Watanabe Nobuo, Sakota Daisuke, Ohuchi Katsuhiro, Takatani Setsuo
Department of Artificial Organs, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
Artif Organs. 2007 May;31(5):352-8. doi: 10.1111/j.1525-1594.2007.00392.x.
In this study, mechanical trauma to red blood cells was evaluated by conventional hemolysis test and a newly developed cyclically reversing shear flow generator. The fresh porcine blood obtained from a local slaughterhouse was subjected to the conventional hemolysis test using a commercial centrifugal blood pump for the duration of 8 h. The measurements consisted of (i) plasma-free hemoglobin based on the standard optical measurement and (ii) the deformability of red blood cells (RBCs) using a cyclically reversing shear flow generator and microscope image acquisition system. The deformability of RBCs was expressed by the L/W value where L and W were the longer and shorter axes of the elongated RBCs' images. Although the plasma-free hemoglobin level increased with the pumping duration, the L/W remained unchanged for the duration of 8 h of pumping to indicate no alteration in the deformability. It was speculated that (i) although RBCs might have been circulated for so many times through the test pump, after each exposure to mechanical stress, RBCs might have recovered, and net effect due to shear stress-exposure time might have been small; and (ii) RBCs' deformability might be maintained near normal until sudden burst or membrane rupture, or the hemoglobin might have continuously leaked through the pores of the thinned membrane created by the mechanical stress. The deformability testing under a fluctuating shear flow could be a new method to quantify subhemolytic mechanical damage that has been accumulated in the RBCs' membrane and that may not be assessed by the conventional hemolysis test.
在本研究中,通过传统溶血试验和新开发的循环反向剪切流发生器评估红细胞的机械损伤。从当地屠宰场获取的新鲜猪血,使用商用离心血泵进行8小时的传统溶血试验。测量内容包括:(i)基于标准光学测量的无血浆血红蛋白;(ii)使用循环反向剪切流发生器和显微镜图像采集系统测量红细胞(RBC)的变形性。RBC的变形性用L/W值表示,其中L和W分别是拉长的RBC图像的长轴和短轴。尽管无血浆血红蛋白水平随泵送时间增加,但在泵送8小时期间L/W保持不变,表明变形性没有改变。据推测:(i)尽管RBC可能多次通过测试泵循环,但每次受到机械应力后,RBC可能已经恢复,并且由于剪切应力暴露时间产生的净效应可能很小;(ii)RBC的变形性可能在突然破裂或膜破裂之前保持接近正常,或者血红蛋白可能持续通过机械应力造成的变薄膜的孔隙泄漏。在波动剪切流下进行变形性测试可能是一种量化RBC膜中累积的亚溶血机械损伤的新方法,而传统溶血试验可能无法评估这种损伤。