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Fig. 3 | Journal of Nanobiotechnology

Fig. 3

From: Understanding the impact of more realistic low-dose, prolonged engineered nanomaterial exposure on genotoxicity using 3D models of the human liver

Fig. 3

Cytotoxicity and genotoxicity responses in HepG2 spheroids following both acute (24 h) and prolonged (120 h) exposure to 0.2–10.0 µg/mL of (A) TiO2, (B) ZnO, (C) Ag, (D) BaSO4 and (E) CeO2 ENMs. Cytotoxicity was assessed using the cytokinesis-block proliferation index (CBPI) for acute exposures, whilst trypan blue was assessed for the prolonged exposures, both of which are presented relative to the negative, untreated control. A known liver carcinogen, aflatoxin B1 (0.1 µM) was used as a positive control for genotoxicity. For acute exposures, 1000 binucleated cells were scored per replicate for each dose point using the cytokinesis-block version of the MN assay (2000 binucleate cells scored in total per dose). For prolonged exposures, 2000 mononucleated cells were scored per replicate for each dose point using the mononuclear MN assay (4000 mononucleate cells scored in total per dose). Mean data of two and three biological replicates (n = 2, n = 3) for genotoxicity and cytotoxicity respectively is presented ± SD. Significance indicated in relation to the negative control: * = p ≤ 0.05

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