scholarly journals Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations

Mutagenesis ◽  
2020 ◽  
Vol 35 (4) ◽  
pp. 319-330 ◽  
Author(s):  
Gillian E Conway ◽  
Ume-Kulsoom Shah ◽  
Samantha Llewellyn ◽  
Tereza Cervena ◽  
Stephen J Evans ◽  
...  

Abstract Following advancements in the field of genotoxicology, it has become widely accepted that 3D models are not only more physiologically relevant but also have the capacity to elucidate more complex biological processes that standard 2D monocultures are unable to. Whilst 3D liver models have been developed to evaluate the short-term genotoxicity of chemicals, the aim of this study was to develop a 3D model that could be used with the regulatory accepted in vitro micronucleus (MN) following low-dose, longer-term (5 days) exposure to engineered nanomaterials (ENMs). A comparison study was carried out between advanced models generated from two commonly used liver cell lines, namely HepaRG and HepG2, in spheroid format. While both spheroid systems displayed good liver functionality and viability over 14 days, the HepaRG spheroids lacked the capacity to actively proliferate and, therefore, were considered unsuitable for use with the MN assay. This study further demonstrated the efficacy of the in vitro 3D HepG2 model to be used for short-term (24 h) exposures to genotoxic chemicals, aflatoxin B1 (AFB1) and methyl-methanesulfonate (MMS). The 3D HepG2 liver spheroids were shown to be more sensitive to DNA damage induced by AFB1 and MMS when compared to the HepG2 2D monoculture. This 3D model was further developed to allow for longer-term (5 day) ENM exposure. Four days after seeding, HepG2 spheroids were exposed to Zinc Oxide ENM (0–2 µg/ml) for 5 days and assessed using both the cytokinesis-block MN (CBMN) version of the MN assay and the mononuclear MN assay. Following a 5-day exposure, differences in MN frequency were observed between the CBMN and mononuclear MN assay, demonstrating that DNA damage induced within the first few cell cycles is distributed across the mononucleated cell population. Together, this study demonstrates the necessity to adapt the MN assay accordingly, to allow for the accurate assessment of genotoxicity following longer-term, low-dose ENM exposure.

2001 ◽  
Vol 61 (2) ◽  
pp. 329-332 ◽  
Author(s):  
J. O. VALSA ◽  
I. FELZENSZWALB

Three tinctures samples from extracts of the popular medicinal plant Thuya occidentalis were tested in vitro through two short term tests for measuring the activity of genotoxic chemicals. Using the Salmonella/mammalian-microsome (Mutatest) assay and the SOS-chromotest (induction of beta-galactosidase in Escherichia coli), none of the extract was effective in inducing mutagenesis or beta-galactosidase synthesis (as an indicator of general and early sign of DNA damage), even with metabolization.


Mutagenesis ◽  
2010 ◽  
Vol 26 (1) ◽  
pp. 193-197 ◽  
Author(s):  
C. F. Bull ◽  
S. Beetstra-Hill ◽  
B. J. Benassi-Evans ◽  
J. W. Crott ◽  
M. Kimura ◽  
...  

2019 ◽  
Vol 26 (7) ◽  
pp. 512-522
Author(s):  
Xian Li ◽  
Long Xia ◽  
Xiaohui Ouyang ◽  
Qimuge Suyila ◽  
Liya Su ◽  
...  

<P>Background: Despite new agent development and short-term benefits in patients with Colorectal Cancer (CRC), metastatic CRC cure rates have not improved due to high rates of oxaliplatin resistance and toxicity. There is an urgent need for effective tools to prevent and treat CRC and reduce morbidity and mortality of CRC patients. Exploring the effects of bioactive peptides on the antitumor to CRC was of vital importance to the clinical application. </P><P> Objective: This study aimed to investigate the therapeutic impact of Anticancer Bioactive Peptides (ACBP) on anticancer effect of oxaliplatin (LOHP) in human colorectal cancer xenografts models in nude mice. </P><P> Methods: HCT-116 cells were cultured in vitro via CCK-8 assays and the absorbance was measured at 450 nm. Apoptosis and cell cycle were assessed by Flow Cytometry (FCM) in vitro. HCT-116 human colorectal cancer cells inoculated subcutaneously in nude mice of treatment with PBS (GG), ACBP, LOHP, ACBP+LOHP (A+L) in vivo. The quality of life was assessed by dietary amount of nude mice, the weight of nude mice, inhibition rates, tumor weight and tumor volume. Immunohistochemistry and RT-qPCR method was conducted to determine the levels of apoptosisregulating proteins/genes in transplanted tumors. </P><P> Results: ACBP induced substantial reductions in viable cell numbers and apoptosis of HCT116 cells in combined with LOHP in vitro. Compared with the control GG group, ACBP combined low dose oxaliplatin (U) group demonstrated significantly different tumor volume, the rate of apoptosis, the expression levels of Cyt-C, caspase-3,8,9 proteins and corresponding RNAs (P<0.05). The expression of pro-apoptotic proteins in the cytoplasm around the nucleus was significantly enhanced by ACBP. Short term intermittent use of ACBP alone indicted a certain inhibitory effect on tumor growth, and improve the quality of life of tumor bearing nude mice. </P><P> Conclusion: ACBP significantly increased the anti-cancer responses of low dose oxaliplatin (L-LOHP), thus, significantly improving the quality of life of tumor-bearing nude mice.</P>


2006 ◽  
pp. 135-142
Author(s):  
I. Lagroye ◽  
B. Wettring ◽  
E. G. Moros ◽  
W. L. Straube ◽  
W. F. Pickard ◽  
...  

2016 ◽  
Vol 258 ◽  
pp. S316
Author(s):  
D.J. Smart ◽  
F. Helbling ◽  
P. Blandine ◽  
M. Damian ◽  
V. Patrick

2020 ◽  
Vol 7 ◽  
pp. 1010-1019 ◽  
Author(s):  
David Thorne ◽  
James Whitwell ◽  
Julie Clements ◽  
Paul Walker ◽  
Damien Breheny ◽  
...  

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