载基质细胞衍生因子-1α脂质纳米粒-声诺维复合体的制备及特性研究  

A feasibility study on preparation of SDF-1α loaded lipid nanoparticles-SonoVue compound

作　　者：曹丽娜[1] 计晓娟[1] 余更生[1] 朱旭[1] 曹阳[1] 杨海燕[1] 卢岷 何灿粲 Cao Lina;yi Xiaojuan;Yu Gengsheng;Zhu Xu;Cao Yang;Yang Haiyan;Lu Min;He Cancan.(Department of Cardiology, Children＇s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development, Chongqing Key Laboratory of Pediatrics Chongqing ＆ Institute of Ultrasound Imaging, Chongqing Medical University ＆ Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing 400014, China)

机构地区：[1]重庆医科大学附属儿童医院心脏中心儿童发育疾病研究教育部重点实验室儿科学重庆市重点实验室,400014

出　　处：《中华超声影像学杂志》2018年第5期445-451,共7页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金项目面上项目（青年基金81301300）;重庆市科委社会事业与民生保障科技创新专项（cstc2016shmszx130009）

摘　　要：目的制备一种兼具缓释作用和示踪功能的载基质细胞衍生因子-1α（SDF-1α）脂质纳米粒（SNP）-声诺维复合体（SNP—Sono Vue），并观察其致骨髓间质干细胞（BMSCs）迁移作用。方法制备SNP，检测其物理特性包括粒径大小、zeta电位、形态结构、包封率。构建SNP—SonoVue，观察SNP及低频超声（LIFU）辐照后的SNP—SonoVue缓释情况以及SNP—SonoVue中SonoVue与SNP的结合情况。分别观察以下6组的致BMSCs迁移作用以评价SNP-SonoVue的生物活性：A组，SDF-1α＋1％血清培养基；B组，SNP—SonoVue＋1％血清培养基；C组，LIFU辐照后SNP—SonoVue＋1％血清培养基；D组，空白脂质纳米粒-声诺维复合体（BNP—SonoVue）＋1％血清培养基；E组，LIFU辐照后BNP—SonoVue＋1％血清培养基；F组，PBS＋1％血清培养基（对照组）。观察SNP～SonoVue的体外显像情况。结果制得的SNP平均粒径（220．4±9．9）nm，粒径的分散指数（PDI）为0．172±0．015，平均电位（35．6±1．7）mV。透射电镜下可见SNP呈均匀分散的球形。药物包封率为96．7％，载药量为481．76ng／mg。流式细胞术分析显示SNP质量（mg）与SonoVue微泡（个）的比例为20：（2．8×10^9）～40：（2．8×10^9）是SNP—SonoVue制备的适宜条件。荧光显微镜显示复合体中的SonoVue微泡外壳有大量的DiI标记的带红色荧光SNP。药物缓释实验中可见SNP及LIFU辐照后SNP—SonoVue7d内的药物释放率分别为（68．61±3．97）0A和（63．21±5．68）％，差异无统计学意义（P〉0．05）。细胞迁移实验证实A、B、C组对BMSCs的迁移作用明显强于对照组（P〈0．（15），但A、B、C三组间的差异均无统计学意义（P〉0．05）。体外显影实验可见SNP—SonoVue复合体有明显增强显影作用。结论成功制备SNP—SonoVue，该复合体具有明显的增强显影作用和致BMSCs迁移作用。Objective To prepare SDF-1α-loaded nanoliposome （SNP）-SonoVue complex and investigate its tracing abilities, sustained-release property and effect on migration of bone marrow mesenchymal stem cells （BMSCs）. Methods The SNP was prepared to detect its physical characteristics including particle size, zeta potential, morphology, encapsulation efficiency and drug loading. SNP-SonoVue was constructed to detect the sustained release situation of SNP and SNP-SonoVue after low frequency ultrasound （LIFU） irradiation, and the connection of SNP-SonoVue was observed by fluorescence microscope. Effects of SNP-SonoVue on migration of BMSCs were detected to evaluate its bioactivity. BMSCs were divided into 6 groups , including Group A： SDF-1α ＋ 1% serum medium; Group B： SNP-SonoVue ＋ 1% serum culture medium;Group C： SNP-SonoVue ＋ 1 % serum culture medium ＋ LIFU （1 MHz, 0.5 W/cm2 , expose 30 s stop 30 s, 4 min） ; Group D： BNP-SonoVue ＋ 1 % serum medium ; Group E： BNP-SonoVue＋ 1% serum medium ＋ LIFU （1 MHz, 0.5 W/cma, expose 30 s stop 30 s, 4 min）, Group F：PBS ＋ 1% serum culture medium （control group）. Its tracing abilitie were investigated in vitro. Results The average particle size of SNP was （220.4 ± 9.9）nm, and the particle dispersion index （PDI） was （0. 172 ± 0. 015）, the average zeta potential was （35.6 ± 1.7）mv. It was showed spherical dispersion by transmission electron microscopy. The encapsulation efficiency was up to 96.7% and the drug entrapment content was 481.76 ng/mg. Flow cytometrie showed the suitable conditions for SNP-SonoVue preparation was that the ratio of SNP quality （mg） to SonoVue microbubbIes number （a） was 20：（2.8 x 10^9） to 40：（2.8 X 10^9）. Fluorescence microscopy showed that shells of SonoVue microbubbles connected with large numbers of SNP labeled with red fluorescent I）iI. Drug release experiment showed that the cumulative SDF-1α release amount of SNP and SNP-SonoVue exposed to LIFU respectively were （

关 键 词：超声疗法 脂质纳米粒 微气泡 基质细胞衍生因子1Α 骨髓间质干细胞 药物示踪 

分 类 号：R943[医药卫生—药剂学]