Many essential cellular processes are regulated by mechanical properties of their microenvironment. Here, we introduce stimuli-responsive composite scaffolds fabricated by three-dimensional (3D) laser lithography to simultaneously stretch large numbers of single cells in tailored 3D microenvironments. The key material is a stimuli-responsive photoresist containing cross-links formed by noncovalent, directional interactions between β-cyclodextrin (host) and adamantane (guest). This allows reversible actuation under physiological conditions by application of soluble competitive guests. Cells adhering in these scaffolds build up initial traction forces of ~80 nN. After application of an equibiaxial stretch of up to 25%, cells remodel their actin cytoskeleton, double their traction forces, and equilibrate at a new dynamic set point within 30 min. When the stretch is released, traction forces gradually decrease until the initial set point is retrieved. Pharmacological inhibition or knockout of nonmuscle myosin 2A prevents these adjustments, suggesting that cellular tensional homeostasis strongly depends on functional myosin motors.

许多重要的细胞过程受其微环境的机械特性的调节。在这里，我们介绍了通过三维 (3D) 激光光刻制造的刺激响应复合支架，以在定制的 3D 微环境中同时拉伸大量单细胞。关键材料是一种刺激响应光刻胶，它含有由 β-环糊精（主体）和金刚烷（客体）之间的非共价定向相互作用形成的交联。这允许通过应用可溶性竞争客体在生理条件下进行可逆驱动。粘附在这些支架中的细胞建立了约 80 nN 的初始牵引力。在应用高达 25% 的等双轴拉伸后，细胞会重塑其肌动蛋白细胞骨架，使其牵引力加倍，并在 30 分钟内平衡到新的动态设定点。当拉伸被释放时，牵引力逐渐减小，直到恢复初始设定点。非肌肉肌球蛋白 2A 的药理抑制或敲除可防止这些调整，这表明细胞张力稳态强烈依赖于功能性肌球蛋白马达。