载锰卟啉/紫杉醇的相变纳米粒多模态成像及靶向治疗肿瘤的体外实验研究  

In vitro experimental study of drug-MnTTP/paclitaxel nanoparticles with phase-changed capability for tumor multimodal imaging and sonodynamic therapy combined with chemotherapy

作　　者：张奕[1] 罗远利 刘当 曹阳[2] 冉海涛[2] 杨超 ZHANG Yi;LUO Yuanli;LIU Dang;CAO Yang;RAN Haitao;YANG Chao(Department of Ultrasound,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第一医院超声科,重庆市400010 [2]重庆医科大学附属第二医院超声科 [3]重庆医科大学生物医学工程学院 [4]中国科学院大学重庆医院,重庆市人民医院放射科

出　　处：《临床超声医学杂志》2021年第10期721-726,共6页Journal of Clinical Ultrasound in Medicine

基　　金：国家自然科学基金面上项目(82071926);国家自然科学基金重点项目(81630047);重庆市人民医院医学科技创新项目(2019ZDXM08)。

摘　　要：目的制备载全氟戊烷/锰卟啉/紫杉醇的聚乳酸-羟基乙醇酸(PLGA)纳米粒(PFP-MnTTP-PTX@PLGA,以下简称FMP@P),评估其体外光声、超声、磁共振多模态成像及其与声动力治疗(SDT)联合化疗肿瘤的效果。方法采用双乳化法制备FMP@P,应用透射电镜、扫描电镜观察纳米粒的形态,马尔文粒径电位分析仪检测其粒径及表面电位,紫外-可见光分光光度计检测其吸光度并计算包封率;高效液相法测量并计算MnTTP-PTX和紫杉醇的包封率及超声辐照后的药物释放能力。采集不同浓度下纳米粒的体外光声、磁共振成像图像,定量对比分析其显像效果;使用低功率聚焦超声(3.0 W/cm2)辐照纳米粒,采集不同辐照时间的超声成像图像,定量对比分析其显像效果。使用单线态氧荧光探针检测纳米粒的活性氧产生能力。通过CCK-8法及流式细胞术验证FMP@P与SDT联合化疗杀伤肿瘤细胞的效果。结果FMP@P乳液为墨绿色,纳米粒为均一的球形,平均粒径为(323.1±68.3)nm,平均表面电位为(-14.5±8.3)mV,MnTTP-PTX和紫杉醇的包封率分别为(93.55±0.46)%和(81.23±6.93)%。药物释放实验结果显示FMP@P在超声辐照后24 h PTX的累计释放率为31.57%。FMP@P可以增强超声B模式和造影模式的显像能力,回声强度随辐照时间的增加而增强;也可增强光声、磁共振显像能力,光声信号、磁共振信号强度随FMP@P浓度的增加而增强。超声辐照后乳腺癌4T1细胞内产生大量活性氧。FMP@P与SDT联合化疗可显著杀伤肿瘤细胞。结论本实验成功制备了FMP@P,其不但可体外增强光声、超声、磁共振显像能力,还可介导SDT联合化疗有效治疗肿瘤。Objective To prepare polylactic-co-glycolic acid(PLGA)based multifunctional nanoparticles loading with perfluoropentane(PFP),MnTTP and paclitaxel(PTX),termed as FMP@P,and to investigate its ultrasound(US)/photoacoustic(PA)/magnetic resonance(MR)imaging capability and chemo-/sonodynamic therapy(SDT)efficacy in vitro.Methods FMP@P was prepared by a double emulsification method.The morphology of nanoparticles was observed by transmission electron microscope and scanning electron microscope.The particle size and surface potential were detected by Malvern particle size potentiometer.The absorbance was detected by UV-visible spectrophotometer and the encapsulation efficiency was calculated.The encapsulation efficiency of PTX and the drug release ability after US irradiation were determined by high performance liquid chromatography method.In addition,the in vitro PA and MR images of nanoparticles at different concentrations were collected for quantitative comparison and analysis.The nanoparticles were irradiated by low intensity focused ultrasound(3.0 W/cm2),and the US images were collected at different times for quantitative comparison and analysis.The reactive oxygen production capacity of nanoparticles was detected by singlet oxygen fluorescence probe.Meanwhile,the chemo-/sonodynamic therapy effect on tumor cells was evaluated by the standard CCK-8 assay and flow cytometry.Results The FMP@P emulsion was dark green,the nanoparticles were uniformly spherical,the average diameter and Zeta potential of FMP@P were(323.1±68.3)nm and(-14.5±8.3)mV,respectively.The encapsulation efficiency of MnTTP-PTX and PTX were(93.55±0.46)%and(81.23±6.93)%,respectively.FMP@P could enhance the capability of US/PA/MR imaging,and the intensity was increased with the concentration increased.After US irradiation,FMP@P could produce a large amount of reactive oxygen species in 4T1 cell.The combined of FMP@P with SDT and chemotherapy can significantly kill tumor cells.Conclusion This experiment successfully prepared FMP@P,which can not only

关 键 词：纳米粒 光声成像 超声成像 磁共振成像 声动力治疗 化疗 

分 类 号：R445.1[医药卫生—影像医学与核医学] R73-36[医药卫生—诊断学]