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Easy Manipulation of Architectures in Protein-based Hydrogels for Cell Culture Applications

机译:轻松操纵基于蛋白质的水凝胶在细胞培养应用中的架构

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摘要

Hydrogels are recognized as promising materials for cell culture applications due to their ability to provide highly hydrated cell environments. The field of 3D templates is rising due to the potential resemblance of those materials to the natural extracellular matrix. Protein-based hydrogels are particularly promising because they can easily be functionalized and can achieve defined structures with adjustable physicochemical properties. However, the production of macroporous 3D templates for cell culture applications using natural materials is often limited by their weaker mechanical properties compared to those of synthetic materials. Here, different methods were evaluated to produce macroporous bovine serum albumin (BSA)-based hydrogel systems, with adjustable pore sizes in the range of 10 to 70 µm in radius. Furthermore, a method to generate channels in this protein-based material that are several hundred microns long was established. The different methods to produce pores, as well as the influence of pore size on material properties such as swelling ratio, pH, temperature stability, and enzymatic degradation behavior, were analyzed. Pore sizes were investigated in the native, swollen state of the hydrogels using confocal laser scanning microscopy. The feasibility for cell culture applications was evaluated using a cell-adhesive RGD peptide modification of the protein system and two model cell lines: human breast cancer cells (A549) and adenocarcinomic human alveolar basal epithelial cells (MCF7).
机译:由于水凝胶具有提供高度水合的细胞环境的能力,因此被认为是用于细胞培养的有前途的材料。由于这些材料与天然细胞外基质的潜在相似性,因此3D模板的领域正在增加。基于蛋白质的水凝胶特别有前途,因为它们可以轻松地功能化并可以实现具有可调节理化性质的确定结构。但是,与天然材料相比,使用天然材料生产用于细胞培养的大孔3D模板通常受到其较弱的机械性能的限制。在这里,评估了不同的方法来生产基于大孔牛血清白蛋白(BSA)的水凝胶系统,其孔径可调范围为半径10到70 µm。此外,建立了一种在这种基于蛋白质的材料中产生数百微米长的通道的方法。分析了产生孔的不同方法,以及孔径对材料特性(如溶胀率,pH,温度稳定性和酶促降解行为)的影响。使用共聚焦激光扫描显微镜研究了水凝胶天然,膨胀状态下的孔径。使用蛋白质系统的细胞粘附性RGD肽修饰和两种模型细胞系(人乳腺癌细胞(A549)和腺癌人类肺泡基底上皮细胞(MCF7))评估了细胞培养应用的可行性。

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