Towards a synthetic basement membrane for the corneal epithelium

描述(申请人提供)：显著影响细胞功能的基底膜特征包括顺应性、特定的蛋白质和功能基团、生长因子和其他营养物质的储存库，以及贴壁细胞延伸过程和形成黏附斑块的复杂三维地形。最近研究表明，底物的三维亚微米和纳米尺度的形貌和底物的顺应性，与特定的受体-配体相互作用无关，影响基本的细胞行为。这项提议的重点是角膜上皮细胞与仿生材料之间的相互作用，这些材料被设计成集成到可植入设备中。我们的设计参数将基于这样的前提，即模仿天然基底膜的结构、化学成分和顺应性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细胞。这项建议的总体目的是确定顺应性、通过粘附性多肽配体的表面化学以及模仿角膜上皮下基底膜的地形图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，并整合到角膜假体的设计和制造中。在这一应用中，提出了一种多学科的方法来测试3种菌丝体，使用定量形态技术、体外细胞生物学方法、分子生物学和最新的纳米级制造技术。假设1：结合微米、亚微米和纳米尺度的地形和顺应性将不同地影响细胞的附着和扩散、增殖和迁移。假设2：将多肽序列的同质和异质呈现与拓扑学相结合，将协同影响细胞的增殖、黏附和迁移。假设3：结合仿生地形学、顺应性和表面化学，将允许确定材料参数，以增强或抑制定义明确的区域内的细胞行为，从而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.