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Shear Stress-Induced Total Blood Trauma in Multiple Species

机译：剪切应激诱导多种物种的总血小伤

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The common complications in heart failure patients with implanted ventricular assist devices (VADs) include hemolysis, thrombosis, and bleeding. These are linked to shear stress-induced trauma to erythrocytes, platelets, and von Willebrand factor (vWF). Novel device designs are being developed to reduce the blood trauma, which will need to undergo in vitro and in vivo preclinical testing in large animal models such as cattle, sheep, and pig. To fully understand the impact of device design and enable translation of preclinical results, it is important to identify any potential species-specific differences in the VAD-associated common complications. Therefore, the purpose of this study was to evaluate the effects of shear stress on cells and proteins in bovine, ovine, and porcine blood compared to human. Blood from different species was subjected to various shear rates (0-8000/s) using a rheometer. It was then analyzed for complete blood counts, hemolysis by the Harboe assay, platelet activation by flow cytometry, vWF structure by immunoblotting, and function by collagen binding activity ELISA (vWF : CBA). Overall, increasing shear rate caused increased total blood trauma in all tested species. This analysis revealed species-specific differences in shear-induced hemolysis, platelet activation, and vWF structure and function. Compared to human blood, porcine blood was the most resilient and showed less hemolysis, similar blood counts, but less platelet activation and less vWF damage in response to shear. Compared to human blood, sheared bovine blood showed less hemolysis, similar blood cell counts, greater platelet activation, and similar degradation of vWF structure, but less impact on its activity in response to shear. The shear-induced effect on ovine blood depended on whether the blood was collected via gravity at the abattoir or by venepuncture from live sheep. Overall, ovine abattoir blood was the least resilient in response to shear and bovine blood was the most similar to human blood. These results lay the foundations for developing blood trauma evaluation standards to enable the extrapolation of in vitro and in vivo animal data to predict safety and biocompatibility of blood-handling medical devices in humans. We advise using ovine venepuncture blood instead of ovine abattoir blood due to the greater overall damage in the latter. We propose using bovine blood for total blood damage in vitro device evaluation but multiple species could be used to create a full understanding of the complication risk profile of new devices. Further, this study highlights that choice of antibody clone for evaluating platelet activation in bovine blood can influence the interpretation of results from different studies.
机译：心力衰竭患者植入心室辅助装置（VADS）的常见并发症包括溶血，血栓形成和出血。这些与红细胞，血小板和von Willebrand因子（VWF）的剪切应激诱导的创伤有关。正在开发新颖的设备设计以减少血液创伤，这需要在大型动物模型（如牛，绵羊和猪）中进行体外和体内临床前测试。为了充分了解器件设计的影响并实现临床前效果的翻译，重要的是识别VAD相关常见并发症的任何潜在物种特异性差异。因此，本研究的目的是评估与人类相比牛，绵羊和猪血液中细胞和蛋白质对细胞和蛋白质的影响。使用流变仪对来自不同物种的血液进行各种剪切速率（0-8000 / s）。然后分析其完全血液计数，通过哈尔博测定溶血，通过流式细胞术，通过免疫印迹，VWF结构的血小板活化，并通过胶原蛋白结合活性ELISA（VWF：CBA）。总体而言，剪切速率的增加引起了所有测试物种中的总血小伤创伤。该分析揭示了剪切诱导溶血，血小板活化和VWF结构和功能的物种特异性差异。与人体血液相比，猪血是最有弹性的，并且表现出较少的溶血，类似的血液计数，但血小板活化较少，较少的VWF损伤响应剪切。与人体血液相比，剪切牛血液显示出较少的溶血，相似的血细胞计数，更高的血小板活化，以及VWF结构的类似降解，但响应于剪切而对其活性的影响较小。对绵血血液的剪切诱导的效果取决于血液是否通过Abattir的重力或由活羊的静脉穿刺收集。总体而言，绵羊AbattoR血液是抗剪切和牛血的最少的弹性与人体血液最相似。这些结果为开发血液创伤评估标准的基础奠定了开发血液创伤评价标准，以使体外和体内动物数据的外推，以预测人类血液处理医疗器械的安全性和生物相容性。我们建议使用卵巢静脉血血，而不是由于后者的总体损坏导致绵羊Abattoir血液。我们建议使用牛血来进行体外设备评估的总血液损伤，但多种物种可用于充分了解新设备的复杂性风险概况。此外，该研究突出显示用于评估牛血中的血小板活化的抗体克隆可以影响不同研究结果的解释。

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