Blood Outgrowth Endothelial Cell Seeding of Heart Valve Leaflets

心脏瓣膜小叶的血液生长内皮细胞接种

• 批准号: 7894729
• 负责人: Robert J Levy
• 金额: $ 47.77万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7894729
• 关键词: Adhesions; Affect; Angioflex; Artificial Heart; Autologous; Biological Assay; Biological Models; Blood; Cardiac Surgery procedures; Cell Adhesion; Cell Separation; Cells; Cholesterol; Complex; Cotyledon plant; Deposition; Drug Formulations; Endothelial Cells; Exposure to; Failure; Fibrin; Flow Cytometry; Goals; Heart Valve Diseases; Heart Valve Prosthesis; Heart Valves; Hour; Implant; In Vitro; Investigation; Lead; Learning; Link; Methodology; Methods; Mitral Valve; Modeling; Modification; Phenotype; Platelet Activation; Polyurethanes; Procedures; Prosthesis; Protocols documentation; Publishing; Reporting; Research; Research Design; Resistance; Resources; Sheep; Side; Sulfhydryl Compounds; Surface; System; Thrombosis; Thrombus; Treatment Efficacy; Work; aortic valve replacement; base; calcification; cell type; heart valve replacement; human subject; in vivo; in vivo Model; mitral valve replacement; novel; novel strategies; polyallylamine; prevent; public health relevance; simulation

DESCRIPTION (provided by applicant): The proposed research will study a novel approach for treating heart valve disease via the use of heart valve replacements with trileaflet polyurethane prostheses that are endothelial seeded using blood outgrowth endothelial cells (BOEC). We will investigate the AbioCor polyurethane valve (ABV), manufactured by Abiomed (Danvers, MA). The ABV has been used in more than 7,000 human subjects. BOEC seeded ABV are hypothesized to be efficacious for resisting the principal causes of polyurethane heart valve failure, which include thrombosis, calcification and oxidative degradation of leaflet surfaces. Aim 1.Synthesize and characterize a novel two component photoactivated complex for surface modifying polyurethane heart valve leaflets with cholesterol to hypothetically increase BOEC adhesion under high flow and high shear. Our synthetic methods will use a two step approach using: 1) A photo- activatable polyallylamine-benzophone compound, referred to as PBPC, to activate the surface of polyurethane heart valve leaflets for the subsequent attachment to the photo-linked PBPC of, 2) a cholesterol-derivatized polyallylamine, biochemically modified with thiol-reactive groups and ionogenic groups--this 2nd component of the complex is referred to as CPB. The goal of these studies will be to create a candidate set of PBPC-CPB formulations for investigation in Aims 2&3. Aim 2. In vitro investigations of polyurethane surfaces modified with PBPC-CPB to promote BOEC adhesion. These studies will assess adhesion mechanisms, BOEC phenotype stability, endothelial function and activation with various candidate CPB. We will then study ABV modified with lead CPB compounds and BOEC seeded with mitral valve flow simulations. Taken together, these studies will be used to both understand adhesion mechanisms and guide formulation changes in candidate PBPC-CPBs that will result in optimal in vivo cell adhesion in Aim 3 described below. Aim 3. In vivo studies (using sheep) of BOEC seeding that will investigate optimal PBPC-CPB formulations. Two in vivo model systems will be studied: 1) An aortic button model implant that will assess BOEC adhesion, phenotype stability and thrombo-resistance in 2 week studies. Explants will also be studied for extent of cell coverage, endothelial phenotype stability, the presence of nonendothelial cell types, endothelial activation, and platelet and fibrin thrombus depositon. 2) Mitral valve replacements in sheep with ABV surface modified with PBPC-CPB and BOEC seeded and nonmodified controls will be carried out for 150 days with the goal of assessing both BOEC adhesion and phenotype stability, and therapeutic efficacy for preventing thrombus, calcification and oxidative degradation. PUBLIC HEALTH RELEVANCE: Project Narrative Heart valve disease affects millions and is only treatable by cardiac surgery. In addition current artificial heart valves fail frequently due to thrombosis and primary material failure. This proposal will investigate a novel approach for a heart valve replacement by studying a trileaflet polyurethane valve that is seeded with autologous blood outgrowth endothelial cells.

描述（由申请人提供）：拟议的研究将研究一种治疗心脏瓣膜疾病的新方法，通过使用血液外生内皮细胞（BOEC）内皮种子的三叶聚氨酯假体进行心脏瓣膜置换术。我们将研究Abiomed （Danvers， MA）生产的AbioCor聚氨酯阀门（ABV）。ABV已经在超过7000人身上使用。BOEC种子ABV被认为可以有效抵抗聚氨酯心脏瓣膜衰竭的主要原因，包括血栓形成、钙化和小叶表面的氧化降解。目的1。合成并表征了一种新型的双组分光活化复合物，用于高流量、高剪切下对含胆固醇的聚氨酯心脏瓣膜小叶进行表面改性，从而增加BOEC的粘附。我们的合成方法将采用两步方法：1)光活化聚烯丙胺-苯甲酮化合物，简称PBPC，激活聚氨酯心脏瓣膜小叶的表面，使其随后附着在光连接的PBPC上；2)胆固醇衍生化聚烯丙胺，用硫醇反应基团和离子生成基团进行生化修饰——该复合物的第二个组分称为CPB。这些研究的目标是为目标2和目标3的研究创建一套候选PBPC-CPB配方。目标2。PBPC-CPB改性聚氨酯表面促进BOEC粘附的体外研究。这些研究将评估粘附机制、BOEC表型稳定性、内皮功能和各种候选CPB的激活。然后，我们将研究含铅CPB化合物修饰的ABV和含二尖瓣流动模拟的BOEC。综上所述，这些研究将用于理解粘附机制，并指导候选PBPC-CPBs的配方变化，从而在目标3中实现最佳的体内细胞粘附。目标3。BOEC种子的体内研究（使用绵羊）将研究最佳PBPC-CPB配方。两种体内模型系统将被研究：1)主动脉钮扣模型植入物将在2周的研究中评估BOEC粘附、表型稳定性和血栓抵抗性。还将研究外植体的细胞覆盖范围、内皮表型稳定性、非内皮细胞类型的存在、内皮活化以及血小板和纤维蛋白血栓沉积。2)用PBPC-CPB修饰ABV表面的绵羊进行二尖瓣置换术，用BOEC种子和未修饰的对照组进行150天的二尖瓣置换术，目的是评估BOEC的粘附性和表型稳定性，以及预防血栓、钙化和氧化降解的治疗效果。心脏瓣膜疾病影响数百万人，只能通过心脏手术治疗。此外，目前的人工心脏瓣膜经常因血栓形成和原发性材料失效而失效。本提案将研究一种心脏瓣膜置换的新方法，通过研究一种用自体血液外生内皮细胞播种的三叶聚氨酯瓣膜。