Isert Lorenz, Passi Mehak, Freystetter Benedikt, Grab Maximilian, Roidl Andreas, Müller Christoph, Mehta Aditi, Sundararaghavan Harini G, Zahler Stefan, Merkel Olivia M
Pharmaceutical Technology and Biopharmaceutics, Department of Pharmacy, Ludwig-Maximilians-University München, Munich, Germany.
Pharmaceutical Biology, Department of Pharmacy, Ludwig-Maximilians-Universität München, Munich, Germany.
Mater Today Bio. 2024 Dec 10;30:101401. doi: 10.1016/j.mtbio.2024.101401. eCollection 2025 Feb.
In this study, an advanced nanofiber breast cancer model was developed and systematically characterized including physico-chemical, cell-biological and biophysical parameters. Using electrospinning, the architecture of tumor-associated collagen signatures (TACS5 and TACS6) was mimicked. By employing a rotating cylinder or static plate collector set-up, aligned fibers (TACS5-like structures) and randomly orientated fibers (TACS6-like structures) fibers were produced, respectively. The biocompatibility of these fibers was enhanced by collagen coating, ensuring minimal toxicity and improved cell attachment. Various breast cancer cell lines (MCF7, HCC1954, MDA-MB-468, and MDA-MB-231) were cultured on these fibers to assess epithelial-to-mesenchymal transition (EMT) markers, cellular morphology, and migration. Aligned fibers (TACS5) significantly influenced EMT-related changes, promoting cellular alignment, spindle-shaped morphology and a highly migratory phenotype in mesenchymal and hybrid EMT cells (MDA-MB-468, MDA-MB-231). Conversely, epithelial cells (MCF7, HCC1954) showed limited response, but - under growth factor treatment - started to infiltrate the fibrous scaffold and underwent EMT-like changes, particularly on TACS5-mimicks, emphasizing the interplay of topographical cues and EMT induction. The biophysical analysis revealed a clear correlation between cellular EMT status and cell mechanics, with increased EMT correlating to decreased total cellular stiffness. Cancer cell mechanics, however, were found to be dynamic during biochemical and topographical EMT-induction, exceeding initial stiffness by up to 2-fold. These findings highlight the potential of TACS5-like nanofiber scaffolds in modeling the tumor microenvironment and studying cancer cell behavior and mechanics.
在本研究中，开发了一种先进的纳米纤维乳腺癌模型，并对其物理化学、细胞生物学和生物物理参数进行了系统表征。利用静电纺丝技术，模拟了肿瘤相关胶原特征（TACS5和TACS6）的结构。通过采用旋转圆柱或静态平板收集器设置，分别制备了排列的纤维（TACS5样结构）和随机取向的纤维（TACS6样结构）。通过胶原涂层提高了这些纤维的生物相容性，确保了最小的毒性并改善了细胞附着。在这些纤维上培养了各种乳腺癌细胞系（MCF7、HCC1954、MDA-MB-468和MDA-MB-231），以评估上皮-间质转化（EMT）标志物、细胞形态和迁移。排列的纤维（TACS5）显著影响与EMT相关的变化，促进间充质和混合EMT细胞（MDA-MB-468、MDA-MB-231）中的细胞排列、纺锤形形态和高度迁移表型。相反，上皮细胞（MCF7、HCC1954）显示出有限的反应，但在生长因子处理下开始浸润纤维支架并经历类似EMT的变化，特别是在TACS5模拟物上，强调了地形线索与EMT诱导之间的相互作用。生物物理分析揭示了细胞EMT状态与细胞力学之间的明显相关性，EMT增加与总细胞硬度降低相关。然而，发现癌细胞力学在生化和地形EMT诱导过程中是动态的，比初始硬度增加了2倍。这些发现突出了TACS5样纳米纤维支架在模拟肿瘤微环境以及研究癌细胞行为和力学方面的潜力。
