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: 7653208
• 负责人: Robert J Levy
• 金额: $ 48.06万
• 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 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7653208
• 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新的方法，通过使用心脏瓣膜替换使用三叶叶瓣替换，使用三叶叶聚氨酯假体替换，这些假体是使用血液出生的内皮细胞（BOEC）进行内皮种子的。我们将研究由Abiomed（MA）制造的Abiocor聚氨酯阀（ABV）。 ABV已用于7,000多名人类受试者。假设BOEC种子ABV是有效抵抗聚氨酯心脏瓣膜衰竭的主要原因的有效性，其中包括血栓形成，钙化和氧化性降解小叶表面。目标1.结合并表征一种新型的两种成分光活化复合物，用于表面修饰胆固醇的聚氨酯心脏瓣膜小叶，以假设在高流量和高剪切的情况下假设增加BOEC粘附。 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组 - 该复合物的第二个组成部分称为CPB。这些研究的目的是创建一组候选PBPC-CPB公式，以在AIMS 2和3中进行调查。 AIM 2。对通过PBPC-CPB修饰的聚氨酯表面的体外研究，以促进BOEC粘附。这些研究将评估各种候选CPB的粘附机制，BOEC表型稳定性，内皮功能和激活。然后，我们将研究用铅CPB化合物修饰的ABV，并用二尖瓣流量模拟播种。综上所述，这些研究将用于了解候选PBPC-CPB的粘附机制和指导制剂的变化，这将在下面描述的AIM 3中导致最佳体内细胞粘附。 AIM 3。对BOEC播种的体内研究（使用绵羊），该研究将研究最佳的PBPC-CPB制剂。将研究两个体内模型系统：1）在2周的研究中，将评估BOEC粘附，表型稳定性和势力抗性的主动脉纽扣模型植入物。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，内皮激活以及血小板和纤维蛋白血栓沉积物。 2）用PBPC-CPB和BOEC种子和非转化对照修饰的ABV表面的二尖瓣替换，将进行150天，目的是评估BOEC粘附和表型稳定性，以及用于预防势头，钙化和氧化降解的治疗功效。公共卫生相关性：项目叙事心脏瓣膜疾病会影响数百万，只能通过心脏手术治疗。另外，由于血栓形成和主要材料失败，目前人造心脏瓣膜经常失败。该提案将通过研究用自体血外生长内皮细胞播种的三叶瓣聚氨酯阀来研究一种新的心脏瓣膜替代方法。