Reconstructed human intestinal comet assay, a possible alternative in vitro model for genotoxicity assessment.

IF 2.5 4区 医学 Q3 GENETICS & HEREDITY
Mutagenesis Pub Date : 2023-06-20 DOI:10.1093/mutage/gead011
Christopher Owen Hughes, Hui Kheng Lim, Joseph Choon Wee Tan, David Ian Leavesley, Benjamin Paul Chapman Smith
{"title":"Reconstructed human intestinal comet assay, a possible alternative in vitro model for genotoxicity assessment.","authors":"Christopher Owen Hughes,&nbsp;Hui Kheng Lim,&nbsp;Joseph Choon Wee Tan,&nbsp;David Ian Leavesley,&nbsp;Benjamin Paul Chapman Smith","doi":"10.1093/mutage/gead011","DOIUrl":null,"url":null,"abstract":"<p><p>The aim of the present study was to evaluate the compatibility of reconstructed 3D human small intestinal microtissues to perform the in vitro comet assay. The comet assay is a common follow-up genotoxicity test to confirm or supplement other genotoxicity data. Technically, it can be performed utilizing a range of in vitro and in vivo assay systems. Here, we have developed a new reconstructed human intestinal comet (RICom) assay protocol for the assessment of orally ingested materials. The human intestine is a major site of food digestion and adsorption, first-pass metabolism as well as an early site of toxicant first contact and thus is a key site for evaluation. Reconstructed intestinal tissues were dosed with eight test chemicals: ethyl methanesulfonate (EMS), ethyl nitrosourea (ENU), phenformin hydrochloride (Phen HCl), benzo[a]pyrene (BaP), 1,2-dimethylhydrazine hydrochloride (DMH), potassium bromate (KBr), glycidamide (GA), and etoposide (Etop) over a span of 48 h. The RICom assay correctly identified the genotoxicity of EMS, ENU, KBr, and GA. Phen HCl, a known non-genotoxin, did not induce DNA damage in the 3D reconstructed intestinal tissues whilst showing high cytotoxicity as assessed by the assay. The 3D reconstructed intestinal tissues possess sufficient metabolic competency for the successful detection of genotoxicity elicited by BaP, without the use of an exogenous metabolic system. In contrast, DMH, a chemical that requires liver metabolism to exert genotoxicity, did not induce detectable DNA damage in the 3D reconstructed intestinal tissue system. The genotoxicity of Etop, which is dependent on cellular proliferation, was also undetectable. These results suggest the RICom assay protocol is a promising tool for further investigation and safety assessment of novel ingested materials. We recommend that further work will broaden the scope of the 3D reconstructed intestinal tissue comet assay and facilitate broader analyses of genotoxic compounds having more varied modes of actions.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":"38 3","pages":"139-150"},"PeriodicalIF":2.5000,"publicationDate":"2023-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10281391/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mutagenesis","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1093/mutage/gead011","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0

Abstract

The aim of the present study was to evaluate the compatibility of reconstructed 3D human small intestinal microtissues to perform the in vitro comet assay. The comet assay is a common follow-up genotoxicity test to confirm or supplement other genotoxicity data. Technically, it can be performed utilizing a range of in vitro and in vivo assay systems. Here, we have developed a new reconstructed human intestinal comet (RICom) assay protocol for the assessment of orally ingested materials. The human intestine is a major site of food digestion and adsorption, first-pass metabolism as well as an early site of toxicant first contact and thus is a key site for evaluation. Reconstructed intestinal tissues were dosed with eight test chemicals: ethyl methanesulfonate (EMS), ethyl nitrosourea (ENU), phenformin hydrochloride (Phen HCl), benzo[a]pyrene (BaP), 1,2-dimethylhydrazine hydrochloride (DMH), potassium bromate (KBr), glycidamide (GA), and etoposide (Etop) over a span of 48 h. The RICom assay correctly identified the genotoxicity of EMS, ENU, KBr, and GA. Phen HCl, a known non-genotoxin, did not induce DNA damage in the 3D reconstructed intestinal tissues whilst showing high cytotoxicity as assessed by the assay. The 3D reconstructed intestinal tissues possess sufficient metabolic competency for the successful detection of genotoxicity elicited by BaP, without the use of an exogenous metabolic system. In contrast, DMH, a chemical that requires liver metabolism to exert genotoxicity, did not induce detectable DNA damage in the 3D reconstructed intestinal tissue system. The genotoxicity of Etop, which is dependent on cellular proliferation, was also undetectable. These results suggest the RICom assay protocol is a promising tool for further investigation and safety assessment of novel ingested materials. We recommend that further work will broaden the scope of the 3D reconstructed intestinal tissue comet assay and facilitate broader analyses of genotoxic compounds having more varied modes of actions.

Abstract Image

Abstract Image

Abstract Image

重建人类肠道彗星试验,一种可能替代体外遗传毒性评估模型。
本研究的目的是评估重建的三维人小肠微组织的相容性,以进行体外彗星试验。彗星试验是一种常见的后续遗传毒性试验,用于确认或补充其他遗传毒性数据。从技术上讲,它可以利用一系列体外和体内分析系统进行。在这里,我们开发了一种新的重建人类肠道彗星(RICom)测定方案,用于评估口服摄入的物质。人体肠道是食物消化、吸附、首过代谢的主要部位,也是毒物最早接触的部位,是评价的关键部位。重建的肠道组织在48小时内分别加入8种化学物质:甲磺酸乙酯(EMS)、亚硝基脲乙酯(ENU)、盐酸苯双胍(Phen HCl)、苯并[a]芘(BaP)、盐酸1,2-二甲基肼(DMH)、溴酸钾(KBr)、甘油酯(GA)和乙油苷(Etop)。RICom实验正确鉴定了EMS、ENU、KBr和GA的遗传毒性。Phen HCl是一种已知的非基因毒素,在3D重建的肠组织中不会引起DNA损伤,但通过该检测显示出高细胞毒性。3D重建的肠道组织具有足够的代谢能力,可以成功检测BaP引起的遗传毒性,而无需使用外源性代谢系统。相比之下,DMH(一种需要肝脏代谢才能发挥遗传毒性的化学物质)在3D重建的肠组织系统中没有引起可检测到的DNA损伤。Etop的遗传毒性依赖于细胞增殖,也未检测到。这些结果表明,RICom检测方案是一种有前途的工具,用于进一步调查和安全评估新的摄入物质。我们建议进一步的工作将扩大3D重建肠组织彗星测定的范围,并促进具有更多不同作用模式的基因毒性化合物的更广泛分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Mutagenesis
Mutagenesis 生物-毒理学
CiteScore
5.90
自引率
3.70%
发文量
22
审稿时长
6-12 weeks
期刊介绍: Mutagenesis is an international multi-disciplinary journal designed to bring together research aimed at the identification, characterization and elucidation of the mechanisms of action of physical, chemical and biological agents capable of producing genetic change in living organisms and the study of the consequences of such changes.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信