Bioengineering Design of Artificial Blood

人工血液的生物工程设计

• 批准号: 7682841
• 负责人: Marcos Intaglietta
• 金额: $ 71.31万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-05 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7682841
• 关键词: Acids; Affinity; Amines; Animal Model; Animals; Apoptosis; Binding; Biochemical; Biological Assay; Biomedical Engineering; Blood; Blood Circulation; Blood Substitutes; Blood Transfusion; Blood Vessels; Blood Volume; Blood capillaries; Caliber; Carbon Monoxide; Cardiac; Cardiac Output; Cell Death; Characteristics; Chloride Ion; Chlorides; Clinical; Clinical Trials; Collaborations; Colloids; Conscious; Coronary Occlusions; Data; Development; Diffusion; Digestion; Dose; Drug Formulations; Effectiveness; Endothelium; Endotoxins; Engineering; Equation; Erythrocytes; Europe; Freezing; Gases; Generic Drugs; Glycocalyx; Goals; Hamsters; Heart; Heart Rate; Heme; Hemodilution; Hemoglobin; Hemorrhage; High Pressure Liquid Chromatography; Homeostasis; Hour; Human; Hydration status; Infarction; Ischemia; Laboratories; Length; Ligand Binding; Ligands; Lysine; Maintenance; Maleimides; Measurement; Measures; Medicine; Membrane Proteins; Metabolic; Methods; Microcirculation; Military Personnel; Modeling; Molecular Conformation; Molecular Models; Molecular Weight; Monitor; Myocardial Infarction; Myocardial Ischemia; Names; Necrosis; Nitric Oxide; Nitrite Reductase; Osmolalities; Osmotic Pressure; Oxygen; PEG-hemoglobin; Phase; Phase II Clinical Trials; Phase III Clinical Trials; Physiological; Plasma; Polyethylene Glycols; Polymers; Preparation; Procedures; Production; Property; Proteins; Radial; Rattus; Reaction; Renal function; Reperfusion Therapy; Research; Research Personnel; Research Project Grants; Research Proposals; Resuscitation; Role; Safety; Site; Solutions; Staging; Structure; Sulfhydryl Compounds; Surface; Sweden; System; TNFRSF5 gene; Temperature; Test Result; Testing; Thermodynamics; Time; Toxic effect; Transfusion; Transport Process; Trauma; Vasodilator Agents; Viscosity; Weight; Work; alkyl group; awake; base; capillary; chemical synthesis; clinical application; crosslink; density; design; hemodynamics; improved; in vivo; light scattering; mathematical model; methyl 4-mercaptobutyrimidate; molecular dynamics; molecular modeling; molecular size; myocardial infarct sizing; novel; oxygen transport; pressure; repaired; research study; resistance factors; simulation; tissue oxygenation; triphenyltetrazolium; vasoconstriction

