Collagen hydrogels with controllable combined cues of elasticity and topography to regulate cellular processes
Collagen hydrogels with controllable combined cues of elasticity and topography to regulate cellular processes
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DOI：
10.1088/1748-605x/ac0452
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发表时间：
2021-07-01
影响因子：
4
通讯作者：
Taguchi, Mitsumasa
中科院分区：
文献类型：
--
作者：
Oyama, Tomoko G.;Oyama, Kotaro;Taguchi, Mitsumasa
细胞培养基质的弹性、形貌和化学成分影响细胞行为。然而，细胞对体内细胞外基质（ECM）（蛋白质（主要是胶原蛋白）和多糖的水凝胶）的反应仍然未知，因为没有保留天然ECM关键特征的底物。本研究介绍了新型胶原蛋白水凝胶，其可以结合联合收割机弹性、形貌和组成，并再现天然ECM中胶原蛋白浓度（C）和弹性模量（E）之间的相关性。一种简单的无试剂方法，其基于辐射交联改变的ECM衍生的胶原I和水解的胶原（明胶或胶原肽）溶液形成水凝胶，所述水凝胶具有可调的弹性模量，所述弹性模量覆盖广泛范围的软组织（E = 1-236 kPa），所述弹性模量源自水凝胶中的最终胶原密度（C = 0.3%-14%）和精确的微观形貌（> 1 μ m）。该方法制备的胶原蛋白水凝胶的氨基酸组成比例基本不变，且保持了胶原蛋白的酶介导降解性。这些胶原蛋白水凝胶能够研究细胞系（成纤维细胞、上皮细胞和成肌细胞）和原代细胞（大鼠心肌细胞）对软地形线索的反应，例如在C和E之间正相关的体内反应。这些细胞直接粘附在胶原蛋白水凝胶上，并根据E（即胶原蛋白网络的密度）和C选择停留在顶部或自发迁移到其中。我们发现，细胞形态和肌动蛋白细胞骨架组织符合地形线索，即使当他们在体内ECM一样软。胶原蛋白水凝胶上更硬的微槽更有效地排列细胞，除了HeLa细胞在细胞形态上发生了剧烈变化。这些胶原蛋白水凝胶不仅可以减少体内和体外细胞行为差异，而且还可以促进人工ECM设计以控制细胞功能和命运，用于组织工程和再生医学。
The elasticity, topography, and chemical composition of cell culture substrates influence cell behavior. However, the cellular responses to in vivo extracellular matrix (ECM), a hydrogel of proteins (mainly collagen) and polysaccharides, remain unknown as there is no substrate that preserves the key features of native ECM. This study introduces novel collagen hydrogels that can combine elasticity, topography, and composition and reproduce the correlation between collagen concentration (C) and elastic modulus (E) in native ECM. A simple reagent-free method based on radiation-cross-linking altered ECM-derived collagen I and hydrolyzed collagen (gelatin or collagen peptide) solutions into hydrogels with tunable elastic moduli covering a broad range of soft tissues (E = 1-236 kPa) originating from the final collagen density in the hydrogels (C = 0.3%-14%) and precise microtopographies (> 1 mu m). The amino acid composition ratio was almost unchanged by this method, and the obtained collagen hydrogels maintained enzyme-mediated degradability. These collagen hydrogels enabled investigation of the responses of cell lines (fibroblasts, epithelial cells, and myoblasts) and primary cells (rat cardiomyocytes) to soft topographic cues such as those in vivo under the positive correlation between C and E. These cells adhered directly to the collagen hydrogels and chose to stay atop or spontaneously migrate into them depending on E, that is, the density of the collagen network, C. We revealed that the cell morphology and actin cytoskeleton organization conformed to the topographic cues, even when they are as soft as in vivo ECM. The stiffer microgrooves on collagen hydrogels aligned cells more effectively, except HeLa cells that underwent drastic changes in cell morphology. These collagen hydrogels may not only reduce in vivo and in vitro cell behavioral disparity but also facilitate artificial ECM design to control cell function and fate for applications in tissue engineering and regenerative medicine.