PDF(Pubmed)
Abstract：
The current Organisation for Economic Co-Operation and Development test guideline number 487 (OECD TG No. 487) provides instruction on how to conduct the in vitro micronucleus assay. This assay is one of the gold standard approaches for measuring the mutagenicity of test items; however, it is directed at testing low molecular weight molecules and may not be appropriate for particulate materials (e.g. engineered nanoparticles [ENPs]). This study aimed to adapt the in vitro micronucleus assay for ENP testing and underpins the development of an OECD guidance document. A harmonized, nano-specific protocol was generated and evaluated by two independent laboratories. Cell lines utilized were human lymphoblastoid (TK6) cells, human liver hepatocytes (HepG2) cells, Chinese hamster lung fibroblast (V79) cells, whole blood, and buffy coat cells from healthy human volunteers. These cells were exposed to reference ENPs from the Joint Research Council (JRC): SiO2 (RLS-0102), Au5nm and Au30nm (RLS-03, RLS-010), CeO2 (NM212), and BaSO4 (NM220). Tungsten carbide-cobalt (WC/Co) was used as a trial particulate positive control. The chemical controls were positive in all cell cultures, but WC/Co was only positive in TK6 and buffy coat cells. In TK6 cells, mutagenicity was observed for SiO2- and both Au types. In HepG2 cells, Au5nm and SiO2 showed sub-two-fold increases in micronuclei. In V79 cells, whole blood, and buffy coat cells, no genotoxicity was detected with the test materials. The data confirmed that ENPs could be tested with the harmonized protocol, additionally, concordant data were observed across the two laboratories with V79 cells. WC/Co may be a suitable particulate positive control in the in vitro micronucleus assay when using TK6 and buffy coat cells. Detailed recommendations are therefore provided to adapt OECD TG No. 487 for testing ENP.
摘要：
当前的经济合作与发展组织第487号测试指南(OECDTGNo.487)提供了如何进行体外微核试验的指导。该测定法是测量测试项目致突变性的金标准方法之一；然而，它旨在测试低分子量分子,并且可能不适用于颗粒材料(例如工程纳米颗粒[ENPs])。这项研究旨在适应ENP测试的体外微核试验，并支持OECD指导文件的开发。一个和谐的，纳米特异性方案由两个独立的实验室生成和评估.使用的细胞系是人淋巴母细胞（TK6）细胞，人肝细胞（HepG2）细胞，中国仓鼠肺成纤维细胞（V79）细胞，全血,和健康人类志愿者的血沉棕黄层细胞。这些细胞暴露于来自联合研究委员会（JRC）的参考ENPs：SiO2（RLS-0102），Au5nm和Au30nm(RLS-03,RLS-010),CeO2(NM212),和BaSO4(NM220)。碳化钨-钴(WC/Co)用作试验颗粒阳性对照。所有细胞培养物中的化学对照均为阳性,但WC/Co仅在TK6和血沉棕黄层细胞中呈阳性。在TK6细胞中，观察到SiO2-和两种Au类型的致突变性。在HepG2细胞中，Au5nm和SiO2显示微核的亚两倍增加。在V79细胞中，全血,还有血沉棕黄层细胞,试验材料未检测到遗传毒性。数据证实ENPs可以用协调协议进行测试，此外，在使用V79细胞的两个实验室中观察到了一致的数据.当使用TK6和血沉棕黄层细胞时，WC/Co可能是体外微核试验中合适的微粒阳性对照。因此，提供了详细的建议，以适应经合组织TGNo.487用于测试ENP。
