loading trehalose into red blood cells by electroporation and its application in freeze-drying. , 2010, Cryo letters.

[28]  C. Foged,et al.  Cell-penetrating peptides for drug delivery across membrane barriers , 2008, Expert opinion on drug delivery.

[29]  J. Acker,et al.  Effect of Liposome Charge and Composition on the Delivery of Trehalose into Red Blood Cells , 2008 .

[30]  A. Engel,et al.  Atomic force microscopy of biological membranes. , 2009, Biophysical journal.

[31]  R. Benz,et al.  Properties of the leak permeability induced by a cytotoxic protein from Pseudomonas aeruginosa (PACT) in rat erythrocytes and black lipid membranes. , 1985, Biochimica et biophysica acta.

[32]  H. Ginsburg,et al.  Biophysical analysis of novel transport pathways induced in red blood cell membranes , 2005, The Journal of Membrane Biology.

[33]  Comparison of frozen versus desiccated reference human red blood cells for hemagglutination assays , 2009, Transfusion.

[34]  C. Haest,et al.  Formation of aqueous pores in the human erythrocyte membrane after oxidative cross-linking of spectrin by diamide. , 1983, Biochimica et biophysica acta.

[35]  Rongjun Chen,et al.  Aqueous solution behaviour and membrane disruptive activity of pH-responsive PEGylated pseudo-peptides and their intracellular distribution. , 2008, Biomaterials.

[36]  Y. Ikarashi,et al.  Haemolysis of various mammalian erythrocytes in sodium chloride, glucose and phosphate-buffer solutions , 1979, Laboratory animals.

[37]  M. Toner,et al.  Long-term storage of tissues by cryopreservation: critical issues. , 1996, Biomaterials.

[38]  J. Acker,et al.  Mechanism of hemoglobin-induced cellular injury in desiccated red blood cells. , 2010, Free radical biology & medicine.

[39]  Rongjun Chen,et al.  The role of hydrophobic amino acid grafts in the enhancement of membrane-disruptive activity of pH-responsive pseudo-peptides. , 2009, Biomaterials.

[40]  M. Al‐Rubeai,et al.  Retroviral vectors for human gene delivery. , 2002, Biotechnology advances.

[41]  J. Crowe Trehalose As a “Chemical Chaperone” , 2007 .

[42]  H. Bayley,et al.  Beneficial effect of intracellular trehalose on the membrane integrity of dried mammalian cells. , 2001, Cryobiology.

[43]  Roberto D Lins,et al.  Interaction of the disaccharide trehalose with a phospholipid bilayer: a molecular dynamics study. , 2004, Biophysical journal.

[44]  Songmiao Liang,et al.  Transport of Glucose and Poly(ethylene glycol)s in Agarose Gels Studied by the Refractive Index Method , 2005 .

[45]  M. Clemens,et al.  Lipid peroxidation in erythrocytes. , 1987, Chemistry and physics of lipids.

[46]  N. Fullwood,et al.  The use of trehalose-treated freeze-dried amniotic membrane for ocular surface reconstruction. , 2008, Biomaterials.

[47]  J. Crowe,et al.  Preservation of freeze-dried liposomes by trehalose. , 1985, Archives of biochemistry and biophysics.

[48]  M. Eccleston,et al.  pH-responsive pseudo-peptides for cell membrane disruption. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[49]  S. White,et al.  'Detergent-like' permeabilization of anionic lipid vesicles by melittin. , 2001, Biochimica et biophysica acta.