Kim Hoyoung, Kim Eun-Ji, Ngo Hai V, Nguyen Hy D, Park Chulhun, Choi Kyung Hyun, Park Jun-Bom, Lee Beom-Jin
College of Pharmacy, Ajou University, Suwon 16499, Republic of Korea.
College of Pharmacy, Jeju National University, Jeju 63243, Republic of Korea.
Pharmaceuticals (Basel). 2023 Sep 20;16(9):1330. doi: 10.3390/ph16091330.
The objective of this study was to evaluate the effectiveness of organ-on-chip system investigating simultaneous cellular efficacy and real-time reactive oxygen species (ROS) occurrence of anticancer drug-loaded nanoparticles (NPs) using hepatocarcinoma cells (HepG2) chip system under static and hepatomimicking shear stress conditions (5 dyne/cm). Then, the role of hepatomimetic shear stress exposed to HepG2 and drug solubility were compared. The highly soluble doxorubicin (DOX) and poorly soluble paclitaxel (PTX) were chosen. Fattigated NPs (AONs) were formed via self-assembly of amphiphilic albumin (HSA)-oleic acid conjugate (AOC). Then, drug-loaded AONs (DOX-AON or PTX-AON) were exposed to a serum-free HepG2 medium at 37 °C and 5% carbon dioxide for 24 h using a real-time ROS sensor chip-based microfluidic system. The cellular efficacy and simultaneous ROS occurrence of free drugs and drug-loaded AONs were compared. The cellular efficacy of drug-loaded AONs varied in a dose-dependent manner and were consistently correlated with real-time of ROS occurrence. Drug-loaded AONs increased the intracellular fluorescence intensity and decreased the cellular efficacy compared to free drugs under dynamic conditions. The half-maximal inhibitory concentration (IC) values of free DOX (13.4 μg/mL) and PTX (54.44 μg/mL) under static conditions decreased to 11.79 and 38.43 μg/mL, respectively, under dynamic conditions. Furthermore, DOX- and PTX-AONs showed highly decreased IC values of 5.613 and 21.86 μg/mL, respectively, as compared to free drugs under dynamic conditions. It was evident that cellular efficacy and real-time ROS occurrence were well-correlated and highly dependent on the drug-loaded nanostructure, drug solubility and physiological shear stress.
本研究的目的是评估芯片器官系统的有效性，该系统使用肝癌细胞（HepG2）芯片系统，在静态和模拟肝剪切应力条件（5达因/厘米）下，研究载抗癌药物纳米颗粒（NPs）的细胞疗效和实时活性氧（ROS）生成情况。然后，比较了模拟肝剪切应力对HepG2细胞的作用以及药物溶解度。选择了高溶解性的阿霉素（DOX）和低溶解性的紫杉醇（PTX）。通过两亲性白蛋白（HSA）-油酸共轭物（AOC）的自组装形成脂肪化纳米颗粒（AONs）。然后，使用基于实时ROS传感器芯片的微流控系统，将载药AONs（DOX-AON或PTX-AON）在37℃、5%二氧化碳条件下于无血清HepG2培养基中孵育24小时。比较了游离药物和载药AONs的细胞疗效及同时的ROS生成情况。载药AONs的细胞疗效呈剂量依赖性变化，且与ROS的实时生成始终相关。在动态条件下，与游离药物相比，载药AONs增加了细胞内荧光强度并降低了细胞疗效。静态条件下，游离DOX（13.4μg/mL）和PTX（54.44μg/mL）的半数抑制浓度（IC）值在动态条件下分别降至11.79和38.43μg/mL。此外，与动态条件下的游离药物相比，DOX-AONs和PTX-AONs的IC值分别大幅降至5.613和21.86μg/mL。显然，细胞疗效与实时ROS生成密切相关，且高度依赖于载药纳米结构、药物溶解度和生理剪切应力。
