Towards a synthetic basement membrane for the corneal epithelium

描述（由申请人提供）：基底膜的特征极大地影响细胞功能，包括顺应性，特定蛋白质和功能基团，生长因子和其他营养因子的储存库，以及粘附细胞延伸过程并形成粘附斑块的复杂三维地形。三维亚微米和纳米尺度的底物形貌和底物的顺应性，独立于特定的受体-配体相互作用，最近被证明影响基本的细胞行为。这一建议的重点是角膜上皮细胞与仿生材料的相互作用，设计集成到一个植入式装置。我们的设计参数将基于模拟天然基底膜的结构、化学和顺应性的前提，这将有助于宿主细胞重新表面和维持完全分化和功能的上皮。本提案的总体目的是确定顺应性、通过粘附肽配体的表面化学和角膜上皮基底膜的地形模拟特征之间的相互作用，以确定改善组织结构和功能的设计参数，并将其整合到角膜假体的设计和制造中。在本应用中，提出了一种多学科的方法来测试3个假设，使用定量形态学技术，细胞生物学，分子生物学的体外方法和最先进的纳米级制造技术。假设1：结合微观、亚微米和纳米尺度的地形和顺应性会不同地影响细胞的附着、扩散、增殖和迁移。假设2：结合多肽序列和地形的同质和异质表现将协同影响细胞增殖、粘附和迁移。假设3：结合仿生地形、顺应性和表面化学，将允许确定材料参数，以增强或延缓明确区域内的细胞行为，以整合到新的角膜等效物中。

DESCRIPTION (provided by applicant): Basement membrane features that greatly influence cell function include compliance, specific proteins and functional groups, a reservoir of growth factor and other trophic agents, and a complex three-dimensional topography into which adherent cells extend processes, and formadhesion plaques. Thre three-dimensional submicron and nanoscale topography of the underlying substrate, and substrate compliance, independent of specific receptor-ligand interactions has been recently shown to influence fundamental cell behaviors. This proposal focuses on the interactions of corneal epithelial cells with biomimetic materials designed for integration into an implantable device. Our design parameters will be based on the premise that mimicking the structure, chemistry and compliance of the native basement membrane will be conducive to resurfacing and maintenance of a fully differentiated and functional epithelium by host cells. The overall purpose of this proposal is to determine the interaction of compliance, surface chemistry via adhesive peptide ligands, and topography mimicking features of the basement membrane underlying the corneal epithelium to determine design parameters for improved tissue architecture and function, and for integration into the design and fabrication of keratoprostheses. In this application, a multi-disciplinary approach is proposed to test 3 hyphothese using quantitative morphologic techniques, in vitro methodologies in cell biology, molecular biology and state-of-the-art nanoscale fabrication techniques. Hypothesis 1: Combined micro, submicron, and nanoscale topography and compliance will differentially affect cell attachment and spreading, proliferation, and migration. Hypothesis 2: Combining homogeneous and heterogeneous presentations of peptide sequences and topography will synergistically impact cell proliferation, adhesion and migration. Hypothesis 3: Combining biomimetic topography, compliance and surface chemistry will allow for the determination of materials parameters to enhance or retard cell behaviors within well-defined areas for integration into novel corneal equivalents.