In this work in situ proliferation of A549 human lung epithelial carcinoma cells exposed to nanomaterials (NMs) was investigated in the presence or absence of 10% serum. NMs were selected based on chemical composition, size, charge and shape (Lys-SiO2, TiO2, ZnO, and multi walled carbon nanotubes, MWCNTs). Cells were treated with NMs and 4 h later, cytochalasin-B was added. 36 h later, cell morphology was analyzed under a light microscope. Nuclearity was scored to determine the cytokinesis-block proliferation index (CBPI). CBPI, based on percentage of mono-, bi- and multi-nucleated cells, reflects cell toxicity and cell cycle delay. For some conditions depending on NM type (TiO2 and MWCNT) and serum concentration (0%) scoring of CBPI was impossible due to overload of agglomerated NMs. Moreover, where heavy agglomeration occurs, micronuclei (MN) detection and scoring under microscope was prevented. A statistically significant decrease of CBPI was found for ZnO NM suspended in medium in the absence or presence of 10% serum at 25 μg/ml and 50 μg/ml, respectively and for Lys-SiO2 NM at 3.5 μg/ml in 0% serum. Increase in MN frequency was observed in cells treated in 10% serum with 50 μg/ml ZnO. In 0% serum, the concentrations tested led to high toxicity. No genotoxic effects were induced by Lys-SiO2 both in the absence or presence of serum up to 5 μg/ml. No toxicity was detected for TiO2 and MWCNTs in both 10% and 0% serum, up to the dose of 250 μg/ml. Restoration of CBPI comparable to untreated control was shown for cells cultured without serum and treated with 5 μg/ml of Lys-SiO2 NM pre-incubated in 100% serum. This observation confirms the protective effect of serum on Lys-SiO2 NM cell toxicity. In conclusion in situ CBPI is proposed as a simple preliminary assay to assess both NMs induced cell toxicity and feasibility of MN scoring under microscope.
|Titolo:||Influence of serum on in situ proliferation and genotoxicity in A549 human lung cells exposed to nanomaterials|
|Data di pubblicazione:||2012|
|Appare nelle tipologie:||2.1 Articolo su rivista |
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|2011_CORRADI et al.Mut.Research.pdf||Post-print||Accesso chiuso-personale||Riservato|