title: Improved formulation of liposome-encapsulated hemoglobin with an anionic non-phospholipid.
abstract: We are developing liposome-encapsulated hemoglobin (LEH) as an artificial oxygen carrier for resuscitation in indications, such as acute blood loss and surgery. Earlier attempts to formulate a viable LEH met with constraints of scale up and limited hemoglobin content. In this work, we report an LEH formulation containing novel anionic non-phospholipid (CHHDA) that enhances the encapsulation efficiency of hemoglobin inside the liposome bilayer. CHHDA was synthesized from inexpensive ingredients in high yields. The formulation was evaluated in vitro to investigate the cytotoxic effects on RAW 264.7 macrophages and HUVEC endothelial cells in culture by LDH, MTT and hexosaminidase assays. Under optimal conditions of manufacturing, the presence of 28 mol% of CHHDA enhanced the hemoglobin content to over 4 g/dl. The LEH containing CHHDA shows some cytotoxicity in HUVEC and RAW cells in vitro, especially by LDH assay. MTT assay was negative for cytotoxicity in both cells lines. By hexosaminidase assay, the proliferation of RAW cells, but not HUVEC cells, was inhibited. When CHHDA-LEH was incubated with isolated human platelets in vitro, no platelet activation was observed. The LEH formulation with novel anionic lipid and high hemoglobin content reported in this article is an improvement from the past preparations.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Oklahoma Health Sciences Center, 1110 North Stonewall Avenue, Oklahoma City, OK 73117, USA.
authors.forename: Hrushikesh
authors.initials: H
authors.lastname: Agashe
authors.forename: Pallavi
authors.initials: P
authors.lastname: Lagisetty
authors.forename: Shanjana
authors.initials: S
authors.lastname: Awasthi
authors.forename: Vibhudutta
authors.initials: V
authors.lastname: Awasthi
mesh_terms: D000818:Animals; D000838:Anions; D002152:Calorimetry, Differential Scanning; D016923:Cell Death / Q000187:drug effects; D004339:Drug Compounding; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D005615:Freezing; D006454:Hemoglobins / Q000737:chemistry* / Q000494:pharmacology / Q000648:ultrastructure; D006801:Humans; D008081:Liposomes / Q000737:chemistry* / Q000494:pharmacology / Q000648:ultrastructure; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D051379:Mice; D010316:Particle Size; D010743:Phospholipids / Q000737:chemistry*; D015539:Platelet Activation / Q000187:drug effects; D014470:Umbilical Cord / Q000166:cytology
chemical_list: D000838:Anions; D006454:Hemoglobins; D008081:Liposomes; D010743:Phospholipids