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: 10544163
• 负责人: Katelyn E Swindle-Reilly
• 金额: $ 22.11万
• 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/10544163
• 关键词: Affect; Anterior; Apoptosis; Biocompatible Materials; Biological Factors; Biology; Biomedical Engineering; Blindness; Cataract; Cataract Extraction; Cell physiology; Cells; Chemistry; Clinical; Complication; Crystalline Lens; Development; Disease; E-Cadherin; Epithelial Cell Proliferation; Epithelial Cells; Excision; Extracellular Matrix; Eye; Fibronectins; Fibrosis; Formulation; Gene Expression; Genes; 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; Pattern; Phenotype; Polymers; Procedures; Process; Production; Property; Proteins; 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; Viscosity; Vision; Western Blotting; Work; biomaterial interface; capsule; cell behavior; cell motility; copolymer; design; epithelial to mesenchymal transition; experimental study; implantation; improved; innovation; lens; lens capsule; lens induction; 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.

项目摘要 白内障仍然是全球失明的主要原因，每年进行300万次提取 仅在美国。在白内障手术期间，透镜胶囊内部的内容通过 将前晶状体囊和聚合物内透镜（IOL）的孔放在胶囊中。领导 当残留晶状体上皮细胞时，发生威胁性威胁并发症，后囊胶体拼布（PCO） （LEC）从前镜头或IOL迁移到IOL，并经历上皮到上皮 间充质转变（EMT）。而几个影响机制和上皮细胞反应的因素 以前已经研究过，对粘弹性的影响和 关于LEC行为的曲率。该项目的总体目的是使用聚合物和水凝胶底物 分别模仿即兴和晶状体微环境，以更好地分析影响机械的影响 属性具有LEC响应和EMT。假设的是 去除透镜组织和IOL放置后，微环境会改变，并在LEC中支撑EMT。 在AIM 1中，可调聚合物底物和水凝胶将用于研究刚度和 LEC响应和EMT的粘弹性。假设底物比镜头胶囊更硬，并且 损失切线较低的底物将驱动LEC中的EMT。在AIM 2中，底物曲率的效果将是 使用相同的聚合物和水凝胶底物进行了研究。理事假设是LEC迁移 EMT是由IOL植入后透镜胶囊平坦引起的较大曲率半径驱动的。 使用不同曲率半径的聚合物微图案评估曲率效应。玻璃 各种尺寸的微粒将嵌入水凝胶配方中，模仿镜头的变化 手术后的胶囊形状。在这两个目标中，都将使用相关的体外和离体模型。 LEC扩散， 将评估迁移和EMT的标记。 TGF-β和雷帕霉素将被用作阳性和阴性 EMT的诱导者。 RT-PCR将量化基因表达，蛋白质表达的变化将为 使用蛋白质印迹和免疫荧光评估。将评估的特定基因和蛋白质 包括SMAD信号蛋白，α-SMA，SLUG，蜗牛，纤连蛋白，E-钙粘蛋白和YAP。这个项目的目标 是为了确定底物机械性能（即粘弹性和曲率）如何有助于LEC EMT的行为和诱导。这将大大增强我们对LEC机械生物学的了解和 这些因素在EMT中的作用，提出了预防病理EMT的策略。结果将导致未来 研究材料设计以防止植入后EMT和纤维化的材料，特别是用于预防PCO。