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. 2022 Sep;53(5):752-757.
doi: 10.12182/20220960101.

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

[Article in Chinese]
Jian-Bo Huang  1 , Lin Yang  1 , Rui Yang  1 , Feng Yan  1 , Yu-Lan Peng  1 , Jin-Shun Xu  1
Affiliations

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

[Article in Chinese]
Jian-Bo Huang et al. Sichuan Da Xue Xue Bao Yi Xue Ban. 2022 Sep.

Abstract
in English, Chinese

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/cm 2. 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.

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。

Keywords: Enhanced ultrasound imaging; Fucoidan; Perfluorocarbons; Phase transition; Targeting.

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Conflict of interest statement

利益冲突 所有作者均声明不存在利益冲突

Figures

图 1
图 1
Physiochemical characterization of FPCA 制备的FPCA理化性质表征
图 2
图 2
Phase transition assay of FPCA FPCA相变实验
图 3
图 3
Ultrasound imaging assay of FPCA FPCA超声显像实验
图 4
图 4
Targeting assay of FPCA by laser confocal microscopy 激光共聚焦显微镜观察FPCA对细胞靶向性结果
图 5
图 5
Binding assay of FPCA and PCA in SMMC-7721 and L02 cell lines by flow cytometry 流式细胞术检测PCA和FPCA在两种细胞中的结合率
See this image and copyright information in PMC

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