从最初的仿生材料实验到模拟组织特性，机械和生化特性已经得到了广泛的发展。有很多特性可以进行描述，然而，重要的是进行适当和具有生理相关性的表征。在此，我们描述了反应介质和缓冲介质（磷酸盐缓冲盐溶液（PBS）和Dulbecco修饰的Eagle培养基（DMEM）以两种不同葡萄糖浓度）对GelMA水凝胶力学特性和两种肿瘤细胞系（Caco-2和HCT-116）生物行为的影响。所有支架都是在相同条件下通过紫外线光固化，并进行了质量增重比和硬度评估。我们的结果表明，硬度对反应介质非常敏感，而对于溶胀介质则不敏感。此外，通过HR-MAS NMR确认，用PBS制备的水凝胶显示出更高的光聚合程度。这些发现与在基质上种植的Caco-2和HCT-116细胞的生物响应相一致，表明在更坚硬的水凝胶上，细胞呈现较平坦的形态。总的来说，两种细胞系的细胞存活和增殖都非常好，而Caco-2细胞在F-actin/核染色质荧光显微镜图像中显示了特征性的顶基极化。这些表征实验突出了在与细胞培养相同的介质中进行生物材料的力学测试的重要性。本文受版权保护。保留所有权利。

From the first experiments with biomaterials to mimic tissue properties, the mechanical and biochemical characterization have evolved extensively. A number of properties can be described, however, what should be essential is to conduct a proper and physiologically relevant characterization. Herein, we describe the influence of the reaction media and buffer media -phosphate buffer saline (PBS) and Dulbecco's modified Eagle's medium (DMEM) with two different glucose concentrations- in GelMA hydrogel mechanics and in the biological behavior of two tumoral cell lines (Caco-2 and HCT-116). All scaffolds were photocrosslinked using UV light under identical conditions and evaluated for mass swelling ratio and stiffness. Our results indicate that stiffness is highly susceptible to the reaction media, but not to the swelling media. In addition, PBS-prepared hydrogels exhibited a higher photopolymerization degree as confirmed by 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 were excellent for both cell lines, and Caco-2 cells displayed a characteristic apical-basal polarization as observed in F-actin/Nuclei fluorescence images. These characterization experiments highlight the importance of conducting mechanical testing of biomaterials in the same medium as cell culture. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.