Lab of Toxicology, Department of Health Sciences, The Graduate School of Dong-A University, 37, Nakdong-daero 550 Beon-gil, Saha-gu, Busan, 49315, Republic of Korea.
Division of Toxicological Research, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, Osong, Cheongju, 28159, Republic of Korea.
Regul Toxicol Pharmacol. 2022 Feb;128:105085. doi: 10.1016/j.yrtph.2021.105085. Epub 2021 Nov 17.
The development of a universal, label-free, and reliable in vitro toxicity testing method for nanoparticles is urgent because most nanoparticles can interfere with toxicity assays. In this regard, the colony-forming efficacy (CFE) assay has been suggested as a suitable in vitro toxicity assay for testing nanoparticles without such interference. Recently, the Organisation for Economic Co-operation and Development (OECD) developed a 60 × 15 mm Petri dish-based CFE assay for testing nanoparticles in MDCK-1 cells. However, further investigations are needed, including testing with other cell types, at a smaller scale for greater efficiency, and the application of the co-culture technique. In this study, we selected TiO, CuO, CeO, and SiO as test nanoparticles and successfully developed a 6-well plate-based CFE assay using HepG2 and A549 cells and a co-culture assay for combinations of HepG2 cells and THP-1 macrophages or A549 cells and THP-1 monocytes. The results suggest that the 6-wellplate-based CFE assay for HepG2 and A549 cells can be applied to nanoparticles, but the co-culture CFE assay has limitations in that it is not different from the single culture study, and it inhibits colony-formation by A549 cells in the presence of macrophages; this warrant further study.
开发一种通用的、无标记的、可靠的纳米粒子体外毒性测试方法是紧迫的，因为大多数纳米粒子会干扰毒性测定。在这方面，集落形成效率（CFE）测定被认为是一种合适的体外毒性测定方法，可用于测试无此类干扰的纳米粒子。最近，经济合作与发展组织（OECD）开发了一种基于 60×15mm 培养皿的 CFE 测定法，用于在 MDCK-1 细胞中测试纳米粒子。然而，需要进一步研究，包括使用其他细胞类型进行测试，以更小的规模提高效率，并应用共培养技术。在这项研究中，我们选择了 TiO2、CuO、CeO2 和 SiO2 作为测试纳米粒子，并成功地使用 HepG2 和 A549 细胞开发了一种基于 6 孔板的 CFE 测定法，以及 HepG2 细胞和 THP-1 巨噬细胞或 A549 细胞和 THP-1 单核细胞的共培养测定法。结果表明，基于 HepG2 和 A549 细胞的 6 孔板 CFE 测定法可用于纳米粒子，但共培养 CFE 测定法存在局限性，因为它与单一培养研究没有区别，并且在存在巨噬细胞的情况下抑制了 A549 细胞的集落形成；这需要进一步研究。
