Electrokinetic lithography: in situ microengineering of anisotropic 3D collagen matrices

摘要 在这个项目中，我们将开发一种称为电动光刻(EKL)的技术，它将填补一项技术 在目前最先进的3D排列胶原纤维微工程技术中处于空白状态。要模仿 在天然细胞外基质(ECM)中发现的结构不同的环境，我们将顺序 将伸展流体流动与电场驱动的微珠运动(电动)相结合，以“写”细胞尺度 仿生3D胶原凝胶中胶原纤维排列区域之间的不连续。技术 发展的目标如下：1)建立基于流动的胶原纤维排列和 表征电动传输参数，以及2)开发微流控平台来工程 在排列整齐的3D凝胶环境中的不连续性，并验证细胞运动响应。 该项目的成功将建立一个可转移的实验室原型，并支持前所未有的研究 探索细胞如何应对排列成行的纤维微结构中的局部破坏。我们的技术将 支持在结构上与移动性、传感和细胞间通信相关的新的探索路线 异类环境，并解决研究问题，目前还不能用 最先进的胶原蛋白排列技术。该项目直接与NIGMS开发工具的使命相一致 这使得潜在的变革性生物医学研究成为可能。

Summary In this project we will develop a technology called electrokinetic lithography (EKL) that will fill a technology void in the current state-of-the-art 3D aligned collagen fiber microengineering techniques. To mimic the structurally heterogenous environment found in the native extracellular matrix (ECM), we will sequentially combine extensional fluid flows with electric field driven bead motion (electrokinetics) to “write” cellular-scale discontinuities between domains of aligned collagen fibers within biomimetic 3D collagen gel. Technology development will be carried out with the followings aims: 1) Establish flow-based collagen fiber alignment and characterize electrokinetic transport parameters, and 2) Develop a microfluidic platform to engineer discontinuities within aligned 3D gel environments and validate cell motility responses. The success of this project will establish a transferrable lab prototype and support unprecedented studies that explore how cells respond to local disruptions in the aligned fibrous microarchitecture. Our technique will support new lines of exploration related to motility, sensing, and cell-cell communication within structurally heterogeneous environments, and address research questions cannot be currently answered with state-of- the-art collagen alignment techniques. This project directly aligns with the NIGMS mission of developing tools that enable potentially transformative biomedical research.