Study of laser fluorescence spectroscopy in livers of rats with hypothermic ischemia.

Acta cirurgica brasileira Pub Date : 2023-12-01 eCollection Date: 2023-01-01 DOI:10.1590/acb386023
Allison Takeo Tsuge, Jaqueline de Jesus Pereira, José Dirceu Vollet-Filho, Márcia Saldanha Kubrusly, Flávio Henrique Ferreira Galvão, Orlando Nascimento Ribeiro, Camila Rodrigues Moreno, Renata Nishiyama Ikegami, Eleazar Chaib, Orlando de Castro E Silva
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Abstract

Purpose: After partial hepatectomy (PH), the remaining liver (RL) undergoes regenerative response proportional to the host. Limited literature exists on hepatic viability after tissue injury during hypothermic preservation. Spectroscopy measures cellular fluorescence and is explored for tissue characterization and parameter investigation. This study aimed to assess fluorescence analysis (spectroscopy) in evaluating liver viability and its relationship with hepatic tissue regeneration 24 hours after PH. Additionally, we analyzed liver regeneration in RL after 70% partial hepatectomy under hypothermic conditions with laser irradiation.

Methods: Fifty-six Wistar rats were divided into four groups: total non-perfused liver (control), total perfused liver, partial hepatectomy "in situ", and partial hepatectomy "ex situ". Tissue analysis was performed at 0 and 24 hours using spectroscopy with laser devices emitting at 532 (green) and 405 nm (violet).

Results: Spectroscopy identified tissue viability based on consistent results with Ki67 staining. The fluorescence spectra and Ki67 analysis displayed similar patterns, linking proliferative activity and absorption intensity.

Conclusions: Fluorescence spectroscopy proves to be promising for real-time analysis of cellular activity and viability. Metabolic activity was observed in groups of live animals and hypothermically preserved samples, indicating cellular function even under blood deprivation and hypothermic conditions.

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低温缺血大鼠肝脏激光荧光光谱研究。
目的:肝部分切除(PH)后,剩余肝脏(RL)发生与宿主成比例的再生反应。关于低温保存过程中组织损伤后肝脏活力的研究文献有限。光谱学测量细胞荧光,并探索组织表征和参数调查。本研究旨在评估荧光分析(光谱)在ph后24小时评估肝脏活力及其与肝组织再生的关系。此外,我们分析了低温条件下激光照射下70%肝部分切除术后RL的肝脏再生。方法:56只Wistar大鼠分为全肝未灌注组(对照组)、全肝灌注组(对照组)、原位肝部分切除术组(对照组)和非原位肝部分切除术组(对照组)。使用532(绿色)和405 nm(紫色)激光器件在0和24小时进行组织分析。结果:基于Ki67染色一致的结果,光谱学鉴定了组织活力。荧光光谱和Ki67分析显示出相似的模式,将增殖活性和吸收强度联系起来。结论:荧光光谱技术在实时分析细胞活性和活力方面具有广阔的应用前景。在活体动物和低温保存的样品中观察到代谢活性,表明即使在血液剥夺和低温条件下细胞功能。
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