Controlling Cell Phenotype in a Tissue-Engineered Corneal Model

控制组织工程角膜模型中的细胞表型

• 批准号: 8812691
• 负责人: Marta Keith Bechtel
• 金额: $ 39.15万
• 依托单位: HARVEY MUDD COLLEGE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8812691
• 关键词: Address; Affect; Animal Testing; Behavior; Biocompatible Materials; Biomimetics; Bioreactors; Cells; Chemicals; Clinical; Collagen; Cornea; Corneal Injury; Cues; Down-Regulation; Electromagnetics; Environment; Exposure to; Extracellular Matrix; Fibroblasts; Fibronectins; Gene Delivery; Genetic Transcription; Goals; Grant; Human; Immune; Integrins; Keratoplasty; Laser In Situ Keratomileusis; Lasers; Lead; Light; Link; Mechanics; Messenger RNA; Microscopy; Modeling; Myofibroblast; Optics; Outcome; Patients; Phenotype; Polymers; Population; Procedures; Proteins; Relative (related person); Research; Role; Sampling; Signal Transduction; Smooth Muscle Actin Staining Method; Stromal Cells; System; Time; Tissue Engineering; Tissues; Transplanted tissue; Visible Radiation; Vision; Western Blotting; Work; Wound Healing; aldehyde dehydrogenase 1; base; biomaterial compatibility; cell behavior; design; drug testing; improved; light scattering; novel; ophthalmic drug; partial recovery; protein expression; public health relevance; repaired; response; scaffold

DESCRIPTION (provided by applicant): A viable, transparent corneal model can provide the basis for studying novel ophthalmic drugs and new gene delivery approaches. Moreover, there is currently a clinical need for improved understanding of corneal wound-healing mechanisms in order to solve corneal haze problems associated with LASIK procedures and outcomes in corneal transplants. An understanding of the factors that contribute to the expression of the wound-healing phenotype in the cornea can lead to an understanding of how to control these changes. The overall goal of this proposed project is to understand and control the relationship between corneal cell behavior and resulting transparency in a tissue-engineered corneal model. A number of input signals to the cells have been characterized, including chemical, mechanical, electromagnetic, and topographical signals. The results of these studies taken together indicate that physical signals are more promising than chemical signals for the control of the wound-healing response in corneal fibroblasts. The proposed studies aim at understanding the differential, and perhaps more importantly, the synergistic effects of several signals in order to determine which signals or signal combinations are most successful as we move toward our ultimate goal of a three-dimensional corneal model. Cells will be analyzed for protein levels by western blot, mRNA levels will be analyzed by quantitative real-time PCR (qR-T PCR), and the extent of light scattering will be determined by optical coherence microscopy (OCM). Specifically, we will assess signaling associated with the wound healing response, integrin signaling, alpha-SMA, TKT, ALDH1, and matrix remodeling markers associated with changes in the keratocyte phenotype under the various treatment conditions detailed in this proposal. Cell behavior will be assessed to understand (1) the relative importance of matrix alignment and matrix composition; (2) the effect of matrix stiffness; and (3) the potentially synergistic effectsof light and mechanical strain input signals. By understanding the relative strengths of these input signals, we can more intelligently construct a transparent corneal matrix and model. Overall, these studies will lead to a better understanding of the role of biomimicry in the design and implementation of tissue-engineered systems. In addition, they will contribute to the understanding of wound healing in fibroblastic cells.

描述（由申请人提供）：可行的、透明的角膜模型可以为研究新型眼科药物和新的基因递送方法提供基础。此外，目前临床需要增进对角膜伤口愈合机制的了解，以解决与 LASIK 手术和角膜移植结果相关的角膜混浊问题。了解有助于角膜中伤口愈合表型表达的因素可以帮助我们了解如何控制这些变化。该项目的总体目标是了解和控制组织工程角膜模型中角膜细胞行为与透明度之间的关系。细胞的许多输入信号已经被表征，包括化学、机械、电磁和地形信号。这些研究的结果表明，在控制角膜成纤维细胞的伤口愈合反应方面，物理信号比化学信号更有希望。拟议的研究旨在了解几种信号的差异，也许更重要的是，了解几种信号的协同效应，以便确定哪些信号或信号组合在我们朝着三维角膜模型的最终目标迈进时最成功。将通过蛋白质印迹分析细胞的蛋白质水平，通过定量实时 PCR (qR-T PCR) 分析 mRNA 水平，并通过光学相干显微镜 (OCM) 确定光散射的程度。具体来说，我们将评估与伤口愈合反应、整合素信号、α-SMA、TKT、ALDH1 相关的信号传导，以及与本提案中详述的各种治疗条件下角膜细胞表型变化相关的基质重塑标记物。将评估细胞行为以了解（1）基质排列和基质组成的相对重要性； (2)基体刚度的影响； (3)光和机械应变输入信号的潜在协同效应。通过了解这些输入信号的相对强度，我们可以更智能地构建透明角膜矩阵和模型。总的来说，这些研究将有助于更好地理解仿生学在组织工程系统的设计和实施中的作用。此外，它们还将有助于理解成纤维细胞的伤口愈合。