Genetic Toxicity Assessment: Employing the Best Science for Human Safety Evaluation Part II: Performances of the In Vitro Micronucleus Test Compared to the Mouse Lymphoma Assay and the In Vitro Chromosome Aberration Assay

Lorge, Elisabeth; Lambert, Carine; Gervais, Véronique; Bécourt-Lhote, Nathalie; Delongeas, Jean-Luc; Claude, Nancy

doi:10.1093/toxsci/kfl193

Cited by 25 publications

(8 citation statements)

References 9 publications

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“…This activity could be confirmed in the present study with metabolic activation at a rather high cytotoxic concentration (relative total growth rate of 20% relative to the control). Generally, this assay system is known for a rather high rate of false positive responses, especially at cytotoxic concentrations (Caldwell, 1993;Lorge et al, 2007;Parry et al, 2010;Reeve et al, 2012). In other in vitro and in vivo genotoxicity assays, eugenol shows mixed results, although most of them are negative (EFSA, 2009).…”

Section: In Vitro Mammalian Cell Genotoxicitymentioning

confidence: 99%

Toxicological assessment of kretek cigarettes part 4: Mechanistic investigations, smoke chemistry and in vitro toxicity

Roemer

Dempsey

Lawless-Pyne

et al. 2014

Regulatory Toxicology and Pharmacology

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The smoke chemistry and in vitro toxicity of mainstream smoke (MS) was investigated in American-blended cigarettes with or without the addition of 2.5%, 5% or 10% eugenol to the tobacco and in Indonesian-blended cigarettes with and without the addition of cloves, cloves extracted with hot ethanol, and extracted cloves replenished with eugenol or clove oil. The addition of eugenol reduced the concentration of nearly all toxicants measured in MS as well as the in vitro cytotoxicity of the gas/vapor phase. Reductions were also seen in bacterial mutagenicity of the total particulate matter (TPM) assessed by the Ames Assay. The addition of extracted cloves led to increases and decreases of toxicant concentrations in MS. Replenishment with eugenol or clove oil decreased the toxicant concentrations; with most smoke constituent concentrations reduced below the concentration found in tobacco-only cigarettes. Cytotoxicity of the TPM was not affected by the clove preparations. However, GVP cytotoxicity was reduced (untreated cloves showing the highest reductions). Mutagenicity of TPM was decreased by the clove preparations. Mechanisms for the reductions, (up to 40%), are most likely due to dilution effects by eugenol, changed burning characteristics of the tobacco, and free radical scavenging by eugenol.

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Section: In Vitro Mammalian Cell Genotoxicitymentioning

confidence: 99%

Toxicological assessment of kretek cigarettes part 4: Mechanistic investigations, smoke chemistry and in vitro toxicity

Roemer

Dempsey

Lawless-Pyne

et al. 2014

Regulatory Toxicology and Pharmacology

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“…The in vitro micronucleus test is widely used as a screening assay to assess cytogenetic damage in mammalian cells [1][2][3][4][5]. Its popularity is largely due to the fact that it can be executed with less technical expertise relative to chromosome aberration analyses, and because it is capable of detecting aneugens as well as clastogens.…”

Section: Introductionmentioning

confidence: 99%

Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

Bryce

Avlasevich

Bemis

et al. 2008

Mutation Research/Genetic Toxicology and Environmental Mutagene

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An international, multi-lab trial was conducted to evaluate a flow cytometry-based method for scoring micronuclei in mouse lymphoma L5178Y cells [Avlasevich et al., Environ. Molec. Mutagen. 47 (2006) [56][57][58][59][60][61][62][63][64][65][66]. A reference laboratory investigated the potential of six chemicals to induce micronuclei-the genotoxicants mitomycin C, etoposide, and vinblastine, and the non-genotoxicants sucrose, staurosporine, or dexamethasone. The latter two non-genotoxicants were selected as extreme challenges to the assay because of their potent apoptogenic activity. Three collaborating laboratories were supplied with prototype In Vitro MicroFlow ™ kits, and each was assigned one genotoxicant and one non-genotoxicant. Cells were treated continuously for 24 hrs over a range of concentrations up to 5 mg/ml, or overtly cytotoxic concentrations. Micronuclei were scored via standard microscopy and flow cytometry. In addition to enumerating micronucleus frequencies, a cytotoxicity measurement that is simultaneously acquired with the flow cytometric micronucleus scoring procedure was evaluated (Flow-NBR). With this method, latex particles served as counting beads, and facilitated relative survival measurements that exclude the presence of dead/dying cells. For comparison purposes, additional cytotoxicity endpoints were measured, including several that are based on cell number, and others that reflect compromised membrane integrity, including dye permeability and/or phospholipid distribution. Key findings for this set of compounds include the following: (1) significant discrepancies in top concentration selection were found when cytotoxicity measurements were based on different methods, with the Flow-NBR approach tending to be the most sensitive, (2) both microscopy-and flow cytometry-based scoring methods detected concentrationdependent micronucleus formation for the three genotoxic agents studied, with good agreement between the reference laboratory and the collaborating laboratories, and (3) whereas flow cytometric analyses showed no significant increases for the non-genotoxicants when top concentration selection was based on Flow-NBR, significantly elevated micronucleus frequencies were observed for concentrations that were chosen based on less-sensitive cytotoxicity assays. Collectively, these results indicate that rapid assessment of genotoxicity can be accomplished with a relatively simple Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access

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“…The results of chromosome aberration test indicate that there was no significant difference between the different dosage groups and the negative control group(p>0.05), which meant that the samples would not cause chromosome aberration. The in vitro micronucleus test started to be used in genotoxicity in the 1970s [15].It can detect chromosomal damage in a further stage [16]. It is also a reliable method for evaluat¬ing the effect of genetic toxicity on chromosomes [17].…”

Section: Discussionmentioning

confidence: 99%

Study on tissue compatibility and genotoxicity of a radiation sterilized disposable burette transfusion apparatus

Chen

Liu

2011

Proceedings 2011 International Conference on Human Health and Biomedical Engineering

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The aim of the study was to evaluate the effect of radiation sterilization on biocompatibility of a disposable burette transfusion apparatus, in order to assess the safety of radiation sterilized medical devices. The initial bacteria burden of the burette transfusion apparatus was examed according to the ISO11737 standard, we combined cytotoxicity test, sensitization test and dermal irritation test to evaluate the tissue compatibility of the radiation sterilized samples, and the genotoxicity was investigated with chromosomal aberration test and micronucleus test. The study determined the verified sterilizing dosage at the level of 9.4kGy, results show the sample products had no cytotoxicity, no sensitization,no intracutaneous irritation, no genotoxicity in chromosome aberration test, and will not increase the micronucleus rate of PCE (p<0.05). Disposable burette transfusion apparatus showed good tissue compatibility and had no genotoxicity after radiation sterilization. Radiation sterilization is a biologically safe method for sterilization of medical devices.

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Genetic Toxicity Assessment: Employing the Best Science for Human Safety Evaluation Part II: Performances of the In Vitro Micronucleus Test Compared to the Mouse Lymphoma Assay and the In Vitro Chromosome Aberration Assay

Cited by 25 publications

References 9 publications

Toxicological assessment of kretek cigarettes part 4: Mechanistic investigations, smoke chemistry and in vitro toxicity

Toxicological assessment of kretek cigarettes part 4: Mechanistic investigations, smoke chemistry and in vitro toxicity

Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay

Study on tissue compatibility and genotoxicity of a radiation sterilized disposable burette transfusion apparatus

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