In vivo, cells interact with the extracellular matrix (ECM), which provides a multitude of biophysical and biochemical signals that modulate cellular behavior. Inspired by this, we explored a new methodology to develop a more physiomimetic polysaccharide-based matrix for 3D cell culture. Maleimide-modified alginate (AlgM) derivatives were successfully synthesized using DMTMM to activate carboxylic groups. Thiol-terminated cell-adhesion peptides were tethered to the hydrogel network to promote integrin binding. Rapid and efficient hydrogel formation was promoted by thiol-Michael addition “click” chemistry via maleimide reaction with thiol-flanked protease-sensitive peptides. Alginate derivatives were further ionically crosslinked by divalent ions present in the medium, which led to greater stability and allowed longer cell culture periods. By tailoring alginate's biofunctionality we improved cell-cell and cell-matrix interactions, providing an ECM-like 3D microenvironment. We were able to systematically and independently vary biochemical and biophysical parameters to elicit specific cell responses, creating custom-made 3D matrices. DMTMM-mediated maleimide incorporation is a promising approach to synthesizing AlgM derivatives that can be leveraged to produce ECM-like matrices for a broad range of applications, from tissue modeling to tissue regeneration.

中文翻译：

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通过硫醇-马来酰亚胺点击化学原位交联多功能和细胞响应性藻酸盐 3D 基质

在体内，细胞与细胞外基质 (ECM) 相互作用，细胞外基质提供大量调节细胞行为的生物物理和生化信号。受此启发，我们探索了一种新方法来开发用于 3D 细胞培养的更加拟态的多糖基质。使用DMTMM激活羧基成功合成了马来酰亚胺改性海藻酸盐（AlgM）衍生物。硫醇封端的细胞粘附肽被束缚在水凝胶网络上以促进整合素结合。通过马来酰亚胺与硫醇侧翼蛋白酶敏感肽的反应，硫醇-迈克尔加成“点击”化学促进了快速有效的水凝胶形成。海藻酸盐衍生物通过培养基中存在的二价离子进一步离子交联，这导致更高的稳定性并允许更长的细胞培养时间。通过定制海藻酸盐的生物功能，我们改善了细胞-细胞和细胞-基质的相互作用，提供了类似 ECM 的 3D 微环境。我们能够系统地、独立地改变生化和生物物理参数，以引发特定的细胞反应，从而创建定制的 3D 矩阵。 DMTMM 介导的马来酰亚胺掺入是合成 AlgM 衍生物的一种有前途的方法，可用于生产类似 ECM 的基质，用于从组织建模到组织再生的广泛应用。