Research Supplement to Promote Diversity Supplement for the ECMO without Anticoagulation

项目摘要/摘要 一氧化氮(NO)是一种由内皮细胞一氧化氮持续产生的气态自由基 合酶(ENOS)是血管壁上的一种酶，是 保持体内平衡，防止血液凝块形成。NO是一种活性极强的分子， 半衰期只有2ms，并能在血液中形成各种代谢物。血液的人造表面 接触设备可以激活血小板并诱导血栓形成。这是一种不受欢迎的 体外生命支持(ECLS)电路的副作用，通常由全身预防 抗凝，但这会增加出血并发症的风险。因此，有一个 努力创造更具生物相容性的人造表面，以消除对系统性疾病的需求 抗凝剂。 没有发射的人造曲面为这个问题提供了有吸引力的解决方案。然而，没有有效负载的 这样的表面本质上是有限的。 在本研究附录中，我们想要探索一种替代方法，在这种情况下，可用的 没有有效载荷以NO代谢物的形式在血液中循环(例如，亚硝酸盐，小的和 大分子S-亚硝硫醇)作为储集层，与 生物医学装置的化学修饰的人造表面。我们希望实现这一目标 通过以下目标： 目的1：建立血液中循环NO的检测方法。 将开发选择性分析方法来量化潜在的循环NO供体(亚硝酸盐， 血液中的小分子和大分子S亚硝硫醇)。 目的2：调节血液中循环NO库的浓度。这一目标是为了 评估输送的NO气体对体内单个NO代谢物数量的影响 鲜血。不会通过体外膜氧合器将气体输送到预适应的血液中 设置。这将允许精确控制NO向血液的输送(NO和 O2，扫气和血液的流速)。 目的3：对EC表面进行修饰，使其能够从循环的NO储存库中释放NO。 我们将在聚氯乙烯管材上涂上掺杂/共价改性的二价金属离子聚合物涂层 (Cu2+)，并评估其诱导NO从循环中的NO库释放的能力和防止 Chandler环模型体外血栓形成的实验研究。

PROJECT SUMMARY/ABSTRACT Nitric Oxide (NO) is a gaseous radical continuously produced by the endothelial nitric oxide synthase (eNOS) enzyme within the walls of the blood vessels and is an essential molecule to maintain homeostasis and prevent blood clot formation. NO is an extremely reactive molecule with only 2 ms half-life and can form various metabolites in the blood. The artificial surfaces of blood contacting devices can activate platelets and induce thrombus formation. This is a not desired side effect of Extracorporeal Life Support (ECLS) circuits and is typically prevented by systemic anticoagulation, which however increases the risk of hemorrhagic complications. Hence, there is an effort to create more biocompatible artificial surfaces that obviate the need for systemic anticoagulation. NO emitting artificial surfaces offer attractive solution to this problem. However, the NO payload of such surfaces is inherently limited. In this research supplement we would like to explore an alternative approach, where the available NO payload is circulating within the blood in the form of NO metabolites (e.g., nitrite, small and large molecular weight S-nitroso thiols) as reservoirs and is released upon contact with the chemically modified artificial surface of the biomedical device. We would like to achieve this goal through the following aims: Aim 1: Develop analytical methods to measure circulating NO donors in the blood. Selective analytical methods will be developed to quantify potential circulating NO donors (nitrite, small and large molecular weight S-nitroso thiols) in the blood. Aim 2: Modulate concentration of circulating NO reservoirs in the blood. This aim is to assess the effect of the delivered NO gas on the quantity of individual NO metabolites within the blood. NO gas will be delivered to pre-conditioned blood via an in vitro membrane oxygenator setup. This will allow for precise control of the delivery of NO to the blood (partial pressure of NO and O2, flow rate of sweep gas and blood). Aim 3: Modify EC surfaces that can induce NO release from circulating NO reservoirs. We will coat PVC tubing with a polymer coating doped/covalently modified with divalent metal ions (Cu2+) and assess its ability to induce NO release from the circulating NO reservoirs and to prevent blood clot formation in vitro in Chandler loop model.