Štampar Martina, Žegura Bojana
National Institute of Biology, Department of Genetic Toxicology and Cancer Biology, Večna pot 121, Ljubljana 1000, Slovenia.
National Institute of Biology, Department of Genetic Toxicology and Cancer Biology, Večna pot 121, Ljubljana 1000, Slovenia.
Mutat Res Genet Toxicol Environ Mutagen. 2024 Nov-Dec;900:503835. doi: 10.1016/j.mrgentox.2024.503835. Epub 2024 Nov 12.
The rapid development of new chemicals and consumer products has raised concerns about their potential genotoxic effects on human health, including DNA damage leading to serious diseases. For such new chemicals and pharmaceutical products, international regulations require genotoxicity data, initially obtained through in vitro tests, followed by in vivo experiments, if needed. Traditionally, laboratory animals have been used for this purpose, however, they are costly, ethically problematic, and often unreliable due to species differences. Therefore, innovative more accurate in vitro testing approaches are rapidly being developed to replace, refine and reduce (3R) the use of animals for experimental purposes and to improve the relevance for humans in toxicology studies. One of such innovative approaches are in vitro three-dimensional (3D) cell models, which are already being highlighted as superior alternatives to the two-dimensional (2D) cell cultures that are traditionally used as in vitro models for the safety testing of chemicals and pharmaceuticals. 3D cell models provide physiologically relevant information and more predictive data for in vivo conditions. In the review article, we provide a comprehensive overview of 3D hepatic cell models, including HepG2, HepG2/C3A, HepaRG, human primary hepatocytes, and iPSC-derived hepatocytes, and their application in the field of genotoxicology. Through a detailed literature analysis, we identified 31 studies conducted between 2007 and April 2024 that used a variety of standard methods, such as the comet assay, the micronucleus assay, and the γH2AX assay, as well as new methodological approaches, including toxicogenomics, to assess the cytotoxic and genotoxic activity of chemicals, nanoparticles and natural toxins. Based on our search, we can conclude that the use of in vitro 3D cell models for genotoxicity testing has been increasing over the years and that 3D cell models have an even greater potential for future implementation and further refinement in genetic toxicology and risk assessment.
新化学品和消费品的快速发展引发了人们对其对人类健康潜在遗传毒性影响的担忧，包括导致严重疾病的DNA损伤。对于此类新化学品和药品，国际法规要求提供遗传毒性数据，最初通过体外试验获得，如有必要，随后进行体内实验。传统上，实验动物一直用于此目的，然而，它们成本高昂、存在伦理问题，并且由于物种差异往往不可靠。因此，正在迅速开发创新的、更准确的体外测试方法，以替代、优化和减少（3R原则）用于实验目的的动物使用，并提高毒理学研究中对人类的相关性。此类创新方法之一是体外三维（3D）细胞模型，它已被视为传统上用作化学品和药品安全性测试体外模型的二维（2D）细胞培养的优越替代品。3D细胞模型为体内条件提供生理相关信息和更具预测性的数据。在这篇综述文章中，我们全面概述了3D肝细胞模型，包括HepG2、HepG2/C3A、HepaRG、人原代肝细胞和诱导多能干细胞衍生的肝细胞，以及它们在遗传毒理学领域的应用。通过详细的文献分析，我们确定了2007年至2024年4月期间进行的31项研究，这些研究使用了多种标准方法，如彗星试验、微核试验和γH2AX试验，以及包括毒理基因组学在内的新方法，以评估化学品、纳米颗粒和天然毒素的细胞毒性和遗传毒性活性。基于我们的搜索，我们可以得出结论，多年来，体外3D细胞模型在遗传毒性测试中的使用一直在增加，并且3D细胞模型在遗传毒理学和风险评估的未来实施和进一步优化方面具有更大的潜力。
