Zhou Yang, Pereira Gillian, Tang Yuanzhang, James Matthew, Zhang Miqin
Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA.
Department of Neurological Surgery, University of Washington, Seattle, WA 98195, USA.
Pharmaceutics. 2023 Jun 9;15(6):1691. doi: 10.3390/pharmaceutics15061691.
Natural polymer-based porous scaffolds have been investigated to serve as three-dimensional (3D) tumor models for drug screening owing to their structural properties with better resemblance to human tumor microenvironments than two-dimensional (2D) cell cultures. In this study, a 3D chitosan-hyaluronic acid (CHA) composite porous scaffold with tunable pore size (60, 120 and 180 µm) was produced by freeze-drying and fabricated into a 96-array platform for high-throughput screening (HTS) of cancer therapeutics. We adopted a self-designed rapid dispensing system to handle the highly viscous CHA polymer mixture and achieved a fast and cost-effective large-batch production of the 3D HTS platform. In addition, the adjustable pore size of the scaffold can accommodate cancer cells from different sources to better mimic the in vivo malignancy. Three human glioblastoma multiforme (GBM) cell lines were tested on the scaffolds to reveal the influence of pore size on cell growth kinetics, tumor spheroid morphology, gene expression and dose-dependent drug response. Our results showed that the three GBM cell lines showed different trends of drug resistance on CHA scaffolds of varying pore size, which reflects the intertumoral heterogeneity across patients in clinical practice. Our results also demonstrated the necessity to have a tunable 3D porous scaffold for adapting the heterogeneous tumor to generate the optimal HTS outcomes. It was also found that CHA scaffolds can produce a uniform cellular response (CV < 0.15) and a wide drug screening window (Z' > 0.5) on par with commercialized tissue culture plates, and therefore, can serve as a qualified HTS platform. This CHA scaffold-based HTS platform may provide an improved alternative to traditional 2D-cell-based HTS for future cancer study and novel drug discovery.
基于天然聚合物的多孔支架已被研究用作药物筛选的三维（3D）肿瘤模型，因为其结构特性比二维（2D）细胞培养更类似于人类肿瘤微环境。在本研究中，通过冷冻干燥制备了具有可调孔径（60、120和180 µm）的3D壳聚糖-透明质酸（CHA）复合多孔支架，并将其制成用于癌症治疗药物高通量筛选（HTS）的96孔阵列平台。我们采用了自行设计的快速分配系统来处理高粘性的CHA聚合物混合物，并实现了3D HTS平台的快速且经济高效的大批量生产。此外，支架的可调孔径可以容纳来自不同来源的癌细胞，以更好地模拟体内的恶性肿瘤。在支架上测试了三种多形性胶质母细胞瘤（GBM）细胞系，以揭示孔径对细胞生长动力学、肿瘤球体形态、基因表达和剂量依赖性药物反应的影响。我们的结果表明，三种GBM细胞系在不同孔径的CHA支架上表现出不同的耐药趋势，这反映了临床实践中患者间的肿瘤间异质性。我们的结果还证明了需要有一个可调的3D多孔支架来适应异质性肿瘤，以产生最佳的HTS结果。还发现CHA支架可以产生与商业化组织培养板相当的均匀细胞反应（CV < 0.15）和宽药物筛选窗口（Z' > 0.5），因此可以作为一个合格的HTS平台。这种基于CHA支架的HTS平台可能为未来癌症研究和新药发现提供一种优于传统基于2D细胞的HTS的改进替代方案。
