Chen Daozhen, Tang Qiusha, Xue Wenqun, Xiang Jingying, Zhang Li, Wang Xinru
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.
J Biomed Res. 2010 Jan;24(1):26-32. doi: 10.1016/S1674-8301(10)60005-X.
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.
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.
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.
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.
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。
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。
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。
成功制备了叶酸-顺铂/HSA MNP。制备过程及相关特性数据为进一步研究奠定了基础，包括纳米粒子在体内的分布机制及其被肿瘤细胞摄取的机制。
