岩藻多糖修饰的相变型造影剂体外超声显像及靶向肝癌的实验研究  

Fucoidan-Modified Phase-Transitional Contrast Agent for Ultrasound Imaging and Targeting of Hepatoma Cells

作　　者：黄建波 杨林[1] 阳锐 阎锋 彭玉兰[1] 徐金顺 HUANG Jian-bo;YANG Lin;YANG Rui;YAN Feng;PENG Yu-lan;XU Jin-shun(Department&Institute of Ultrasound Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院超声医学科/超声医学研究室,成都610041

出　　处：《四川大学学报（医学版）》2022年第5期752-757,共6页Journal of Sichuan University(Medical Sciences)

基　　金：北京市自然科学基金(No.7192200);成都市科技局项目(No.2019-YF05-00376-SN);四川省卫生健康委员会项目(No.20PJ011)资助。

摘　　要：目的制备岩藻多糖修饰的相变型造影剂(FPCA),评价其体外超声显影和对肝癌细胞的靶向能力。方法采用薄膜水化-超声乳化法制备包裹全氟戊烷的纳米脂质体,之后化学接枝岩藻多糖,制备FPCA纳米粒,对其基本理化性质进行表征。使用显微镜观察FPCA在水浴加热、微波辐照后的相变情况;使用超声诊断仪观察FPCA相变后的二维超声、超声造影成像能力;使用荧光共聚焦、流式细胞仪分析验证FPCA对肝癌细胞的靶向能力。结果制备的FPCA平均粒径(222.1±32.5)nm,外观呈球形,分散性好,具有较好的稳定性和生物相容性。使用水浴加热、微波辐照均能使FPCA发生相变,最佳相变温度为50℃,最佳相变微波功率为1.5 W/cm^(2)。并且,相变后的FPCA对二维超声、超声造影均具有显著的增强显像能力。通过使用FITC荧光标记,FPCA能够特异地与肝癌细胞靶向结合,结合率高达(96.19±1.62)%,而对正常肝脏细胞的结合率不足10%。结论成功制备出稳定性和生物相容性良好的新型相变超声造影剂,其不仅可通过相变增强超声显像,还能对肝癌细胞具有特异的主动靶向性能。Objective To prepare a fucoidan-modified phase-transitional contrast agent(FPCA)and to evaluate its in vitro capabilities for ultrasound imaging and targeting of hepatoma cells.Methods Nano-liposomes encapsulated with perfluoropentane were prepared using thin-film hydration and ultrasonic emulsification methods.Then,FPCA nanoparticles were prepared through chemical grafting of fucoidan and the characterization of their physical and chemical properties was performed.After applying external stimuli of heating with hot water bath and microwave irradiation,the phase-transition status of FPCA was observed with microscope.The imaging abilities of phase-transited FPCA on two-dimensional ultrasound and contrast-enhanced ultrasound were observed with ultrasonic diagnostic instrument.The ability of FPCA to target at hepatoma cells was evaluated and verified with fluorescence confocal observation and flow cytometry analysis.Results The FPCA prepared in the study had an average diameter of(222.1±32.5)nm,displaying spherical appearance,good dispersion,good stability,and good biocompatibility.The phasetransition of FPCA was induced by both heating with hot water bath and microwave irradiation.For phase transition,the optimal temperature was found to be 50℃and the preferred microwave power was 1.5 W/cm^(2).Moreover,after phase transition,FPCA showed significant imaging enhancement on both two-dimensional ultrasonography and contrastenhanced ultrasonography.Through fluorescein isothiocyanate(FITC)labeling,FPCA could specifically bind with hepatoma cells at a high binding rate of(96.19±1.62)%,while it rarely bound with normal liver cells,showing a binding rate of less than 10%.Conclusion A new type of phase-transitional ultrasound contrast agent with good stability and biocompatibility was successfully prepared.It not only could enhance ultrasound imaging through phase transition,but also had specific active hepatoma cell-targeting properties.

关 键 词：全氟化碳 岩藻多糖 相变 靶向性 增强显像 

分 类 号：R445.1[医药卫生—影像医学与核医学] R735.7[医药卫生—诊断学]