Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro.
Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro.
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DOI：
10.1186/s12951-021-00769-9
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发表时间：
2021-01-19
影响因子：
10.2
通讯作者：
Doak SH
中科院分区：
文献类型：
--
作者：
Burgum MJ;Clift MJD;Evans SJ;Hondow N;Tarat A;Jenkins GJ;Doak SH
工程纳米材料（ENM）的毒理学评价对于职业健康和安全至关重要，特别是在涉及少层石墨烯（FLG）等批量制造的ENM时。此外，有必要在测试ENM时开发先进的体外模型，以提供侵入性动物实验的生理相关替代方案。本研究的目的是确定非功能化（中性），胺和羧基功能化FLG对人转化I型（TT 1）肺泡上皮细胞单培养物，以及TT 1和分化的THP-1单核细胞（d.THP-1（巨噬细胞））的共培养物的遗传毒性。在单一培养物中，TT 1和d.THP-1巨噬细胞在暴露24小时后对每种ENM显示出统计学显著（p < 0.05）的细胞毒性反应。通过体外胞质分裂阻断微核（CBMN）试验测量的单培养遗传毒性显示，8 µg/ml的胺-和羧基-FLG具有显著（p < 0.05）微核诱导作用。透射电子显微镜（TEM）显示ENM被膜结合囊泡内的TT 1细胞内化。在共培养物中，ENM在没有细胞毒性作用的情况下诱导遗传毒性。预暴露于1.5 mM N-乙酰半胱氨酸（NAC）的共培养物显示出微核诱导的基线水平，表明观察到的遗传毒性是由氧化应激引起的。因此，当在单一培养物中检查时，FLG遗传毒性导致初级-间接DNA损伤;而共培养细胞揭示了DNA损伤的次级机制。
Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs to provide a physiologically relevant alternative to invasive animal experimentation. The aim of this study was to determine the genotoxicity of non-functionalised (neutral), amine- and carboxyl-functionalised FLG upon both human-transformed type-I (TT1) alveolar epithelial cell monocultures, as well as co-cultures of TT1 and differentiated THP-1 monocytes (d.THP-1 (macrophages)). In monocultures, TT1 and d.THP-1 macrophages showed a statistically significant (p < 0.05) cytotoxic response with each ENM following 24-h exposures. Monoculture genotoxicity measured by the in vitro cytokinesis blocked micronucleus (CBMN) assay revealed significant (p < 0.05) micronuclei induction at 8 µg/ml for amine- and carboxyl-FLG. Transmission electron microscopy (TEM) revealed ENMs were internalised by TT1 cells within membrane-bound vesicles. In the co-cultures, ENMs induced genotoxicity in the absence of cytotoxic effects. Co-cultures pre-exposed to 1.5 mM N-acetylcysteine (NAC), showed baseline levels of micronuclei induction, indicating that the genotoxicity observed was driven by oxidative stress. Therefore, FLG genotoxicity when examined in monocultures, results in primary-indirect DNA damage; whereas co-cultured cells reveal secondary mechanisms of DNA damage.
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影响因子：
17.1
作者：
Ruenraroengsak P;Chen S;Hu S;Melbourne J;Sweeney S;Thorley AJ;Skepper JN;Shaffer MS;Tetley TD;Porter AE
通讯作者：
Porter AE
DOI：
10.1039/c5en00238a
发表时间：
2016-06-01
期刊：
Environmental science. Nano
影响因子：
--
作者：
Luanpitpong S;Wang L;Davidson DC;Riedel H;Rojanasakul Y
通讯作者：
Rojanasakul Y
DOI：
10.1016/j.mrgentox.2011.09.013
发表时间：
2012-06-14
影响因子：
1.9
作者：
Doak, S. H.;Manshian, B.;Jenkins, G. J. S.;Singh, N.
通讯作者：
Singh, N.
影响因子：
10
作者：
Wills JW;Hondow N;Thomas AD;Chapman KE;Fish D;Maffeis TG;Penny MW;Brown RA;Jenkins GJ;Brown AP;White PA;Doak SH
通讯作者：
Doak SH
影响因子：
5.6
作者：
Gurunathan, Sangiliyandi;Kang, Min-Hee;Kim, Jin-Hoi
通讯作者：
Kim, Jin-Hoi