One-Step Synthesis of Gelatin-Conjugated Supramolecular Hydrogels for Dynamic Regulation of Adhesion Contact and Morphology of Myoblasts
One-Step Synthesis of Gelatin-Conjugated Supramolecular Hydrogels for Dynamic Regulation of Adhesion Contact and Morphology of Myoblasts
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DOI：
10.1021/acsapm.1c01902
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发表时间：
2022-04-08
影响因子：
5
通讯作者：
Tanaka, Motomu
中科院分区：
文献类型：
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作者：
Hayashi, Kentaro;Matsuda, Mami;Tanaka, Motomu
水凝胶具有精细可调节和可切换的弹性，模拟生物细胞的机械微环境，这些环境在发育和疾病进展过程中是动态变化的。本研究合成了一种具有明胶侧链的超分子水凝胶。通过系统筛选超分子主/客体交联剂的摩尔分数，底物的杨氏模量被微调到成肌细胞的水平，E接近10 kPa。C2C12成肌细胞通过由整合素团簇组成的黏附接触与明胶共轭水凝胶具有可重复性和牢固的黏附，而只有少数细胞与不含明胶侧链的凝胶有黏附。通过简单地添加和去除适当浓度的游离客体分子，而不干扰细胞活力，明胶共轭水凝胶的弹性可以切换到所需的水平。固定细胞的免疫荧光共聚焦显微镜图像证实了明胶共轭水凝胶上的局灶粘连和肌动蛋白细胞骨架的重塑。延时相衬图像显示了细胞的动态响应，表现在它们的形态上，对衬底弹性的突然变化。具有可切换弹性的明胶共轭水凝胶能够在一步中直接和可逆地对细胞进行机械刺激，而无需使用粘附配体进行繁琐的表面功能化。
Hydrogels possessing fine-adjustable and switchable elasticity emulate the mechanical microenvironments of biological cells, which are known to change dynamically during development and disease progression. In this study, a supramolecular hydrogel conjugated with gelatin side chains was synthesized. By systematically screening the molar fraction of supramolecular host/guest cross-linkers, Young's modulus of the substrate was fine-adjusted to the level for myoblasts, E approximate to 10 kPa. C2C12 myoblasts reproducibly and firmly adhered to the gelatin-conjugated hydrogel via focal adhesion contacts consisting of integrin clusters, whereas only a few cells adhered to the gel without gelatin side chains. The elasticity of the gelatin-conjugated hydrogel was switchable to desired levels by simply adding and removing free guest molecules in appropriate concentrations without interfering with cell viability. Immunofluorescence confocal microscopy images of fixed cells confirmed the adaptation of focal adhesions and remodeling of actin cytoskeletons on the gelatinconjugated hydrogel. Time-lapse phase-contrast images demonstrated the dynamic response of the cells, manifested in their morphology, to an abrupt change in the substrate elasticity. Gelatin-conjugated hydrogels with switchable elasticity enable the direct and reversible mechanical stimulation of cells in one step without tedious surface functionalization with adhesion ligands.