DataSheet3_Physical properties and cellular responses of gelatin methacryloyl bulk hydrogels and highly ordered porous hydrogels.pdf

Protein-based hydrogels hold a high content of water in their three-dimensional (3D) network structure and exhibit innate biological activities as well as soft tissue-like mechanical properties, resulting in being highly applicable to various tissue engineering fields. However, precisely controlling the 3D porous structure of protein-based hydrogels remains a challenging task, and understanding the influence of their porous structure on physical properties and cellular responses is crucial for tissue engineering applications. In this study, we prepared highly ordered gelatin methacryloyl hydrogels with regular interconnected pores and traditional bulk hydrogels with irregular pores to evaluate their differences in physiochemical properties and cellular behaviors. Highly ordered gelatin methacryloyl hydrogels exhibited a high degree of compliance owing to their sponge-like structure whereas gelatin methacryloyl bulk hydrogels exhibited relatively higher moduli but were brittle due to a densely packed structure. The highly ordered gelatin methacryloyl hydrogels with interconnected pores supported higher cell viability (about 100%) due to an efficient flux of oxygen and nutrients compared to the dense bulk hydrogels showing cell viability (around 80%). Also, cells in the highly ordered gelatin methacryloyl hydrogels displayed a more stretched morphology compared to those in the gelatin methacryloyl bulk hydrogels that exhibited a more round morphology during the cell culture period.

基于蛋白质的水凝胶（Protein-based hydrogels）在三维（3D）网状结构中可容纳大量水分，兼具固有生物活性与类软组织的力学特性，因此在各类组织工程领域具备极高应用潜力。然而，精准调控基于蛋白质的水凝胶的三维多孔结构仍是一项极具挑战的任务，而阐明其多孔结构对物理特性与细胞响应的影响，对于组织工程应用而言至关重要。本研究制备了具备规则连通孔隙的高度有序甲基丙烯酸酯化明胶（gelatin methacryloyl）水凝胶，以及孔隙不规则的传统块状水凝胶，以对比二者在理化特性与细胞行为上的差异。具备规则连通孔隙的高度有序甲基丙烯酸酯化明胶水凝胶因类海绵结构展现出极高的柔顺性，而甲基丙烯酸酯化明胶块状水凝胶虽模量相对更高，但因结构致密堆砌而表现出脆性。相较于孔隙致密的块状水凝胶（细胞存活率约80%），具备连通孔隙的高度有序甲基丙烯酸酯化明胶水凝胶可维持更高的细胞存活率（约100%），这得益于其能高效输送氧气与营养物质。此外，在细胞培养周期内，接种于高度有序甲基丙烯酸酯化明胶水凝胶的细胞呈现出更为舒展的形态，而接种于甲基丙烯酸酯化明胶块状水凝胶的细胞则表现出更圆润的形态。