Centis Valérie, Vermette Patrick
Laboratoire de Bioingénierie et de Biophysique de l'Université de Sherbrooke, Department of Chemical Engineering, Université de Sherbrooke, 2500, boul. de l'Université, Sherbrooke (QC), Canada J1K 2R1.
Colloids Surf B Biointerfaces. 2008 Sep 1;65(2):239-46. doi: 10.1016/j.colsurfb.2008.04.009. Epub 2008 Apr 24.
PEGylated liposomes encapsulating human hemoglobin as oxygen carriers were prepared from purified carbonylhemoglobin (HbCO) solution and a lipid mixture composed of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), cholesterol, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[poly(ethylene glycol) 2000] (DMPE-PEG(2000)) and palmitic acid. Hemoglobin was extracted and purified from human blood samples. SDS-PAGE was used to assess its purity. Diameter of liposomes containing hemoglobin was controlled to approximately 200 nm using extrusion as measured by dynamic light scattering and transmission electron microscopy. Liposome size distributions were shown to remain unimodal over 14 days, even at different storage temperatures. Zeta potential measurements revealed that liposome containing hemoglobin have a net surface charge of -7.16+/-0.33 mV. Also, hemoglobin encapsulated in liposomes was able to perform several cycles of oxygen loading and unloading using oxygen (O(2)) and carbon monoxide (CO). The hemoglobin vesicle dispersion showed some toxicity as revealed by three in vitro assays in which endothelial cell (HUVECs) monolayers were exposed to these dispersions. Cytotoxicity was function of the liposome concentration in the culture medium.
以纯化的羰基血红蛋白（HbCO）溶液和由1,2-二棕榈酰-sn-甘油-3-磷脂酰胆碱（DPPC）、胆固醇、1,2-二肉豆蔻酰-sn-甘油-3-磷酸乙醇胺-N-[聚（乙二醇）2000]（DMPE-PEG(2000)）和棕榈酸组成的脂质混合物制备了包裹人血红蛋白作为氧载体的聚乙二醇化脂质体。从人血样本中提取并纯化血红蛋白。使用SDS-PAGE评估其纯度。通过动态光散射和透射电子显微镜测量，采用挤压法将含血红蛋白的脂质体直径控制在约200nm。结果表明，即使在不同的储存温度下，脂质体大小分布在14天内仍保持单峰。ζ电位测量显示，含血红蛋白的脂质体净表面电荷为-7.16±0.33mV。此外，包裹在脂质体中的血红蛋白能够利用氧气（O(2)）和一氧化碳（CO）进行多个循环的氧加载和卸载。三种体外试验表明，内皮细胞（HUVECs）单层暴露于这些分散体时，血红蛋白囊泡分散体具有一定毒性。细胞毒性是培养基中脂质体浓度的函数。
