细胞骨架支架的主动与被动交联可调节细胞内含物对复合结构的影响
Active and passive crosslinking of cytoskeleton scaffolds tune the effects of cell inclusions on composite structure
AI总结：
该研究设计了嵌入细菌细胞的肌动蛋白-微管复合动态支架，验证了0.4%-2%体积分数细胞的嵌入可行性，揭示了主被动交联对细胞裹挟的促进作用及低细胞占比下的结构演化规律，为细胞调控的自主材料设计提供了参考。
摘要：
将细胞整合到活性生物材料支架中，是开发能够自主感知、响应环境信号或细胞内部回路并改变支架特性的前沿材料的一种极具前景的策略。采用可通过交联和马达驱动的力产生过程自行改变性能的动态生物相容性支架，为材料的驱动与调控开辟了更多可能性。然而，与嵌入细胞的活性支架工程化相关的设计原理尚未得到充分确立。为解决这一挑战，我们设计了一种动态支架材料：将细菌细胞嵌入由肌动蛋白和微管构成的复合细胞骨架网络中，该网络可通过生物素-链霉亲和素（被动交联）或多聚驱动蛋白马达（主动交联）实现被动或主动交联。借助定量显微镜技术，我们证明了能够在整个网络中嵌入体积分数为0.4%至2%的细胞，且不会破坏网络的结构完整性，也不会抑制交联过程或马达驱动的动力学行为。研究结果表明，被动交联和主动交联均会促进细胞被网络裹挟，而耗尽相互作用在未交联网络中发挥着更重要的作用。此外，我们发现即使细胞体积分数低至0.4%，也会出现宏观尺度的结构，但这些结构并不会影响材料的微观结构长度尺度。本工作凸显了我们的复合生物材料在设计由细胞调控的自主材料方面的潜力，并为有效将细胞与复杂复合材料耦合提供了路线图，旨在将细胞用作原位工厂来编程材料改性。
英文摘要：
Incorporating cells within active biomaterial scaffolds is a promising strategy to develop forefront materials that can autonomously sense, respond, and alter the scaffold in response to environmental cues or internal cell circuitry. Using dynamic biocompatible scaffolds that can self-alter their properties via crosslinking and motor-driven force-generation opens even greater avenues for actuation and control. However, the design principles associated with engineering active scaffolds embedded with cells are not well established. To address this challenge, we design a dynamic scaffold material of bacteria cells embedded within a composite cytoskeletal network of actin and microtubules that can be passively or actively crosslinked by either biotin-streptavidin or multimeric kinesin motors. Using quantitative microscopy, we demonstrate the ability to embed cells of volume fractions 0.4 to 2% throughout the network without compromising the structural integrity of the network or inhibiting crosslinking or motor-driven dynamics. Our findings suggest that both passive and active crosslinking promote entrainment of cells within the network, while depletion interactions play a more important role in uncrosslinked networks. Moreover, we show that large-scale structures emerge with the addition of cell fractions as low as 0.4%, but these structures do not influence the microscale structural lengthscale of the materials. Our work highlights the potential of our composite biomaterial in designing autonomous materials controlled by cells, and provides a roadmap for effectively coupling cells to complex composite materials with an eye towards using cells as in situ factories to program material modifications.
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