摘要
目的构建一种癌细胞膜包覆的仑伐替尼(LEN)仿生纳米颗粒(L-P-S-CM),旨在改善药物溶解性并增强同源靶向递送潜力,深入探究其pH响应释放动力学。方法采用复乳液溶剂蒸发与物理挤出技术制备L-P-S-CM,表征其粒径与Zeta电位,并测定载药量和包封率;在模拟生理(pH7.4)与肿瘤微环境(pH5.5)下进行体外释放实验,并利用零级、一级、Higuchi及Korsmeyer-Peppas等动力学模型对数据进行拟合分析。基于游离仑伐替尼的剂量-效应曲线拟合计算其IC_(50),并据此选取等效仑伐替尼浓度为2,4μg/mL开展细胞毒性评价。在上述浓度条件下,将人肝癌细胞Huh-7分为游离LEN组以及不同纳米制剂(PLGA、L-P、P-S、L-P-S、L-P-S-CM)组,正常人肝细胞THLE-2分为PLGA组、L-P-S组和L-P-S-CM组,采用CCK-8法测定各组细胞存活率并进行比较。结果L-P-S-CM包封率高达93.33%,在酸性条件(pH5.5)下释药快于pH7.4(12 h:42.71%vs 17.10%;72 h:76.40%vs 55.64%),呈现pH依赖性释放。在零级、一级、Higuchi及Korsmeyer-Peppas等模型中比较后,一级动力学模型的拟合优度最高(决定系数R^(2)≥0.982)。Korsmeyer-Peppas模型分析揭示,细胞膜包覆在pH7.4下将释放机制从Fickian扩散(n≈0.43)转变为非Fickian扩散(n=0.567),提示其引入了骨架松弛效应。该膜层延缓了药物突释,并在酸性环境中表现出一定缓冲作用。细胞实验显示,游离仑伐替尼对Huh-7细胞的IC_(50)为1.6652μg/mL;在等效仑伐替尼浓度为4μg/mL时,L-P-S-CM组Huh-7与THLE-2细胞存活率分别为(16.91±0.22)%和(82.09±0.21)%。结论成功构建癌细胞膜包覆的仑伐替尼仿生纳米颗粒L-P-S-CM,呈pH响应性释药特征(酸性条件下释药更快);在等效仑伐替尼浓度条件下L-P-S-CM对Huh-7细胞具有抑制作用,同时THLE-2细胞保持较高存活率。
Objective To develop a cancer cell membrane-coated biomimetic nanoparticle(L-P-S-CM)for lenvatinib(LEN)delivery,aiming to improve its solubility,enhance the potential for homotypic targeted delivery,and investigate its pH-responsive release kinetics.Methods L-P-S-CM was fabricated via a double emulsion solvent evaporation method coupled with physical extrusion.The nanopar⁃ticles were comprehensively characterized for size,zeta potential,drug loading(DL),and encapsulation efficiency(EE).In vitro release profiles were evaluated under simulated physiological(pH7.4)and tumor microenvironment(pH5.5)conditions.The release data were fitted using various kinetic models,including zero-order,first-order,Higuchi,and Korsmeyer-Peppas models.Based on the dose-response curve of free LEN,the IC_(50) for Huh-7 cells was calculated.Accordingly,LEN-equivalent concentrations(2,4μg/mL)were chosen for cytotoxicity evaluation.Huh-7 hepatocellular carcinoma cells were treated with free LEN and nanoparticle formulations(PLGA,L-P,P-S,L-P-S,and L-P-S-CM),whereas THLE-2 normal human liver cells were treated with PLGA,L-P-S,and L-P-S-CM,at the selected LEN-equivalent concentrations.Cell viability was measured using the cell counting kit-8(CCK-8)assay.Results L-P-SCM achieved a high encapsulation efficiency(93.33%)and exhibited faster drug release at pH 5.5 than at pH7.4(12 h:42.71%vs 17.10%;72 h:76.40%vs 55.64%),demonstrating pH-dependent release.Among the compared kinetic models,the first-order model showed the best goodness of fit(R^(2)≥0.982).Korsmeyer-Peppas model analysis indicated that the cell membrane coating shifted the release mechanism from Fickian diffusion(n≈0.43)to non-Fickian diffusion(n=0.567)at pH7.4,suggesting the introduction of a polymer matrix relaxation effect.The membrane layer delayed the initial burst release and exhibited a buffering effect under acidic conditions.Cytotoxicity assays showed that the half-maximal inhibitory concentration(IC_(50))of free LEN for Huh-7 cells was 1.6652μg/mL.At a LEN-equivalent concentration of 4μg/mL,the viability of Huh-7 and THLE-2 cells in the L-P-S-CM group was(16.91±0.22)%and(82.09±0.21)%,respectively.Conclusion L-P-S-CM is successfully constructed and exhibits pH-responsive release(faster release under acidic conditions).At LEN-equivalent concentrations,L-P-S-CM can reduce Huh-7 cell viability,at the same time,THLE-2 cells maintain relatively high viability.
作者
武思学
沈建松
温艳艳
田伟
WU Sixue;SHEN Jiansong;WEN Yanyan;TIAN Wei(Academy of Medical Sciences,Shanxi Medical University,Taiyuan 030001,China;Department of General Surgery,Cardiovascular Hospital Affiliated to Shanxi Medical University;NHC Key Laboratory of Pneumoconiosis,Shanxi Key Laboratory of Respiratory Diseases,Department of Pulmonary and Critical Care Medicine,The First Hospital of Shanxi Medical University;Department of General Surgery,Shanxi Cardiovascular Hospital)
出处
《山西医科大学学报》
2026年第2期185-192,共8页
Journal of Shanxi Medical University
基金
山西省卫生健康委科研课题计划项目(2023108)
山西省心血管病医院科研激励计划项目(XYS20220109)。
关键词
仑伐替尼
仿生纳米颗粒
pH响应释放
癌细胞膜
药物释放动力学
Lenvatinib
biomimetic nanoparticles
pH-responsive release
cancer cell membrane
drug release kinetics