In the first period, work in this BRP demonstrated that MP4 (PEG-modified hemoglobin) overcomes the most significant hurdle to the development of modified hemoglobin-based blood substitutes, namely vasoconstriction. MP4 promotes tissue oxygenation through a combination of O2 transport and maintenance of functional capillary density in spite of its counterintuitive properties, including increased O2 affinity, viscosity and oncotic pressure. MP4 is superior to blood in its ability to resuscitate animals from severe, uncontrolled hemorrhage, and it has been shown to be safe in human clinical trials. In this present application we will test the hypothesis that MP4 is an effective carrier of the heme ligands O2, carbon monoxide (CO)and nitric oxide (NO), and will carry out the related physiological studies in order to understand its effectiveness as a blood substitute and identify new clinical applications for its use. We will test the hypothesis that PEG-Hb formulations are vasodilators which interact with the circulation in ways not related to NO scavenging by Hb. We propose that MP4's properties are in part due to an increased nitrite reductase activity and NO transport. We will develop a procedure for using MP4 to deliver CO and exploit that CO-MP4 is exceptionally stable, even at elevated temperatures, making it valuable in field use for trauma. Project 1 has a GMP facility that provides MP4 of consistent quality and properties. It will develop and produce new PEG-Hb compounds with goals to optimize concentration without increasing colloid osmotic pressure and augment O2 delivery capacity so as to increase the applicability of MP4 to a wider range of clinical uses. Properties will be screened via biochemical analysis, mathematical modeling, and systemic experiments in rats including the effect on myocardial infarction. Project 2 will examine PEG-Hbs' effects on the glycocalyx integrity, how PEG-Hbs' presence influences reactive O2 species (ROS), and will investigate the combined effect of PEG-Hb and enhanced plasma viscosity. MP4 will be used as a delivery vehicle for CO to provide cellular protection during ischemia & hemorrhage. Microcirculation studies will use the awake hamster window chamber model, with direct measurements of O2 and NO levels, flow and diameter in blood vessels and functional capillary density. This research combines physiological analysis of transfusion, fundamentals of engineering transport processes and mechanotransduction with the expertise of two laboratories with more than 12 years of collaboration. Effective blood substitutes will significantly increase the safety and efficacy of blood transfusions in civilian and military settings, streamline and simplify transfusion medicine. Applications for a new type of blood substitute will be developed, one that provides a fundamentally different treatment of ischemia, based on enhancement of microvascular flow, cardiac and renal functions, repair of the endothelium and delivery of heme ligands (O2, NO and CO).

在第一阶段，该 BRP 的工作证明 MP4（PEG 修饰的血红蛋白）克服了大多数 开发基于血红蛋白的改良血液替代品的重大障碍，即 血管收缩。 MP4 通过 O2 运输和维持的结合促进组织氧合 功能性毛细血管密度，尽管其具有违反直觉的特性，包括增加 O2 亲和力， 粘度和胶体渗透压。 MP4 比血液更能帮助动物从严重的、 不受控制的出血，并且在人体临床试验中已被证明是安全的。在这个当下 应用中我们将测试 MP4 是血红素配体 O2、碳的有效载体的假设 一氧化碳（CO）和一氧化氮（NO），并将开展相关的生理学研究，以便 了解其作为血液替代品的有效性并确定其新的临床应用。我们将 检验 PEG-Hb 制剂是血管舒张剂的假设，其与循环的相互作用方式并非如此 与 Hb 清除 NO 相关。我们认为 MP4 的特性部分归因于亚硝酸盐的增加 还原酶活性和 NO 转运。我们将开发一个使用 MP4 传送 CO 并利用的程序 即使在高温下，CO-MP4 也非常稳定，这使得它在现场使用中很有价值 创伤。项目 1 拥有 GMP 设施，可提供质量和性能稳定的 MP4。将会发展 并生产新的 PEG-Hb 化合物，目标是在不增加胶体的情况下优化浓度 渗透压并增强 O2 输送能力，从而增加 MP4 的适用范围 临床用途范围。将通过生化分析、数学建模和筛选特性 大鼠全身实验，包括对心肌梗塞的影响。项目 2 将检查 PEG-Hbs' 对糖萼完整性的影响，PEG-Hbs 的存在如何影响活性氧 (ROS)，以及 研究 PEG-Hb 和增强血浆粘度的综合效应。 MP4将用作交付 CO 的载体，在缺血和出血期间提供细胞保护。微循环研究将使用 清醒仓鼠窗室模型，可直接测量 O2 和 NO 水平、流量和 血管直径和功能性毛细血管密度。这项研究结合了生理分析 输血、工程运输过程的基础知识和机械转导的专业知识 两个实验室合作超过 12 年。有效的血液替代品将显着 提高民用和军事环境中输血的安全性和有效性，简化和简化 输血医学。将开发一种新型血液替代品的应用，该替代品可以提供 缺血的根本不同治疗方法是基于微血管血流、心脏和血管的增强 肾功能、内皮修复和血红素配体（O2、NO 和 CO）的输送。