Pamplona Regina, González-Lana Sandra, Romero Pilar, Ochoa Ignacio, Martín-Rapún Rafael, Sánchez-Somolinos Carlos
Aragón Institute of Nanoscience and Materials (INMA), CSIC-University of Zaragoza, Department of Organic Chemistry, C/ Pedro Cerbuna 12, Zaragoza, 50009, Spain.
BEONCHIP S.L., CEMINEM, Campus Río Ebro. C/ Mariano Esquillor Gómez s/n, Zaragoza, 50018, Spain.
Macromol Biosci. 2023 Dec;23(12):e2300227. doi: 10.1002/mabi.202300227. Epub 2023 Aug 24.
From the first experiments with biomaterials to mimic tissue properties, the mechanical and biochemical characterization has evolved extensively. Several properties can be described, however, what should be essential is to conduct a proper and physiologically relevant characterization. Herein, the influence of the reaction media (RM) and swelling media (SM)-phosphate buffered saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) with two different glucose concentrations-is described in gelatin methacrylamide (GelMA) hydrogel mechanics and in the biological behavior of two tumoral cell lines (Caco-2 and HCT-116). All scaffolds are UV-photocrosslinked under identical conditions and evaluated for mass swelling ratio and stiffness. The results indicate that stiffness is highly susceptible to the RM, but not to the SM. Additionally, PBS-prepared hydrogels exhibited a higher photopolymerization degree according to high resolution magic-angle spinning (HR-MAS) NMR. These findings correlate with the biological response of Caco-2 and HCT-116 cells seeded on the substrates, which demonstrated flatter morphologies on stiffer hydrogels. Overall, cell viability and proliferation are excellent for both cell lines, and Caco-2 cells displayed a characteristic apical-basal polarization based on F-actin/Nuclei fluorescence images. These characterization experiments highlight the importance of conducting mechanical testing of biomaterials in the same medium as cell culture.
从最初使用生物材料模拟组织特性的实验开始，力学和生化表征已经有了广泛的发展。可以描述多种特性，然而，至关重要的是要进行恰当且与生理相关的表征。在此，描述了反应介质（RM）和溶胀介质（SM）——含两种不同葡萄糖浓度的磷酸盐缓冲盐水（PBS）和杜氏改良 Eagle 培养基（DMEM）——对明胶甲基丙烯酰胺（GelMA）水凝胶力学性能以及两种肿瘤细胞系（Caco - 2 和 HCT - 116）生物学行为的影响。所有支架在相同条件下进行紫外光交联，并评估其质量溶胀率和刚度。结果表明，刚度对反应介质高度敏感，但对溶胀介质不敏感。此外，根据高分辨率魔角旋转（HR - MAS）核磁共振，用 PBS 制备的水凝胶表现出更高的光聚合度。这些发现与接种在基质上的 Caco - 2 和 HCT - 116 细胞的生物学反应相关，即在较硬的水凝胶上细胞形态更扁平。总体而言，两种细胞系的细胞活力和增殖情况都很好，并且基于 F - 肌动蛋白/细胞核荧光图像，Caco - 2 细胞呈现出典型的顶 - 基极化。这些表征实验突出了在与细胞培养相同的介质中对生物材料进行力学测试的重要性。
