Improved formulation of liposome-encapsulated hemoglobin with an anionic non-phospholipid.
Improved formulation of liposome-encapsulated hemoglobin with an anionic non-phospholipid.
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DOI：
10.1016/j.colsurfb.2009.09.038
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发表时间：
2010-02-01
影响因子：
5.8
通讯作者：
Awasthi, Vibhudutta
中科院分区：
文献类型：
--
作者：
Agashe, Hrushikesh;Lagisetty, Pallavi;Awasthi, Shanjana;Awasthi, Vibhudutta
我们正在开发脂质体包裹血红蛋白（莱赫）作为人工氧载体，用于急性失血和手术等适应症的复苏。早期尝试配制可行的莱赫遇到了规模扩大和血红蛋白含量有限的限制。在这项工作中，我们报告了一个莱赫配方含有新的阴离子非磷脂（CHHDA），提高了脂质体双层内的血红蛋白的包封效率。CHHDA是由廉价的原料以高产率合成的。通过LDH、MTT和氨基己糖苷酶测定，在体外评价该制剂，以研究对培养物中RAW 264.7巨噬细胞和HUVEC内皮细胞的细胞毒性作用。在最佳生产条件下，28摩尔%CHHDA的存在将血红蛋白含量提高到超过4 g/dL。含CHHDA的莱赫对体外培养的HUVEC和RAW细胞有一定的细胞毒作用，LDH活性最强。MTT法对两种细胞系的细胞毒性均为阴性。氨基己糖苷酶法检测，RAW细胞的增殖受到抑制，但对HUVEC细胞无明显影响。当CHHDA-LEH与分离的人血小板在体外孵育时，未观察到血小板活化。本文报道的具有新型阴离子脂质和高血红蛋白含量的莱赫制剂是对以往制剂的改进。
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 mole % 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.
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影响因子：
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