Optical fluorescence spectroscopy to detect hepatic necrosis after normothermic ischemia: animal model

R. A. Romano, J. D. Vollet-Filho, S. Pratavieira, J. L. Fernández, C. Kurachi, V. Bagnato, O. Castro-e-Silva, A. Sankarankutty
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Abstract

Liver transplantation is a well-established treatment for liver failure. However, the success of the transplantation procedure depends on liver graft conditions. The tissue function evaluation during the several transplantation stages is relevant, in particular during the organ harvesting, when a decision is made concerning the viability of the graft. Optical fluorescence spectroscopy is a good option because it is a noninvasive and fast technique. A partial normothermic hepatic ischemia was performed in rat livers, with a vascular occlusion of both median and left lateral lobes, allowing circulation only for the right lateral lobe and the caudate lobe. Fluorescence spectra under excitation at 532 nm (doubled frequency Nd:YAG laser) were collected using a portable spectrometer (USB2000, Ocean Optics, USA). The fluorescence emission was collected before vascular occlusion, after ischemia, and 24 hours after reperfusion. A morphometric histology analysis was performed as the gold standard evaluation ─ liver samples were analyzed, and the percentage of necrotic tissue was obtained. The results showed that changes in the fluorescence emission after ischemia can be correlated with the amount of necrosis evaluated by a morphometric analysis, the Pearson correlation coefficient of the generated model was 0.90 and the root mean square error was around 20%. In this context, the laser-induced fluorescence spectroscopy technique after normothermic ischemia showed to be a fast and efficient method to differentiate ischemic injury from viable tissues.
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荧光光谱法检测常温缺血后肝坏死的动物模型
肝移植是一种公认的治疗肝衰竭的方法。然而,移植手术的成功取决于肝移植的条件。在几个移植阶段的组织功能评估是相关的,特别是在器官采集期间,当决定移植物的生存能力时。光学荧光光谱是一个很好的选择,因为它是一种无创和快速的技术。在大鼠肝脏中进行部分常温肝缺血,中间和左外侧叶血管闭塞,只允许右外侧叶和尾状叶循环。使用便携式光谱仪(USB2000, Ocean Optics, USA)采集532 nm(双频Nd:YAG激光)激发下的荧光光谱。在血管闭塞前、缺血后和再灌注后24小时采集荧光发射。进行形态计量学组织学分析作为金标准评估──分析肝脏样本,并获得坏死组织的百分比。结果表明,荧光发射在缺血后的变化可与形态学分析评估的坏死程度相关,生成的模型的Pearson相关系数为0.90,均方根误差在20%左右。在此背景下,常温缺血后激光诱导荧光光谱技术是一种快速有效的区分缺血损伤和活组织的方法。
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