多柔比星磁性白蛋白纳米微粒的研制

Preparation of adriamycin magnetic albumin nanoparticles

目的:制备多柔比星磁性白蛋白纳米微粒(adriamycin magnetic albumin nanoparticles,ADM-MANs)。方法:利用人血清白蛋白、多柔比星、中位直径25 nm的Fe3O4磁粉,采用热固化交联和油剂分散法制备。光学显微镜和扫描电子显微镜观察其磁响应性和结构。应用荧光分光光度法检测磁性微粒中多柔比星含量。体外实验用MTT法检测成品微粒的细胞毒作用。结果:制备出的ADM-MANs具有磁性药物微球的形态学及物理学特征。4 000高斯磁场强度下磁响应性距离为80-100 mm。微粒直径50-400 nm,中位直径200 nm。1 mg ADM-MANs含多柔比星30．5μg,药物包封率为61．2％。其对SMMC7721肿瘤细胞的抑制率与在相应浓度下的游离多柔比星相近,半数抑制浓度分别为0．327μg·mL-1(以多柔比星含量计)和0．312μg·mL-1,统计学检验无显著性差异(P>0．05)。结论:制备出的ADM-MANs粒径小、分布均匀、磁响应性良好、药物包封率高,多柔比星细胞毒活性无明显改变。

Objective：To prepare adriamycin magnetic albumin nanoparticles（ ADM-MANs）by a modified protein denaturation technique. Methods：The ADM-MANs were prepared from human albumin, adriamycine and Fe304 powder with mean diameter of 25 nanometers using a heat denaturation and emulsion polymerization. The magnetic response and structure of ADM-MANs were analyzed by light microscope and scanning electron microscope. The loading concentration of adriamycin in the ADM-MANs was quantified by a standard spectrophotofluorometry. The cytotoxicity of the ADM-MANs was analyzed using a MTT assay in vitro. Results：The ADM-MANs featured unique microsphere shape and physical properties as of the traditional magnetic drugs. The AND-MANs had a distance of magnetic response for 80 - 100 mm under 4 000 gauss magnetic field, and a nanparticles diameter of 50 -400 nanometers（mean diameter 200 nanometer）. The loading amount of adriamycin was 30.5 microgrammes per milligram of AND-MANs, equaling to 61.2% of the inclusion rate. The inhibitory rate of ADM-MANs showed no differences from free adriamycin on SMMC7721 liver cancer cells, having the IC50 of 0. 327μg·mL^-1 versus 0. 312μg·mL^-1 （ P 〉 0.05 ）. Conclusion： This modified heat denaturation technique is feasible to prepare the AND-MANs with homogenous size, better magnetic response, higher inclusion rate and without change of cytotoxic activities.