Schmidt Konrad, Berg Johanna, Roehrs Viola, Kurreck Jens, Al-Zeer Munir A
Department of Applied Biochemistry, Institute of Biotechnology, 4/3-2, Technische Universität Berlin, Gustav-Meyer-Allee 25, 13355 Berlin, Germany.
Toxicol Rep. 2020 Nov 21;7:1578-1587. doi: 10.1016/j.toxrep.2020.11.003. eCollection 2020.
In recent years 3D-bioprinting technology has been developed as an alternative to animal testing. It possesses a great potential for testing as it aims to mimic human organs and physiology. In the present study, an alginate-gelatin-Matrigel based hydrogel was used to prepare 3D-bioprinted HepaRG cultures using a pneumatic extrusion printer. These 3D models were tested for viability and metabolic functions. Using 3D-bioprinted HepaRG cultures, we tested the toxicity of aflatoxin B1 (10 or 20 μM) and compared the results with 2D HepaRG cultures. There was a dose-dependent toxicity effect on cell viability, reduction of metabolic activity and albumin production. We found that 3D-bioprinted HepaRG cultures are more resistant to aflatoxin B1 treatment than 2D cultures. Although the metabolic activities were reduced upon treatment with aflatoxin B1, the 3D models were still viable and survived longer, up to 3 weeks, than the 2D culture, as visualized by fluorescence microscopy. Furthermore, albumin production recovered slightly in 3D models after one and two weeks of treatment. Taken together, we consider using 3D-bioprinting technology to generate 3D tissue models as an alternative way to study toxicity and this could also provide a suitable alternative for chronic hepatotoxicity studies .
近年来，3D生物打印技术已被开发出来作为动物实验的替代方法。它在测试方面具有巨大潜力，因为其旨在模拟人体器官和生理机能。在本研究中，一种基于藻酸盐 - 明胶 - 基质胶的水凝胶被用于使用气动挤压打印机制备3D生物打印的HepaRG细胞培养物。对这些3D模型进行了活力和代谢功能测试。利用3D生物打印的HepaRG细胞培养物，我们测试了黄曲霉毒素B1（10或20μM）的毒性，并将结果与二维HepaRG细胞培养物进行了比较。对细胞活力、代谢活性降低和白蛋白产生存在剂量依赖性毒性作用。我们发现，3D生物打印的HepaRG细胞培养物比二维培养物对黄曲霉毒素B1处理更具抗性。尽管在用黄曲霉毒素B1处理后代谢活性降低，但3D模型仍然存活，并且比二维培养物存活时间更长，长达3周，这通过荧光显微镜观察可见。此外，在处理1周和2周后，3D模型中的白蛋白产生略有恢复。综上所述，我们认为使用3D生物打印技术生成3D组织模型作为研究毒性的替代方法，这也可为慢性肝毒性研究提供合适的替代方案。
