基于上转换纳米粒子多糖探针的构建及其在肿瘤成像中的应用

doi:10.6040/j.issn.1671-7554.0.2024.0430

摘要/Abstract

摘要： 目的构建一种用上转换纳米颗粒(upconversion nanoparticles, UCNPs)与硫酸软骨素(chondroitin sulfate, CS)连接的探针,开发肝细胞癌(hepatocellular carcinoma, HCC)早期诊断的方法。方法以己二酸二酰肼(adipic acid hydrazide, ADH)为桥梁,构建UCNPs-CS探针。首先,CS在1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐［1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, EDC］和 N-羟基琥珀酰亚胺(N-hydroxysuccinimide, NHS)的催化下合成CS-ADH,CS具有肿瘤靶向性,可作为探针的靶向基团。然后,由溶剂热法制备NaYF4:Yb,Tm@NaYF4核壳结构的UCNPs,制备的UCNPs的荧光发射带在800 nm左右,可作为荧光报告基团用于体内成像。UCNPs通过聚丙烯酸(polyacrylic acid, PAA)衍生转水后由羧基氨基偶联反应制备UCNPs-CS纳米荧光探针。UCNPs经光谱、核磁、透射电镜等对形貌和光学性质进行表征,并通过MTT实验对探针的细胞毒性进行考察。通过构建肝癌小鼠模型,评价探针体内成像和肿瘤靶向能力,并利用体内成像评估不同抗肿瘤药物的活性。结果制备的核壳结构UCNPs形貌均匀,大小均一,尺寸约35 nm,壳层厚度约3~4 nm,在水中分散性好。构建的UCNPs-CS探针具有良好的发光性能(激发波长980 nm,荧光发射波长800 nm)和细胞相容性(在质量浓度高达500 μg/mL时,24 h后的细胞存活率大于60%),具有较强的组织穿透力,可实现动物水平的活体成像。肝癌小鼠体内成像结果表明,构建的UCNPs-CS探针具有较好的肿瘤靶向性,且成像光辐射强度与肿瘤体积呈正相关。结论构建的UCNPs-CS探针为HCC早期可视化诊断提供一种新工具,同时也为抗肿瘤药物的活性筛选提供一种新思路。

关键词: 硫酸软骨素, 上转换纳米颗粒, 肝细胞癌, 生物成像, 药物活性筛选

Abstract: Objective To develop a probe for early diagnosis of hepatocellular carcinoma(HCC)by connecting upconversion nanoparticles(UCNPs)with chondroitin sulfate(CS). Methods The UCNPs-CS nanoprobe was constructed using adipic acid hydrazide(ADH)as a bridging agent. First, CS, which exhibits tumor-targeting properties and serves as the targeting moiety of the probe, was functionalized to synthesize CS-ADH under the catalysis of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(EDC)and N-hydroxysuccinimide(NHS). Subsequently, NaYF4:Yb,Tm@NaYF4 UCNPs with a core-shell structure were prepared via a solvothermal method. The synthesized UCNPs exhibited fluorescence emission bands around 800 nm, making them suitable as fluorescent reporters for in vivo imaging. The UCNPs were rendered water-dispersible through polyacrylic acid(PAA)surface modification and then conjugated with CS-ADH via carboxyl-amine coupling to form the UCNPs-CS nanoprobe. The morphology, optical properties, and chemical composition of the UCNPs were characterized using spectroscopy, nuclear magnetic resonance(NMR), transmission electron microscopy(TEM), and other analytical techniques. The cytotoxicity of the probe was evaluated via MTT assay. To assess the probes in vivo imaging performance and tumor-targeting efficacy, hepatoma-bearing mouse models were established. Furthermore, the activity of different antitumor drugs was evaluated using in vivo imaging. Results The prepared UCNPs with core-shell structures had uniform morphology and size, with a size of about 35 nm and a shell thickness of about 3-4 nm. They had good dispersibility in water. The constructed UCNPs-CS probe had good luminescence performance(excitation wavelength 980 nm, fluorescence emission wavelength 800 nm), cell compatibility(cell survival rate greater than 60% after 24 hours at a concentration of up to 500 μg/mL), strong tissue penetration ability, and could achieve in vivo imaging at the animal level. The imaging results of liver cancer mice showed that the constructed UCNPs-CS probe had good tumor targeting ability, and the imaging radiation intensity was positively correlated with tumor volume. Conclusion The UCNPs-CS probe provides a new tool for early visual diagnosis of HCC, and also provides a new idea for the activity screening of anti-tumor drugs.

Key words: Chondroitin sulfate, Upconversion nanoparticles, Hepatocellular carcinoma, In vivo imaging, Drug activity screening

中图分类号:

R917

巩洁,于淼,李秀勇,陈颖,徐倩茹,李梅娟,李艺彤,刘秀美. 基于上转换纳米粒子多糖探针的构建及其在肿瘤成像中的应用[J]. 山东大学学报 (医学版), 2025, 63(11): 18-26.

GONG Jie, YU Miao, LI Xiuyong, CHEN Ying, XU Qianru, LI Meijuan, LI Yitong, LIU Xiumei. Construction of polysaccharide probe based on upconversion nanoparticles and its application in tumor bioimaging[J]. Journal of Shandong University (Health Sciences), 2025, 63(11): 18-26.

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