5-羧基荧光素标记葡聚糖Fe3 O4超顺磁性纳米颗粒的制备及性能检测  

Preparation and characteristics analysis of 5-carboxy fluorescein-dextran-coated superparamagnetic iron oxide nanoparticle

作　　者：周佳[1] 厉民[1] 夏阳 王峥[1] 郑新法[1] 黄素文[1] Zhou Jia Li Min Xia Yang Wang Zheng Zheng Xinfa Huang Suwen(Department of Gamma Knife Surgery Center, Zhejiang Provincial People＇s Hospital, Hangzhou 310014, China College of Materials Science and Engineering, Zhejiang University of Technology, Hangzhou 310014, Chin)

机构地区：[1]浙江省人民医院伽玛刀治疗中心,杭州310014 [2]浙江工业大学材料科学与工程学院,杭州310014

出　　处：《中华核医学与分子影像杂志》2017年第10期637-641,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：浙江省医药卫生科技计划（2014KYB034）

摘　　要：目的 构建5-FAM标记的葡聚糖（dextran）Fe3O4超顺磁性纳米颗粒,检测其物理和磁学性质、细胞毒性,并观察其在磁场下标记细胞的能力.方法 在超声波乳化作用下,用化学共沉淀法制备5-FAM-dextran-Fe3 O4纳米颗粒;用透射电镜和粒度分析仪观察微粒形态和大小;红外分析仪检测微粒表面结合的功能团;分光仪观察微粒的光学显像能力;振动样品磁强计检测微粒的磁响应性;MTT法检测5-FAM-dextran-Fe3 O4、dextran-Fe3 O4和Fe3 O4微粒对神经元细胞的生物毒性;共聚焦显微镜观察将荧光纳米颗粒与细胞共同孵育并给予不同外加磁场后,纳米颗粒标记神经元细胞的面积.采用单因素方差分析处理数据.结果 制得的5-FAM-dextran-Fe3 O4纳米颗粒均匀一致,直径为15~25 nm,平均粒径22 nm;其比饱和磁化强度、比剩余磁化强度、矫顽力（矫顽磁场强度）分别为86.02 A·m2·kg-1、15.05 A·m2·kg-1、5414.01 A/m,表现为超顺磁性;红外分析仪示Fe3 O4被dex-tran包裹并与5-FAM形成复合粒子;分光仪示纳米颗粒在紫外光下呈均一绿色荧光;MTT结果提示5-FAM-dextran-Fe3O4无明显细胞毒性;共聚焦显微镜下,500 mT磁场组细胞标记面积最大[（0.880±0.046）mm2;F=320.298,P〈0.05].结论 制备的5-FAM-dextran-Fe3 O4纳米颗粒可作为新型荧光-磁载体或探针应用于多种实验和临床研究.Objective To investigate the physical and magnetic properties and cytotoxity of 5-FAM-dextran-coated superparamagnetic iron oxide nanoparticles （ 5-FAM-dextran-Fe3 O4 ） , and to observe the cell-labeling character of these nanoparticles. Methods 5-FAM-dextran-Fe3 O4 were prepared by ultra-sonic and chemical coprecipitation method and the characteristics were evaluated. The size and distribution of 5-FAM-dextran-Fe3 O4 were measured by transmission electron microscope （ TEM） and Malvern Zetasizer. The organic structure of the coating was characterized by fourier translation infrared spectroscopy. The optical imaging ability was measured by ultraviolet visible spectrometer and the susceptibility was measured by vi-brating sample magnetometer. In vitro cytotoxities of 5-FAM-dextran-Fe3 O4 , dextran-Fe3 O4 and Fe3 O4 were detected by MTT assay. The area of labeled neuronal cells was observed by confocal microscopy after incuba-ted with nanoparticles under different magnetic intensities. One-way analysis of variance was used. Results The size of 5-FAM-dextran-Fe3 O4 was homogeneous under TEM, and the diameter ranged from 15 to 25 nm （ average=22 nm） by Malvern Zetasizer. The organic structure of the coating of Fe3 O4 was confirmed by fou-rier translation infrared spectroscopy. Ultraviolet visible spectrometer observation showed that the nanoparti-cles expressed unanimous green fluorescence under ultraviolet activation. The saturation magnetization, residu-al magnetization and coercivity of the nanoparticles by magnetometer detection were 86. 02 A · m2 · kg-1 , 15. 05 A·m2 ·kg-1 and 5414.01 A/m respectively, showing superparamagnetic character. MTT assay re-sults showed that 5-FAM-dextran-Fe3 O4 had no obvious cytotoxicity. The confocal microscopy observation in-dicated that the cell-labeled area reached the maximum under the magnetic field intensity of 500 mT （（0. 880±0.146） mm2, F=320.298, P〈0.05）. Conclusions 5-FAM-dextran-Fe3O4 prepared by ultra-sonic coprecipitation method have the advan

关 键 词：磁铁矿纳米颗粒 荧光素类 细胞 培养的 

分 类 号：R318.08[医药卫生—生物医学工程]