Bioengineering of Transport Across Bruch's Membrance

跨布鲁赫膜运输的生物工程

• 批准号: 6743606
• 负责人: MARK JOHNSON
• 金额: $ 26.54万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6743606
• 关键词: basement membrane; bioengineering /biomedical engineering; biological fluid transport; choroid uvea; early diagnosis; electron microscopy; extracellular matrix; freeze etching; histopathology; human tissue; lipid transport; macular degeneration; membrane activity; membrane structure; morphometry; physical model; retinal pigment epithelium

DESCRIPTION (provided by applicant): Better understanding of the pathophysiology of the early stages of age-related maculopathy (ARM) may guide development of better treatments and/or preventions of age-related macular degeneration. Prominent clinical and histopathologic lesions of early ARM involve Bruch's membrane, a thin connective tissue between the choriocapillaris and the RPE. Unlike the case for late ARM, for which several risk factors have been identified, the only consistently identified risk factor for early ARM is advanced age. It is, therefore, important to understand how age-related changes in Bruch's membrane predispose some individuals for retinal dysfunction. A decreased transport capacity of Bruch's membrane, perhaps caused by lipid accumulation, has been postulated to be a critical early event in the pathogenesis of ARM. However, morphometric studies have not yet been successful in identifying those moieties responsible for limiting the transport capacity of Bruch's membrane. QFDE is a morphological technique that preserves extracellular matrix in exquisite detail and allows structures to be visualized that are not seen using conventional transmission electron microscopy (TEM) techniques. QFDE offers a technology that, in other connective tissues, has provided new insights into the transport characteristics of these tissues. Our goal is to combine the technique of quick-freeze/deep-etch (QFDE) morphology with bioengineering analytic methods to evaluate this hypothesis, using well-preserved eyes from normal human donors. Our specific aims are (i) to use the methods of quick-freeze/deep-etch (QFDE) morphology and lipid histochemistry in conjunction with conventional electron microscopy methods to characterize quantitatively the earliest ultrastructural changes in Bruch's membrane associated with lipid deposition in 20-70 year old donor eyes; (ii) to combine these results with analytical bioengineering tools to evaluate the effects of age-related changes on transport properties of Bruch's membrane; and (iii) to use an in vitro model of fluid and lipid transport through extracellular matrix to validate the results of the analytical studies and characterize the impact of lipid accumulation on transport.

描述（由申请人提供）：更好地了解与年龄相关的大斑（ARM）早期阶段的病理生理学，可以指导开发更好的治疗方法和/或预防与年龄相关的黄斑变性。早期手臂的突出的临床和组织病理病变涉及Bruch的膜，Bruch的膜是脉络膜毛细血管和RPE之间的薄结缔组织。与已确定多个危险因素的晚臂案件不同，早期手臂的唯一始终确定的危险因素是高龄。因此，重要的是要了解Bruch膜中与年龄相关的变化如何使某些个体患有视网膜功能障碍。 Bruch膜的运输能力降低，可能是由脂质积累引起的，被认为是ARM发病机理中的重要早期事件。但是，形态计量学研究尚未成功地识别那些负责限制Bruch膜运输能力的部分。 QFDE是一种形态学技术，可在精美的细节中保留细胞外基质，并允许使用常规传输电子显微镜（TEM）技术可视化结构。 QFDE提供了一项技术，该技术在其他结缔组织中为这些组织的运输特征提供了新的见解。我们的目标是使用普通人类捐助者保存完好的眼睛，将快速冻结/深蚀刻（QFDE）形态的技术结合到生物工程分析方法，以评估这一假设。 我们的具体目的是（i）使用快速冻结/深蚀刻（QFDE）形态和脂质组织化学与常规电子显微镜方法结合使用，以定量地表征BRUCH膜中最早的超微结构性变化，与20-70年的Donor Donor Eye gy Lipids相关的BRUCH's MEMBRANE与脂质沉积相关。 （ii）将这些结果与分析生物工程工具相结合，以评估与年龄相关的变化对BRUCH膜运输特性的影响； （iii）使用通过细胞外基质的液体和脂质转运的体外模型来验证分析研究的结果，并表征脂质积累对转运的影响。