Towards a synthetic basement membrane for the corneal epithelium

描述（由申请人提供）：对细胞功能有很大影响的基底膜特征包括顺应性、特异性蛋白质和官能团、生长因子和其他营养因子的储存库以及粘附细胞延伸突起和形成粘附斑的复杂三维形貌。三个三维亚微米和纳米级的底层基板的地形，基板的顺应性，独立于特定的受体-配体的相互作用，最近已被证明影响基本的细胞行为。该提案的重点是角膜上皮细胞与旨在整合到可植入设备中的仿生材料的相互作用。我们的设计参数将基于这样的前提，即模拟天然基底膜的结构、化学性质和顺应性将有利于宿主细胞对完全分化和功能性上皮的表面重塑和维持。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下基底膜的地形学模拟特征的相互作用，以确定用于改善组织结构和功能的设计参数，并整合到人工角膜的设计和制造中。在本申请中，提出了一种多学科的方法，使用定量形态学技术，细胞生物学，分子生物学和最先进的纳米级制造技术的体外方法来测试3菌丝。假设1：微米、亚微米和纳米级形貌和顺应性的组合将不同地影响细胞附着和扩散、增殖和迁移。假设二：结合肽序列和形貌的同质和异质呈现将协同影响细胞增殖、粘附和迁移。假设三：结合仿生地形，顺应性和表面化学将允许确定材料参数，以增强或延迟细胞行为在明确定义的区域内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.