Characterizing cell adhesion and migration of surfaces biofunctionalized with nanopatterned collagen matrices

我们最近开发了一种方法，用于在非生物表面上创建纳米（~ 3 nm）胶原基质，其中单个胶原微纤维在纳米级上几乎完全有序。初步实验表明，通过调整胶原蛋白基质的纳米形貌特征，可以精确控制不同细胞类型的粘附和迁移特性。未来使用这种纳米图案化的胶原蛋白基质作为表面生物功能化的智能可编程涂层，需要深入了解细胞对图案化基质反应的分子机制。含有整合素的粘附复合物，如粘着斑，是细胞-基质粘附的主要介质，并将细胞骨架连接到细胞外基质。因此，为了理解我们的纳米图案化基质的微纤维如何调节细胞形态和迁移，阐明它们如何指导粘着斑的形成和调节细胞骨架重排将是至关重要的。我们打算调查的影响，纳米图案化的胶原蛋白晶格上的粘着斑形成和细胞骨架重排在分子尺度上采用光和原子力显微镜（AFM）的方法相结合。此外，胶原纳米形貌对整合素介导的粘附的影响将使用AFM单分子力光谱进行定量。阐明细胞对胶原蛋白基质反应的分子机制不仅有助于改善其生物相容性，促进智能生物功能材料的设计，而且还将为细胞识别和响应周围地形特征的机制提供新的见解。

We have recently developed a method for creating ultrathin (~ 3 nm) collagen matrices on non-biological surfaces in which individual collagen microfibrils are almost perfectly ordered on the nanoscale. Initial experiments show that by adjusting nanotopographic features of the collagen matrix, the adhesive and migratory properties of different cell types can be precisely controlled. The future use of such nanopatterned collagen matrices as intelligent, programmable coatings for the biofunctionalization of surfaces requires a profound understanding of the molecular mechanisms underlying the cellular response to the patterned matrix. Integrin-containing adhesion complexes, such as focal adhesions, are the main mediators of cell-substratum adhesion and link the cytoskeleton to the extracellular matrix. Consequently, for understanding how the microfibrils of our nanopatterned matrices regulate cell morphology and migration it will be crucial to elucidate how they direct the formation of focal adhesions and regulate cytoskeletal rearrangement. We intend to investigate the influence of nanopatterned collagen lattices on focal adhesion formation and cytoskeletal rearrangement at a molecular scale by employing a combination of light and atomic force microscopy (AFM) methods. Furthermore, the effect of the collagen nanotopography on integrin-mediated adhesion will be quantitated using AFM single molecule force spectroscopy. Elucidation of the molecular mechanisms underlying the cellular response to collagen matrices will not only help to improve their biocompatibility and facilitate the design of intelligent, biofunctionalized materials, but it will also offer new insight into the mechanisms by which cells recognize and respond to topographic features in their surrounding.