Influence of serum on in situ proliferation and genotoxicity in A549 human lung cells exposed to nanomaterials

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• 作者：Sara Corradi^a ; ^{sacorrad@vub.ac.be} ; Laetitia Gonzalez^a ; Leen C.J. Thomassen^b ; Dagmar Bilani膷ová ; ^c ; Renie K. Birkedal^d ; Giulio Pojana^c ; Antonio Marcomini^c ; Keld A. Jensen^d ; Luc Leyns^a ; Micheline Kirsch-Volders^a
• 关键词：In situ ; Nanoparticles ; Toxicity ; Cytokinesis-blocked proliferation index ; Serum
• 刊名：Mutation Research/Genetic Toxicology and Environmental Mutagenesis
• 出版年：2012
• 期刊代码：31_13835718
• 类别：pha
• 出版时间：14 June, 2012
• 卷：745
• 期：1-2
• 页码：21-27
• 文件大小：795 K

摘要

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 渭g/ml and 50 渭g/ml, respectively and for Lys-SiO₂ NM at 3.5 渭g/ml in 0%serum. Increase in MN frequency was observed in cells treated in 10%serum with 50 渭g/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 渭g/ml. No toxicity was detected for TiO₂ and MWCNTs in both 10%and 0%serum, up to the dose of 250 渭g/ml. Restoration of CBPI comparable to untreated control was shown for cells cultured without serum and treated with 5 渭g/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.