叶酸偶联人血清白蛋白磁性顺铂纳米粒的制备与表征

The preparation and characterization of folate-conjugated human serum albumin magnetic cisplatin nanoparticles.

作者信息

Chen Daozhen, Tang Qiusha, Xue Wenqun, Xiang Jingying, Zhang Li, Wang Xinru

机构信息

Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing, 210042, China ; Clinical Laboratory ,Wuxi Hospital for Maternal and Child Health Care Affiliated Nanjing Medical University, Wuxi, Jiangsu,214002, China.

出版信息

J Biomed Res. 2010 Jan;24(1):26-32. doi: 10.1016/S1674-8301(10)60005-X.

DOI:10.1016/S1674-8301(10)60005-X

PMID:23554608

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3596532/

Abstract

OBJECTIVE

Nanoparticles are becoming an important method of targeted drug delivery. To evaluate the importance of folate-conjugated human serum albumin (HSA) magnetic nanoparticles (Folate-CDDP/HSA MNP), we prepared drug-loaded Folate-CDDP/HSA MNPs and characterized their features.

METHODS

First, folate was conjugated with HSA under the effect of a condensing agent, and the conjugating rate was evaluated by a colorimetric method using 2, 4, 6 - trinitrobenzene sulfonic acid. Second, under N2 gas, Fe3O4 magnetic nanomaterials were prepared and characterized by using transmission electron microscopy (TEM), SEM-EDS and X-ray diffraction (XRD). Finally, Folate-CDDP/HSA MNP was prepared by using a solvent evaporation technique. TEM was used to observe particle morphology. The particle size and distribution of the prepared complexes were determined by a Laser particle size analyzer. Drug loading volume and drug release were investigated by a high performance liquid chromatography method (HPLC) in vitro.

RESULTS

We successfully prepared folate-conjugated HSA and its conjugating rate was 27.26 µg/mg. Under TEM, Fe3O4 magnetic nanoparticles were highly electron density and had an even size distribution in the range of 10-20 nm. It was confirmed by SEM-EDS and XRD that Fe3O4 magnetic nanoparticles had been successfully prepared. Under TEM, drug-loaded magnetic nanoparticles were observed, which had a round shape, similar uniform size and smooth surface. Their average size was 79 nm which was determined by laser scattering, and they exhibited magnetic responsiveness. Encapsulation efficiency was 89.75% and effective drug loading was calculated to be 15.25%. The release results in vitro showed that the half release time (t½) of cisplatin in cisplatin Solution and Folate-CDDP/HSA MNP was 65 min and 24 h respectively, which indicated that microspheres had an obvious effect of sustained-release.

CONCLUSION

Folate-CDDP/HSA MNPs were prepared successfully. The preparation process and related characteristics data provided a foundation for further study, including the mechanism of the nanoparticles distribution in vivo and their intake by tumor cells.

摘要

目的

纳米粒子正成为靶向给药的一种重要方法。为评估叶酸偶联人血清白蛋白（HSA）磁性纳米粒子（叶酸-顺铂/HSA MNP）的重要性，我们制备了载药的叶酸-顺铂/HSA MNP并对其特性进行了表征。

方法

首先，在缩合剂作用下将叶酸与HSA偶联，采用2,4,6-三硝基苯磺酸比色法评估偶联率。其次，在氮气氛围下制备Fe3O4磁性纳米材料，并通过透射电子显微镜（TEM）、扫描电子显微镜-能谱仪（SEM-EDS）和X射线衍射（XRD）对其进行表征。最后，采用溶剂蒸发技术制备叶酸-顺铂/HSA MNP。用TEM观察颗粒形态。用激光粒度分析仪测定所制备复合物的粒径和分布。采用高效液相色谱法（HPLC）体外研究载药量和药物释放情况。

结果

我们成功制备了叶酸偶联的HSA，其偶联率为27.26 μg/mg。在TEM下，Fe3O4磁性纳米粒子具有高电子密度，粒径在10 - 20 nm范围内分布均匀。通过SEM-EDS和XRD证实成功制备了Fe3O4磁性纳米粒子。在TEM下观察到载药磁性纳米粒子，呈圆形，粒径相似且均匀，表面光滑。通过激光散射测定其平均粒径为79 nm，且表现出磁响应性。包封率为89.75%，有效载药量经计算为15.25%。体外释放结果表明，顺铂溶液和顺铂-Folate-CDDP/HSA MNP中顺铂的半衰期（t½）分别为65分钟和24小时，这表明微球具有明显的缓释作用。