Lens Epithelial Cell Response to Biomaterial Interfaces

晶状体上皮细胞对生物材料界面的反应

• 批准号: 10372517
• 负责人: Katelyn E Swindle-Reilly
• 金额: $ 18.26万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372517
• 关键词: Affect; Anterior; Apoptosis; Biocompatible Materials; Biological; Biology; Biomedical Engineering; Blindness; Cataract; Cataract Extraction; Cell physiology; Cells; Clinical; Complication; Conflict (Psychology); Crystalline Lens; Development; Disease; E-Cadherin; Epithelial Cell Proliferation; Epithelial Cells; Excision; Extracellular Matrix; Eye; Fibronectins; Fibrosis; Formulation; Gene Expression; Gene Proteins; Glass; Goals; Hydrogels; Immunofluorescence Immunologic; Implant; In Vitro; Incidence; Intraocular Lens Implantation; Intraocular lens implant device; Investigation; Knowledge; Lens Fiber; Link; Mesenchymal; Methods; Microspheres; Modeling; Operative Surgical Procedures; Pathologic; Phenotype; Polymers; Procedures; Process; Production; Property; Publications; Radial; Reporting; Research; Research Personnel; Residual state; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Shapes; Signaling Protein; Sirolimus; Snails; Stress; Surface; Surface Properties; Tissues; Transforming Growth Factor beta; United States; Vision; Western Blotting; Work; base; biomaterial interface; capsule; cell behavior; cell motility; copolymer; design; epithelial to mesenchymal transition; experimental study; implantation; improved; innovation; lens; lens capsule; mechanical properties; migration; physical property; prevent; protein expression; response; slug; tool; viscoelasticity

PROJECT SUMMARY Cataract remains the leading cause of blindness worldwide with over 3 million extractions performed each year in the United States alone. During cataract surgery, the contents inside the lens capsule are removed through a hole in the anterior lens capsule and a polymeric intraocular lens (IOL) is placed in the capsule. The leading vision-threatening complication, posterior capsule opacification (PCO), occurs when residual lens epithelial cells (LEC) migrate from the anterior to the posterior lens capsule or onto the IOL and undergo epithelial-to- mesenchymal transition (EMT). While several factors impacting mechanobiology and epithelial cell response have been previously investigated, there is not a clear understanding of the impact of viscoelasticity and curvature on LEC behavior. The overall objective of this project is to use polymer and hydrogel substrates that mimic the implants and lens microenvironment, respectively, to better analyze the influence mechanical properties have on LEC response and EMT. It is hypothesized that the physical and mechanical properties of the microenvironment are altered after the removal of the lens tissue and IOL placement, facilitating EMT in LEC. In Aim 1, tunable polymer substrates and hydrogels will be used to investigate the impact of stiffness and viscoelasticity on LEC response and EMT. It is hypothesized that substrates stiffer than the lens capsule, and substrates with lower loss tangent will drive EMT in LEC. In Aim 2, the effect of substrate curvature will be investigated using the same polymer and hydrogel substrates. The governing hypothesis is that LEC migration and EMT are driven by larger radius of curvature caused by flattening of the lens capsule after IOL implantation. Curvature effects will be evaluated using polymers micropatterned with different radii of curvature. Glass microbeads of various sizes will be embedded in hydrogel formulations, mimicking the changes in the lens capsule shape following surgery. In both aims, relevant in vitro and ex vivo models will be used. LEC proliferation, migration, and markers for EMT will be assessed. TGF-β and rapamycin will be used as positive and negative inducers of EMT, respectively. RT-PCR will quantify gene expression, and changes in protein expression will be evaluated using Western blot and immunofluorescence. Specific genes and proteins that will be evaluated include SMAD signaling proteins, α-SMA, Slug, Snail, fibronectin, E-cadherin, and YAP. The goal of this project is to determine how substrate mechanical properties, namely viscoelasticity and curvature, contribute to LEC behavior and induction of EMT. This will significantly enhance our knowledge of LEC mechanobiology and the role of these factors in EMT, suggesting strategies to prevent pathological EMT. The results will lead to future research on design of materials to prevent EMT and fibrosis after implantation, particularly for preventing PCO.

项目概要 白内障仍然是全球失明的主要原因，每年进行超过 300 万例摘除手术 仅在美国，在白内障手术期间，晶状体囊内的内容物通过手术被取出。 晶状体前囊上有孔，囊内有聚合物人工晶状体 (IOL)。 当晶状体上皮细胞残留时，会发生威胁视力的并发症，即后囊混浊（PCO） (LEC) 从晶状体前囊迁移到晶状体后囊或迁移到 IOL 上，并经历上皮-到- 间质转化（EMT），同时影响机械生物学和上皮细胞反应的几个因素。 之前已经进行过研究，但对于粘弹性和粘弹性的影响还没有清楚的认识 该项目的总体目标是使用聚合物和水凝胶基底 分别模拟植入物和晶状体微环境，以更好地分析机械影响 特性对 LEC 响应和 EMT 的物理和机械特性有影响。 去除晶状体组织和放置 IOL 后微环境发生改变，促进 LEC 中的 EMT。 在目标 1 中，可调谐聚合物基底和水凝胶将用于研究刚度和 粘弹性对 LEC 响应和 EMT 的影响被重新认识到基底比晶状体囊更硬，并且 具有较低损耗角正切的基板将驱动 LEC 中的 EMT 在目标 2 中，基板曲率的影响将是 使用相同的聚合物和水凝胶基质进行了研究，主导假设是 LEC 迁移。 EMT是由IOL植入后晶状体囊变平导致的较大曲率半径驱动的。 将使用具有不同曲率半径的微图案聚合物玻璃来评估曲率效应。 各种尺寸的微珠将嵌入水凝胶配方中，模仿晶状体的变化 手术后的胶囊形状将使用相关的体外和离体模型。 迁移，EMT 标记物将被用作阳性和阴性评估。 EMT 诱导剂将分别量化基因表达和蛋白质表达的变化。 使用蛋白质印迹和免疫荧光进行评估 将评估的特定基因和蛋白质。 包括 SMAD 信号蛋白、α-SMA、Slug、Snail、纤连蛋白、E-钙粘蛋白和 YAP。 是为了确定基材机械特性（即粘弹性和曲率）如何影响 LEC EMT 的行为和诱导这将显着增强我们对 LEC 机械生物学和 这些因素在EMT中的作用，提出了预防病理性EMT的策略，结果将导致未来。 研究防止植入后EMT和纤维化的材料设计，特别是预防PCO。