PDF(Pubmed)
Abstract：
BACKGROUND: Cardiopulmonary bypass is an essential component of cardiothoracic surgeries. However, significant complications such as systemic inflammatory response syndrome (SIRS) resulting from cardiopulmonary bypass (CPB) are a common occurrence due to contact between circulating blood and foreign surfaces that leads to platelet activation. It is suggested that different available CPB circuit coatings can potentially reduce platelet activation. However, there have been no published evidence-based reports confirming these claims. In addition, there is no well-established protocol for studying platelet activation biomarkers during CPB in vitro in a laboratory setting.
METHODS: CPB was simulated in the laboratory using bovine blood in two different types of coated CPB circuits: Trillium® Biosurface by Medtronic, and XcoatingTM Surface by Terumo. Fresh bovine blood samples were collected and circulated through the CPB circuit following the standard protocol used in the operation rooms. Blood samples were then collected at 5 min, 30 min, and 55 min during the circulation. Blood plasmas were separated and subjected to enzyme-linked immunosorbent assay to measure most established platelet activation markers P-selectin, Platelet Factor 4 (PF4), Glycoprotein IIb/IIIa (GPIIb/IIIa), and β-thromboglobulin (β-TG) at different time points.
RESULTS: The biomarker values at 30 min and 55 min were compared to the base values at 5 min for each type of CPB circuit. The results of the means from all measured biomarkers showed data measurements that indicated no significant variability within each coating. All collected data points fell within ±2 SD of the means, which was considered acceptable variations across technical replicates. Conclusion: In this study, we were able to establish an in vitro protocol in the laboratory setting that is precise and reliable with minimum intra-variability. This established protocol will allow for future studies in which different coated CPB circuits can be compared for their effectiveness in blocking platelet activation during the CPB.
METHODS: CPB was simulated in the laboratory using bovine blood in two different types of coated CPB circuits: Trillium® Biosurface by Medtronic, and XcoatingTM Surface by Terumo. Fresh bovine blood samples were collected and circulated through the CPB circuit following the standard protocol used in the operation rooms. Blood samples were then collected at 5 min, 30 min, and 55 min during the circulation. Blood plasmas were separated and subjected to enzyme-linked immunosorbent assay to measure most established platelet activation markers P-selectin, Platelet Factor 4 (PF4), Glycoprotein IIb/IIIa (GPIIb/IIIa), and β-thromboglobulin (β-TG) at different time points.
RESULTS: The biomarker values at 30 min and 55 min were compared to the base values at 5 min for each type of CPB circuit. The results of the means from all measured biomarkers showed data measurements that indicated no significant variability within each coating. All collected data points fell within ±2 SD of the means, which was considered acceptable variations across technical replicates. Conclusion: In this study, we were able to establish an in vitro protocol in the laboratory setting that is precise and reliable with minimum intra-variability. This established protocol will allow for future studies in which different coated CPB circuits can be compared for their effectiveness in blocking platelet activation during the CPB.
摘要：
背景：心肺转流术是心胸手术的重要组成部分。然而,体外循环(CPB)导致的全身炎症反应综合征(SIRS)等重大并发症是常见的,原因是循环血液与异物表面接触导致血小板活化.建议不同的可用CPB回路涂层可以潜在地减少血小板活化。然而,没有发表的基于证据的报告证实这些说法。此外,目前尚无在实验室环境中体外研究CPB期间血小板活化生物标志物的既定方案.
方法：在实验室中使用牛血在两种不同类型的涂层CPB回路中模拟CPB：Medtronic的Trillium®Biosurface，和Terumo的XcoatingTMSurface。按照手术室中使用的标准方案,收集新鲜的牛血液样品并循环通过CPB回路。然后在5分钟时收集血样，30分钟,和55分钟在循环过程中。分离血浆并进行酶联免疫吸附测定，以测量大多数已建立的血小板活化标志物P-选择素，血小板因子4(PF4),糖蛋白IIb/IIIa（GPIIb/IIIa），和β-血小板球蛋白（β-TG）在不同的时间点。
结果:对于每种类型的CPB回路,将30分钟和55分钟时的生物标志物值与5分钟时的基值进行比较。来自所有测量的生物标志物的平均值的结果显示,数据测量表明在每个涂层内没有显著的可变性。所有收集的数据点都落在平均值的±2SD内，这被认为是技术重复的可接受的变化。结论：在这项研究中，我们能够在实验室环境中建立精确可靠且内部变异性最小的体外方案.该建立的方案将允许未来的研究，其中可以比较不同的涂层CPB回路在CPB期间阻断血小板活化的有效性。
方法：在实验室中使用牛血在两种不同类型的涂层CPB回路中模拟CPB：Medtronic的Trillium®Biosurface，和Terumo的XcoatingTMSurface。按照手术室中使用的标准方案,收集新鲜的牛血液样品并循环通过CPB回路。然后在5分钟时收集血样，30分钟,和55分钟在循环过程中。分离血浆并进行酶联免疫吸附测定，以测量大多数已建立的血小板活化标志物P-选择素，血小板因子4(PF4),糖蛋白IIb/IIIa（GPIIb/IIIa），和β-血小板球蛋白（β-TG）在不同的时间点。
结果:对于每种类型的CPB回路,将30分钟和55分钟时的生物标志物值与5分钟时的基值进行比较。来自所有测量的生物标志物的平均值的结果显示,数据测量表明在每个涂层内没有显著的可变性。所有收集的数据点都落在平均值的±2SD内，这被认为是技术重复的可接受的变化。结论：在这项研究中，我们能够在实验室环境中建立精确可靠且内部变异性最小的体外方案.该建立的方案将允许未来的研究，其中可以比较不同的涂层CPB回路在CPB期间阻断血小板活化的有效性。
