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Thermal Reduction of Graphene Oxide Mitigates Its In Vivo Genotoxicity Toward Xenopus laevis Tadpoles.
Evariste L
,
Lagier L
,
Gonzalez P
,
Mottier A
,
Mouchet F
,
Cadarsi S
,
Lonchambon P
,
Daffe G
,
Chimowa G
,
Sarrieu C
,
Ompraret E
,
Galibert AM
,
Ghimbeu CM
,
Pinelli E
,
Flahaut E
,
Gauthier L
.
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The worldwide increase of graphene family materials raises the question of the potential consequences resulting from their release in the environment and future consequences on ecosystem health, especially in the aquatic environment in which they are likely to accumulate. Thus, there is a need to evaluate the biological and ecological risk but also to find innovative solutions leading to the production of safer materials. This work focuses on the evaluation of functional group-safety relationships regarding to graphene oxide (GO) in vivo genotoxic potential toward X. laevis tadpoles. For this purpose, thermal treatments in H₂ atmosphere were applied to produce reduced graphene oxide (rGOs) with different surface group compositions. Analysis performed indicated that GO induced disturbances in erythrocyte cell cycle leading to accumulation of cells in G0/G1 phase. Significant genotoxicity due to oxidative stress was observed in larvae exposed to low GO concentration (0.1 mg.L-¹). Reduction of GO at 200 °C and 1000 °C produced a material that was no longer genotoxic at low concentrations. X-ray photoelectron spectroscopy (XPS) analysis indicated that epoxide groups may constitute a good candidate to explain the genotoxic potential of the most oxidized form of the material. Thermal reduction of GO may constitute an appropriate "safer-by-design" strategy for the development of a safer material for environment.
Figure 4. Cell-cycle distribution in G0/G1, S and G2/M phase analyzed from circulating erythrocytes of Xenopus laevis exposed to increasing concentrations of GO for 12 days. NC: negative control, N = 13, analysis of variance (ANOVA) p < 0.001 followed by Tukey test. Letters indicate significant differences between concentrations tested for each phase of the cell cycle.
Figure 5. Micronucleus induction measured in erythrocytes of Xenopus laevis larvae exposed for 12 days to GO or rGO (rGO200 or rGO1000). MNE: micronucleated erythrocytes; NC: negative control; PC: positive control; *: significant difference compared to the NC (McGill test).
Figure 1. Transmission electron microscopy micrographs of (A) Graphene oxide, (B) reduced graphene oxide at 200 °C, (C) reduced graphene oxide at 1000 °C.
Figure 2. X-ray photoelectron spectroscopy (XPS) survey spectra of GO, rGO200 and rGO1000 materials (A); C1s and O1s deconvoluted XPS spectra for GO, rGO200 and rGO1000 (B).
Figure 3. Monitoring of the stability of GO and rGO200 dispersion in the water column of exposure medium over 24 h (in absence of Xenopus larvae), expressed by the percentage of transmission detected after the light goes through the sample. Blank: medium without nanoparticles.
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