Malakpour-Permlid Atena, Rodriguez Manuel Marcos, Untracht Gavrielle R, Andersen Peter E, Oredsson Stina, Boisen Anja, Zór Kinga
Center for Intelligent Drug Delivery and Sensing Using Microcontainers and Nanomechanics (IDUN), Department of Health Technology, Technical University of Denmark, Kongens Lyngby 2800, Denmark.
Department of Health Technology, Technical University of Denmark, Kongens Lyngby 2800, Denmark.
Toxicol Rep. 2024 Dec 12;14:101863. doi: 10.1016/j.toxrep.2024.101863. eCollection 2025 Jun.
High-throughput screening (HTS) three-dimensional (3D) tumor models are a promising approach for cancer drug discovery, as they more accurately replicate cell behavior than two-dimensional (2D) models. However, assessing and comparing current 3D models for drug efficacy remains essential, given the significant influence of cellular conditions on treatment response. To develop mimicking 3D models, we evaluated two HTS 3D models established in 96-well plates with 3D polycaprolactone (PCL) scaffolds fabricated using two distinct methods, resulting in scaffolds with either homogenous or non-homogenous fiber networks. These models, based on human HeLa cervical cancer cells and cancer-associated fibroblasts (CAFs) cultured as mono- or co-cultures within the 3D scaffolds, revealed that anticancer drug paclitaxel (PTX) exhibited consistently higher inhibitory concentration 50 (IC) in 3D (≥ 1000 nM) compared to 2D (≥ 100 nM), indicating reduced toxicity on cells cultured in 3D. Interestingly, the toxicity of PTX was significantly lower on mini-tumors in non-homogenous 3D (IC: 600 or 1000 nM) than in homogenous 3D cultures (IC exceeding 1000 nM). Microscopic studies revealed that the non-homogenous scaffolds closely resemble the tumor collagen network than their homogeneous counterpart. Both 3D scaffolds offer optimal pore size, facilitating efficient cell infiltration into the depth of 58.1 ± 1.2 µm (homogenous) and 86.4 ± 9.8 µm (non-homogenous) within 3D cultures. Cells cultured in the 3D non-homogenous systems exhibited drug treatment responses closer to conditions, highlighting the role of scaffold structure and design on cellular response to drug treatment. The PCL-based 3D models provide a robust, tunable, and efficient approach for the HTS of anti-cancer drugs compared to conventional 2D systems.
高通量筛选（HTS）三维（3D）肿瘤模型是癌症药物发现的一种有前景的方法，因为它们比二维（2D）模型更准确地复制细胞行为。然而，鉴于细胞条件对治疗反应的重大影响，评估和比较当前用于药物疗效的3D模型仍然至关重要。为了开发模拟3D模型，我们评估了两种在96孔板中建立的HTS 3D模型，这些模型使用两种不同方法制造的3D聚己内酯（PCL）支架，从而产生具有均匀或非均匀纤维网络的支架。这些基于人宫颈癌细胞系HeLa和癌症相关成纤维细胞（CAF）在3D支架内作为单培养或共培养的模型显示，与2D（≥100 nM）相比，抗癌药物紫杉醇（PTX）在3D（≥1000 nM）中始终表现出更高的半数抑制浓度（IC），表明对在3D中培养的细胞毒性降低。有趣的是，PTX对非均匀3D中的微型肿瘤（IC：600或1000 nM）的毒性明显低于均匀3D培养物（IC超过1000 nM）。显微镜研究表明，非均匀支架比均匀支架更类似于肿瘤胶原网络。两种3D支架都提供了最佳孔径，有利于细胞有效渗透到3D培养物中深度为58.1±1.2 µm（均匀）和86.4±9.8 µm（非均匀）的区域。在3D非均匀系统中培养的细胞表现出更接近体内条件的药物治疗反应，突出了支架结构和设计对细胞对药物治疗反应的作用。与传统的2D系统相比，基于PCL的3D模型为抗癌药物的高通量筛选提供了一种强大、可调节且高效的方法。
