CAJ | 학술논문

目的构建肿瘤血管生成靶向的双靶点RGD10-NGR9超顺磁性氧化铁(USPIO),评价其作为活体磁共振成像(MRI)肿瘤特异性诊断制剂的能力及其成像特点.方法设计合成RGD10-NGR9双靶点杂合肽.采用共沉淀法制备网状交联葡聚糖包覆的USPIO,环氧氯丙烷活化葡聚糖,共价偶联靶向多肽,制备靶向多肽-网状交联葡聚糖-超顺磁性氧化铁复合粒子(P-CLN-Dextran-USPIO),并检测其物理性质.采用普鲁士蓝染色和邻菲罗啉比色法检测P-CLN-Dextran-USPIO体外细胞结合能力.建立裸鼠荷A549瘤模型,评价双靶点RGD10-NGR9-USPIO应用于裸鼠MRI检测的价值.结果成功制备P-CLN-Dextran-USPIO,其中Fe3 O4粒径8～ 10 nm左右,Dextran-USPIO的总粒径平均为20 nm左右,P-CLN-Dextran-USPIO的总粒径在30 nm左右,物理性质稳定.普鲁士蓝染色显示,非靶点USPIO组HUVEC细胞质内未见蓝色铁颗粒,RGD10-USPIO组和NGR9-USPIO组HUVEC细胞质内均可见蓝色铁颗粒,RGD10-NGR9-USPIO组HUVEC细胞质内蓝色铁颗粒最明显.邻菲罗啉比色显示,非靶点USPIO组、RGD10-USPIO组、NGR9-USPIO组和RGD10-NGR9-USPIO组细胞内的铁浓度分别为(1.93±0.35) μmol/L、(4.74 ±0.76) μmol/L、(5.85-0.48) μmol/L和(10.32±1.22) μmol/L,RGD10-NGR9-USPIO组细胞内的铁浓度最高,与非靶点USPIO组、RGD10-USPIO组、NGR9-USPIO组差异有统计学意义(均P＜0.05).裸鼠MRI显示,双靶点RGD10-NGR9-USPIO可以减弱肿瘤部位信号,显著提高肿瘤部位与周围组织之间的病灶对比度噪声比(CNR),较未注射时提高2.83倍(P＜0.05).结论双靶点RGD10-NGR9-USPIO作为一种阴性造影剂,具有肿瘤新生血管MRI分子成像的应用价值.
목적 구건종류혈관생성파향적쌍파점RGD10-NGR9초순자성양화철(USPIO),평개기작위활체자공진성상(MRI)종류특이성진단제제적능력급기성상특점.방법 설계합성RGD10-NGR9쌍파점잡합태.채용공침정법제비망상교련포취당포복적USPIO,배양록병완활화포취당,공개우련파향다태,제비파향다태-망상교련포취당-초순자성양화철복합입자(P-CLN-Dextran-USPIO),병검측기물이성질.채용보로사람염색화린비라람비색법검측P-CLN-Dextran-USPIO체외세포결합능력.건립라서하A549류모형,평개쌍파점RGD10-NGR9-USPIO응용우라서MRI검측적개치.결과 성공제비P-CLN-Dextran-USPIO,기중Fe3 O4립경8～ 10 nm좌우,Dextran-USPIO적총립경평균위20 nm좌우,P-CLN-Dextran-USPIO적총립경재30 nm좌우,물이성질은정.보로사람염색현시,비파점USPIO조HUVEC세포질내미견람색철과립,RGD10-USPIO조화NGR9-USPIO조HUVEC세포질내균가견람색철과립,RGD10-NGR9-USPIO조HUVEC세포질내람색철과립최명현.린비라람비색현시,비파점USPIO조、RGD10-USPIO조、NGR9-USPIO조화RGD10-NGR9-USPIO조세포내적철농도분별위(1.93±0.35) μmol/L、(4.74 ±0.76) μmol/L、(5.85-0.48) μmol/L화(10.32±1.22) μmol/L,RGD10-NGR9-USPIO조세포내적철농도최고,여비파점USPIO조、RGD10-USPIO조、NGR9-USPIO조차이유통계학의의(균P＜0.05).라서MRI현시,쌍파점RGD10-NGR9-USPIO가이감약종류부위신호,현저제고종류부위여주위조직지간적병조대비도조성비(CNR),교미주사시제고2.83배(P＜0.05).결론 쌍파점RGD10-NGR9-USPIO작위일충음성조영제,구유종류신생혈관MRI분자성상적응용개치.
Objective To construct angiogenesis-specific RGD10-NGR9 dual-targeting superparamagnetic iron oxide nanoparticles,and to evaluate its magnetic resonamce imaging (MRI) features in nude mice and potential diagnostic value in tumor MRI.Methods Dual-targeting peptides RGD10-NGR9 were designed and synthesized.Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles were synthesized by chemical co-precipitation method and the surface was modified to be hydrophilic by coating with dextran.The dual-targeting peptides RGD10-NGR9 were conjugated to USPIO.Cell binding affinity and up-taking ability of the dual-targeting USPIO nanoparticles to integrin αvβ3-APN positive cells were subsequently tested by Prussian blue staining and phenanthroline colorimetry in vitro.The RGD10-NGR9 conjugated with USPIO was injected intravenously into xenograft mice,which were scanned by MRI at predetermined time points.The MRI and contrast-to-noise ratio (CNR) values were calculated to evaluate the ability of dual-targeting USPIO as a potential contrast agent in nude mice.Results P-CLN-DextranUSPIO nanoparticles with stable physical properties were successfully constructed.The average diameter of Fe3O4 nanoparticles was 8-10 nm,that of Dextran-USPIO was about 20 nm and P-CLN-Dextran-USPIO had an average diameter about 30 nm.The in vitro studies showed a better specificity of dual-targeting USPIO nanoparticles on proliferating human umbilical vein endothelia cells (HUVEC).In vivo,RGD10-NGR9-USPIO showed a significantly reduced contrast in signal intensity and 2.83-times increased the CNR in the tumor MRIin xenograft mice.Conclusion This novel synthesized RGD10-NGR9 dual-targeting USPIO is with better specific affinity in vitro and in vivo,and might be used as a molecular contrast agent for tumor angiogenesis MRI.