Endothelial Progenitor Cell Seeded Self-Expanding Vascular Stent

内皮祖细胞接种的自膨式血管支架

• 批准号: 8175947
• 负责人: Hardean Eric Achneck
• 金额: $ 19.63万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-05 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8175947
• 关键词: Adhesions; Affect; Affinity Chromatography; Angioplasty; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Autologous; Binding; Blood Vessels; Cell Wall; Cell physiology; Cell surface; Cells; Characteristics; Devices; Endothelial Cells; Environment; Extracellular Matrix Proteins; Family suidae; Glycocalyx; Goals; Human body; Image; Implant; Incubated; Knowledge; Label; Laboratories; Lasers; Left; Lesion; Life; Liquid substance; Measures; Mediating; Metals; Methodology; Methods; Microscopy; Nitric Oxide; Nitrites; Obstruction; Patients; Performance; Peripheral; Peripheral Vascular Diseases; Phenotype; Phosphopeptides; Phosphoproteins; Physiological; Population; Procedures; Production; Property; Research; Risk; Sampling; Scanning; Seeds; Side; Solutions; Solvents; Stem cells; Stents; Surface; System; Technology; Testing; Thrombomodulin; Thrombosis; Time; Translating; Vascular Diseases; Venous; Western Blotting; external jugular vein; implantation; improved; in vivo; innovation; new technology; nitinol; novel; peripheral blood; prevent; restenosis; shear stress; titanium dioxide

DESCRIPTION (provided by applicant): Objectives: It is the goal of this study to develop a technology to line self-expanding Nitinol stents with autologous peripheral blood-derived endothelial progenitor cells (EPCs) in order to reduce the common and potentially devastating complications of in-stent restenosis and thrombosis. Nitinol stents are used to treat narrowing of blood vessels in patients with peripheral vascular disease, an illness that will affect over 20 million patients in the U.S. by 2020. Aims and Methods: It is our Aim I in this study to optimize a novel technology to seed Nitinol stents with EPCs derived from peripheral blood and to evaluate EPC function on the stent surface ex vivo. We plan on manufacturing an innovative Nitinol delivery system that allows seeding the stents within minutes of implantation by laser drilling micropores in the outside stent sheath so that EPCs can be seeded on the inside strut surface by using the principle of solvent drag when forcing EPC-containing solution through a stent side- port. We will also optimize stent imaging and re-compressing methods. In Aim I.B. we will assess cell retention on Nitinol stents under physiological shear stress in an ex vivo flow circuit and test whether adhesion is mediated by cell wall phosphoproteins. In Aim I.C. we evaluate nitric oxide production by quantifying nitrite in media samples with an Ionics/Sievers Nitric Oxide Analyzer and thrombomodulin expression by EPCs on the stent surface with Western blot analysis under different flow conditions. In Aim 2, we will test EPC-coated Nitinol stents in a porcine animal model. Aim 2.A. will allow us to test our novel methods of seeding Nitinol stents with fluorescently-labeled EPCs and implanting them into 3 pilot animals in a short-term survival study. Following, in Aim 2.B. we will implant one EPC-coated and one uncoated (control) stent into the right and left external jugular vein of 7 pigs. The differences in thrombosis and restenosis between treated and control stents will be evaluated by histomorphometric semiquantitative analyses, as well as scanning- and fluorescent microscopy. Impact: The knowledge gained in this proposal will deepen our understanding of the function and adhesion mechanism of EPCs on Nitinol under shear stress. If successful, we will have provided proof-of-concept for a novel technology, which will significantly improve the performance of self-expanding stents and may benefit millions of patients. PUBLIC HEALTH RELEVANCE: The goal of this research is to develop a novel technology in order to avoid the dangerous and life-threatening clogging of Nitinol stents. In order to achieve this goal we aim to coat stents with EPCs immediately before implanting them in into the human body. If successful, this will render stents free from obstructions - thus benefitting millions of patients.

描述（由申请人提供）： 目的：本研究的目标是开发一种技术，将自膨式镍钛诺支架与自体外周血来源的内皮祖细胞（EPC）排列在一起，以减少支架内再狭窄和血栓形成的常见且可能具有破坏性的并发症。镍钛合金支架用于治疗周围血管疾病患者的血管狭窄，这种疾病到 2020 年将影响美国超过 2000 万患者。 目的和方法：本研究的目的 I 是优化一种新技术，将外周血中的 EPC 植入镍钛合金支架，并离体评估支架表面的 EPC 功能。我们计划制造一种创新的镍钛诺输送系统，通过在支架外鞘上激光钻出微孔，可以在植入后几分钟内对支架进行播种，以便在迫使含有 EPC 的溶液通过支架侧端口时，利用溶剂拖曳原理将 EPC 播种到支柱内表面。我们还将优化支架成像和重新压缩方法。在《目标》中，I.B.我们将评估离体流动回路中生理剪切应力下镍钛诺支架上的细胞保留，并测试粘附是否是由细胞壁磷蛋白介导的。在 Aim I.C.我们通过使用 Ionics/Sievers 一氧化氮分析仪定量培养基样品中的亚硝酸盐来评估一氧化氮的产生，并通过蛋白质印迹分析在不同的流动条件下评估支架表面 EPC 的血栓调节蛋白表达。在目标 2 中，我们将在猪动物模型中测试 EPC 涂层镍钛合金支架。目标 2.A.将使我们能够测试我们的新方法，即在镍钛诺支架上植入荧光标记的 EPC，并将其植入 3 只试点动物体内进行短期生存研究。接下来，在目标 2.B 中。我们将在 7 头猪的左右颈外静脉中植入一个 EPC 涂层支架和一个未涂层支架（对照）。将通过组织形态半定量分析以及扫描和荧光显微镜来评估治疗支架和对照支架之间血栓形成和再狭窄的差异。影响：本提案中获得的知识将加深我们对剪切应力下镍钛合金上 EPC 的功能和粘附机制的理解。如果成功，我们将为一项新技术提供概念验证，该技术将显着提高自膨胀支架的性能，并可能使数百万患者受益。 公共健康相关性：这项研究的目标是开发一种新技术，以避免镍钛合金支架发生危险且危及生命的堵塞。为了实现这一目标，我们的目标是在将支架植入人体之前立即为其涂上 EPC。如果成功，这将使支架畅通无阻，从而使数百万患者受益。