Biomolecular Patterning of 3-Dimensional Polymeric Microscaffolds for Targeted Cell Attachment

来自软物质合成和细胞生物学领域的两个小组联手开发用于三维直接激光写入（DLW）聚合物微支架的生物功能化的精密光触发化学。一个目的是将这些图案化的功能支架应用于上皮细胞和内皮细胞的附着，以探索诱导顶基底极性的可能性。此外，将测量成纤维细胞施加的细胞力与粘附几何形状、粘附位点的数量和大小、细胞外基质（ECM）的类型或呈递给相同细胞的细胞粘附分子的关系。该项目分为两个独立但相互交织的工作方案：高分子化学导向的工作计划旨在建立在我们的初步工作领域，采用光触发化学在空间分辨的方式，生物分子表面固定，通过严格扩大可用的光触发化学的反应类型，从而波长制度，用于附件（在DLW后官能化或原位DLW官能化过程方面）以及通过提供通过光诱导的可逆化学使细胞附着解除缀合的方法。此外，该项目的聚合物化学部分将为特定功能化支架以及可变刚度的DLW书面支架提供有效的钝化方法。生物驱动的工作计划将利用已开发的化学工具，通过构建细胞信号分子库，这些分子可以连接到DLW写入的光反应支架上。本文开发的方法将提供一个基础，系统地研究三维的组合影响，粘附/信号蛋白的空间分布，以及在单细胞水平上的细胞行为的环境力学。

Two groups from the respective fields of soft matter synthesis and cell biology join forces to develop precision light triggered chemistries for the bio- functionalization of three dimensional direct laser written (DLW) polymer microscaffolds. One aim is to apply these patterned functional scaffolds for the attachment of epithelial and endothelial cells in order to probe the possibility of inducing apicobasal polarity. In addition, cellular forces applied by fibroblasts will be measured in relation to the adhesion geometry, the number and size of adhesion sites, the type of extracellular matrix (ECM) or cell adhesion molecules presented to the same cell. The project is divided into two separate, yet intertwined work programs: The polymer chemistry oriented work program seeks to built on our preliminary work in the field of employing light triggered chemistry in a spatially resolved fashion for biomolecule surface immobilization by critically expanding the available light triggered chemistries in terms of the reaction type and thus wavelength regime employed for the attachment (either in terms of post-DLW functionalization or in situ DLW functionalization processes) as well as by providing approaches to unconjugate cell attachments by light induced reversible chemistries. In addition, the polymer chemistry part of the project will provide access to efficient passivation approaches for the site specifically functionalized scaffolds as well as to DLW written scaffolds of variable stiffness. The biologically driven work program will capitalize on the developed chemical tools by building a library of cell signaling molecules that can be tethered to the DLW written photoreactive scaffolds. The herein developed methods will provide a basis to systematically study the combined impacts of three-dimensionality, of the spatial distribution of adhesion/signaling proteins, and of environmental mechanics on cell behavior at the single cellular level.