Pharmaceutical Analysis (XB20), Groningen Research Institute of Pharmacy, University of Groningen, 9713 AV Groningen, The Netherlands.
Pharmaceutical Technology and Biopharmacy, Groningen Research Institute of Pharmacy, University of Groningen, 9713 AV Groningen, The Netherlands.
Anal Chem. 2024 Oct 8;96(40):15871-15879. doi: 10.1021/acs.analchem.4c00355. Epub 2024 Sep 29.
We developed an organ-on-a-chip (OOC) based on precision-cut liver slices to assess liver function in real-time, both in health and disease, in a controlled and noninvasive manner. We achieved this by integrating fiber-optic-based oxygen sensors before and after the microchamber in which a liver slice was incubated under flow, to measure oxygen concentrations in the medium in real time. We first demonstrated that the basal oxygen consumption rate (OCR) of liver slices is a reliable indicator of liver slice viability. By monitoring basal OCR (2.9-5.7 pmol O/min/μg protein) in incubation medium, we found that it correlated well to cellular adenosine triphosphate (ATP) content (3.0-7.9 pmol/μg protein) ( = 0.82, < 0.0001). Second, we induced a diseased state in liver slices by targeting the mitochondria, as they play a critical role in liver function and disease. We exposed the liver slices to succinate in abundance (40 mM) for short periods (1 h) to rapidly boost mitochondrial OCR. Two successive treatments of succinate increased the OCR of liver slices by 1.5 pmol/min/μg each time. However, between treatments, the liver slice OCR did not return to its basal OCR, instead decreasing drastically by 60-70%, suggesting succinate toxicity. We confirmed this with ATP analysis (1.0 pmol/μg protein) and hematoxylin and eosin staining, which showed tissue necrosis and apoptosis. Our system could be an advantageous model for future studies assessing liver (patho)physiology in response to potentially toxic drugs or lifestyle-related liver diseases.
我们开发了一种基于微流控芯片的器官芯片（OOC）系统，用于实时评估健康和疾病状态下的肝功能，其方式为精准、微创且可控制。我们通过在孵育微流控芯片前后整合光纤氧传感器来实现这一点，该传感器可以实时测量孵育微流控芯片中肝切片的氧浓度。首先，我们证明了肝切片的基础耗氧量（OCR）是肝切片活力的可靠指标。通过监测孵育介质中的基础 OCR（2.9-5.7 pmol O/min/μg 蛋白），我们发现其与细胞三磷酸腺苷（ATP）含量（3.0-7.9 pmol/μg 蛋白）高度相关（ = 0.82， < 0.0001）。其次，我们通过靶向线粒体来诱导肝切片产生疾病状态，因为线粒体在肝功能和疾病中起着关键作用。我们将肝切片暴露在丰富的琥珀酸盐（40 mM）中较短时间（1 小时），以快速提高线粒体的耗氧量。两次连续的琥珀酸盐处理使肝切片的 OCR 每次增加 1.5 pmol/min/μg。然而，在两次处理之间，肝切片的 OCR 并没有恢复到基础 OCR，反而急剧下降 60-70%，表明琥珀酸盐毒性。我们通过 ATP 分析（1.0 pmol/μg 蛋白）和苏木精-伊红染色证实了这一点，结果显示组织坏死和细胞凋亡。我们的系统可能成为未来研究的有利模型，用于评估可能有毒的药物或与生活方式相关的肝脏疾病对肝脏（病理）生理学的影响。
