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

黄建波; 杨林; 阳锐; 阎锋; 彭玉兰; 徐金顺

doi:10.12182/20220960101

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

摘要

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/cm²。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。

Abstract:
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 phase-transition 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/cm². Moreover, after phase transition, FPCA showed significant imaging enhancement on both two-dimensional ultrasonography and contrast-enhanced 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.

HTML全文

参考文献(21)

施引文献

资源附件(0)

开放获取 本文遵循知识共享署名—非商业性使用4.0国际许可协议（CC BY-NC 4.0），允许第三方对本刊发表的论文自由共享（即在任何媒介以任何形式复制、发行原文）、演绎（即修改、转换或以原文为基础进行创作），必须给出适当的署名，提供指向本文许可协议的链接，同时标明是否对原文作了修改；不得将本文用于商业目的。CC BY-NC 4.0许可协议详情请访问 https://creativecommons.org/licenses/by-nc/4.0