title: Biological evaluation of liposome-encapsulated hemoglobin surface-modified with a novel PEGylated nonphospholipid amphiphile.
abstract: Traumatic injury is often associated with hemorrhagic shock. Liposome-encapsulated hemoglobin (LEH) is being developed as an artificial oxygen carrier to address post-hemorrhage oxygen and volume deficit. Here, we report a new composition of LEH based on the use of polyethylene glycol (PEG2K ) conjugated with nonphospholipid hexadecylcarbamoylmethylhexadecanoate (HDAS) to modify the surface of LEH particles. LEH was manufactured by the high-pressure homogenization method using dipalmitoylphosphatidylcholine (∼38 mol%), cholesterol (∼38 mol%), HDAS (∼20 mol%), and highly purified stroma-free human hemoglobin. HDAS-PEG2K was postinserted into the resultant LEH to generate HDAS-PEG2K -LEH. We investigated the potential immune response to HDAS-PEG2K -LEH in a mice model. At the same time, the preparation was tested in a rat model to study the effect of repeated HDAS-PEG2K -LEH injection over 4 weeks. We found that HDAS-PEG2K modification substantially reduced the circulating levels of anaphylatoxins C3a and C5a, as well as plasma levels of thromboxane B2, in mice. Repeated injections of HDAS-PEG2K -LEH in rats did not appear to alter its clearance profile after 4 weeks of treatment. No antibody response against human hemoglobin or PEG was detected in rat plasma. Histological observations of lung, liver, spleen, and kidney were not significantly different between saline-treated rats and HDAS-PEG2K -LEH-treated rats. Immunohistochemical staining for rat heme oxygenase-1 (HO-1) did not show induced expression of HO-1 in these organs. These results suggest that the new surface modification of LEH is immune-neutral and does not adversely affect histology even after repeated administration.
authors.affiliation: Department of Pharmaceutical Sciences, Research Imaging Facility, University of Oklahoma Health Science Center, Oklahoma City, OK, USA.
authors.forename: Vivek R
authors.initials: VR
authors.lastname: Yadav
authors.forename: Okhil
authors.initials: O
authors.lastname: Nag
authors.forename: Vibhudutta
authors.initials: V
authors.lastname: Awasthi
mesh_terms: D000818:Animals; D001802:Blood Substitutes; D016503:Drug Delivery Systems; D006454:Hemoglobins; D008081:Liposomes; D008297:Male; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D012771:Shock, Hemorrhagic / Q000628:therapy*
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D008081:Liposomes; C016484:hemoglobin, stroma free
keywords: Complement; Liposome-encapsulated hemoglobin; Oxygen delivery
references.citation: J Pharmacol Exp Ther. 2002 Mar;300(3):976-83
references.pmid: 11861806
references.citation: Artif Cells Blood Substit Immobil Biotechnol. 1994;22(3):503-16
references.pmid: 7994372
references.citation: Transfusion. 1997 Feb;37(2):150-9
references.pmid: 9051089
references.citation: Colloids Surf B Biointerfaces. 2010 Feb 1;75(2):573-83
references.pmid: 19896346
references.citation: Mol Immunol. 1999 Sep-Oct;36(13-14):843-52
references.pmid: 10698338
references.citation: J Pharmacol Exp Ther. 2004 Apr;309(1):241-8
references.pmid: 14718581
references.citation: Eur J Pharm Sci. 2013 May 13;49(2):265-71
references.pmid: 23528740
references.citation: J Am Coll Surg. 2009 Apr;208(4):592-8
references.pmid: 19476795
references.citation: Biotechnol Prog. 1999 Jul-Aug;15(4):655-66
references.pmid: 10441357
references.citation: Adv Exp Med Biol. 1988;238:161-70
references.pmid: 3250239
references.citation: Artif Cells Blood Substit Immobil Biotechnol. 2008;36(1):34-44
references.pmid: 18293159
references.citation: Thromb Haemost. 1990 Aug 13;64(1):172-6
references.pmid: 2274922
references.citation: J Trauma. 2006 Jun;60(6 Suppl):S3-11
references.pmid: 16763478
references.citation: Biochem Biophys Res Commun. 1994 Nov 30;205(1):255-63
references.pmid: 7999033
references.citation: J Nutr. 1995 Jun;125(6 Suppl):1661S-1665S
references.pmid: 7782922
references.citation: J Pharmacol Exp Ther. 2004 Dec;311(3):874-84
references.pmid: 15297471
references.citation: J Liposome Res. 2002 Feb-May;12(1-2):165-72
references.pmid: 12604051
references.citation: J Biomed Mater Res. 1995 Feb;29(2):189-96
references.pmid: 7738065
references.citation: Methods Enzymol. 2003;373:136-54
references.pmid: 14714402
references.citation: Anal Biochem. 1980 May 1;104(1):10-4
references.pmid: 6892980
references.citation: Int J Pharm. 2013 Mar 25;446(1-2):119-29
references.pmid: 23419666
references.citation: FASEB J. 2006 Dec;20(14):2591-3
references.pmid: 17065229
references.citation: Exp Hematol. 1997 Dec;25(13):1347-56
references.pmid: 9406994
references.citation: Biochem Biophys Res Commun. 1986 Dec 15;141(2):528-33
references.pmid: 3099789
references.citation: Adv Drug Deliv Rev. 1998 Jun 8;32(1-2):19-29
references.pmid: 10837633
references.citation: Int J Pharm. 2004 Sep 28;283(1-2):53-62
references.pmid: 15363501
references.citation: Chem Phys Lipids. 1986 Jun-Jul;40(2-4):359-72
references.pmid: 3742678
references.citation: J Immunol. 1991 Jun 15;146(12):4234-41
references.pmid: 2040798
references.citation: Curr Drug Deliv. 2005 Apr;2(2):133-42
references.pmid: 16305414
references.citation: Ann Surg. 2007 Jun;245(6):931-42
references.pmid: 17522519
references.citation: Mol Immunol. 2008 Oct;45(16):4073-9
references.pmid: 18706699
references.citation: Artif Organs. 2012 Feb;36(2):194-201
references.pmid: 21790678
references.citation: Artif Cells Blood Substit Immobil Biotechnol. 1996 Nov;24(6):643-53
references.pmid: 8922233
references.citation: Artif Cells Blood Substit Immobil Biotechnol. 1997 Jul;25(4):347-55
references.pmid: 9242930
references.citation: J Control Release. 2007 Oct 8;122(3):349-55
references.pmid: 17610982
references.citation: Surgery. 2007 Aug;142(2):289-94
references.pmid: 17689698
references.citation: Artif Organs. 2009 Feb;33(2):169-76
references.pmid: 19178463
references.citation: J Clin Invest. 1998 Feb 1;101(3):604-12
references.pmid: 9449694
references.citation: Vaccine. 1992;10(3):151-8
references.pmid: 1557930
references.citation: Langmuir. 2007 Jul 3;23(14):7695-9
references.pmid: 17564471
references.citation: J Pharmacol Exp Ther. 1999 Feb;288(2):665-70
references.pmid: 9918573
references.citation: Vaccine. 1996 Feb;14(3):219-25
references.pmid: 8920703
references.citation: Biotechnol Prog. 1996 Jan-Feb;12(1):119-25
references.pmid: 8845102
references.citation: Biol Pharm Bull. 1997 Nov;20(11):1208-11
references.pmid: 9401734