表面多孔载牛血红蛋白纳米微球型血液代用品的制备及还原系统的建立  

Preparation of Bovine Hemoglobin-loaded Nanoparticles Used as Blood Substitutes and Establishment of Reduction System

作　　者：章晓兰[1] 袁媛[1] 单晓茜[1] 盛燕[1] 赵健[1] 刘昌胜[1] 

机构地区：[1]华东理工大学,教育部医用生物材料工程研究中心,上海200237

出　　处：《生物医学工程学杂志》2008年第6期1332-1337,共6页Journal of Biomedical Engineering

基　　金：十五“863”纳米专项项目资助(2004AA-302050);上海市科委纳米专项项目资助(0452nm022)

摘　　要：我们制备了表面孔径可调的载牛血红蛋白(bovine hemoglobin,BHb)纳米微球型血液代用品,并且建立一套两步还原及过程优化的非酶还原系统控制其中的高铁血红蛋白(methemoglobin,metHb)含量。用还原剂连二亚硫酸钠(sodium dithionate,SD)对氧化严重的BHb原料实行第一步还原后,进一步采用改性复乳法包埋形成纳米微球。二氯甲烷和乙腈组成的双相溶剂相对单一,二氯甲烷能有效抑制Hb的氧化,增大微球表面孔径。用不同大小分子量物质的扩散规律来估测纳米微球的表面孔径大小,由此选择血浆中的还原剂如抗坏血酸和谷胱甘肽对微球实行第二步还原。通过两步还原及过程控制,metHb含量从原料的约90%以上降为1.25%,接近天然血液中metHb含量,具备携氧/释氧性能。Bovine Hb-loaded nanoparticles with modulated size of pores, for use as blood substitutes, were prepared and a nonenzymatic reduction system including two-step-reduction and process optimization was established to control the metHb level in the present study. After the first-step-reduction procedure was performed, the raw BHb, being oxidized severely, was encapsulated to form nanoparticles by the modified double emulsion method. The binary solvent of dichloromethane （DCM） and acetonitrile （Aci） showed properties such as minimizing Hb oxidation and enlarging the pores of nanopartilces. Based on the size of pores of nanoparticles evaluated by the diffusion of various substances with different molecular weights, reducing agents such as ascorbic acid and glutathione present in the plasma were selected to perform the second-step-reduction, i.e. to further reduce the metHb in nanopartlcles. The metHb level was reduced from over 90 % in the raw materials to 1.25 % by the two-step-reduction and controlled preparation; this is near the level of native blood, possessing the ability of carrying/releasing oxygen.

关 键 词：孔径 载牛血红蛋白纳米微球 非酶还原系统 

分 类 号：R318[医药卫生—生物医学工程]