摘要
目的制备一种搭载酞菁铁(Iron(Ⅱ)phthalocyanine,FePc)并利用适配体AS1411引导靶向人乳腺癌MCF-7细胞的新型纳米粒(AS1411-PLGA@FePc@PFP)。体外评价其理化性质、对MCF-7细胞的靶向能力、光声/超声的多模态成像能力、光热性能和光致相变及体内多模态成像。方法基于双步超声乳化法制备搭载FePc的非靶向纳米粒(PLGA@FePc@PFP),运用碳二亚胺法将尾部润饰有NH2的适配体AS1411与PLGA@FePc@PFP连接,制备出靶向纳米粒(AS1411-PLGA@FePc@PFP);检测其基本理化性质;激光共聚焦与流式细胞仪用于评估AS1411与PLGA@FePc@PFP连接情况;运用CCK-8检测AS1411-PLGA@FePc@PFP的细胞毒性;用激光共聚焦观察AS1411-PLGA@FePc@PFP体外靶向能力;分别用光声和超声成像检测AS1411-PLGA@FePc@PFP在体内外的成像;用近红外激光(660 nm)、热成像仪检测其体内外光热转化能力,用光镜记录激光辐照前后纳米粒的变化。结果成功制备出AS1411-PLGA@FePc@PFP,透射电镜下观察呈圆球形,大小均匀,粒径(208.03±8.96)nm;电位(-10.8±1.14)mV;FePc包封率为(83.96±1.61)%。CCK-8检测结果显示不同浓度的AS1411-PLGA@FePc@PFP未见明显的细胞毒性;流式细胞仪检测AS1411已成功连接于纳米粒,其连接率为(93.31±2.58)%;激光共聚焦显示在靶向组中MCF-7细胞周围的纳米粒明显多于非靶向组。经过激光辐照后,带有FePc的纳米粒温度迅速升高,光镜下观察纳米粒发生相变,因此其具备良好的光热转化能力。光声成像结果显示:与不具备FePc的纳米粒相比,AS1411-PLGA@FePc@PFP具有良好的光声信号,并且纳米粒浓度越高,其信号强度越强,呈线性相关。超声和造影结果显示与不具备FePc的纳米粒相比,AS1411-PLGA@FePc@PFP的FePc可以实现良好光热转化,用于增强超声造影效果,且纳米粒浓度越高,成像效果越明显,呈线性相关。体内光声成像显示,尾静脉注射AS1411-PLGA@FePc@PFP和PLGA@FePc@PFP 6h后移植瘤内的光声信号�
Objective To prepare a novel nanoparticle carrying iron(Ⅱ)phthalocyanine(FePc)coupled with aptamer AS1411 for targeting human breast cancer MCF-7 cells and evaluate its physicochemical properties,targeting ability to MCF-7 cells,photoacoustic/ultrasonic multimodal imaging capabilities,photothermal performance,photoinduced phase transitions in vitro and multimodal imaging performance in vivo.Methods FePc-encapsulated non-targeted nanoparticles(PLGA@FePc@PFP)were prepared by a two-step phacoemulsification method,and the targeted nanoparticles(AS1411-PLGA@FePc@PFP NPs)were prepared by combining the aptamer AS1411 modified with NH2 and the PLGA@FePc@PFP nanoparticles using the carbodiimide method.The physicochemical properties of AS1411-PLGA@FePc@PFP NPs were detected,and laser confocal microscopy and flow cytometry were used to evaluate the binding between AS1411 and the nanoparticles.CCK-8 assay was used to detect the cytotoxicity of AS1411-PLGA@FePc@PFP;laser confocal microscopy was used to observe the in vitro targeting ability of the nanoparticles;photoacoustic and ultrasonic imagers were used to test their imaging performance and targeting ability both in vitro and in vivo.Near-infrared laser(660 nm)and thermal imagers were used to test the in vitro photothermal conversion abilities of the nanoparticles,and the changes of the nanoparticles after laser irradiation were observed under an optical microscope.The temperature change of AS1411-PLGA@FePc@PFP in tumor-bearing nude mice was detected in response to 660 nm laser irradiation.Results AS1411-PLGA@FePc@PFP was successfully prepared,which had an average diameter of 208.03±8.96 nm with a narrow particle size distribution,a mean zeta potential of-10.8±1.14 mV,and a FePc encapsulation rate of(83.96±1.61)%.The results of CCK-8 assay showed that the nanoparticles at varying concentrations had no obvious cytotoxicity.Flow cytometry showed an efficient binding of AS1411 with the nanoparticles had a binding rate of(93.31±2.58)%.Laser confocal analysis showed that
作者
何雨蓓
郝兰
李倩茹
黄美盈
熊亚岚
郑晨曦
王志刚
袁耿彪
HE Yubei;HAO Lan;LI Qianru;HUANG Meiying;XIONG Yalan;ZHENG Chenxi;WANG Zhigang;YUAN Gengbiao(Department of Nuclear Medicine,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010;Chongqing Key Laboratory of Ultrasound Molecular Imaging,Institute of Ultrasound Imaging,Chongqing Medical University,Chongqing,400016,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2020年第8期772-782,共11页
Journal of Third Military Medical University
基金
重庆市科技局基础研究与前沿探索(CSTC2018jcyjAX0747)
国家自然科学基金重点项目(31630026)。
关键词
酞菁铁
PLGA
适配体AS1411
乳腺癌
光声/超声成像
光热转化能力
iron(Ⅱ)phthalocyanine
poly(latic-co-glycolic acid)
aptamer AS1411
breast cancer
photoacoustic/ultrasonic imaging
photothermal conversion capacity
