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Suitability of cell-based label-free detection for cytotoxicity screening of carbon nanotubes.

Meindl C, Absenger M, Roblegg E, Fröhlich E - Biomed Res Int (2013)

Bottom Line: For validation of the label-free systems different concentrations of ethanol and of amine (AMI) polystyrene NPs were used.All systems identified thin (<8 nm) CNTs as more cytotoxic than thick (>20 nm) CNTs, but detection by xCELLigence system was less sensitive to CNT-induced cytotoxicity.Despite advantages, such as continuous monitoring and more detailed analysis of cytotoxic effects, label-free techniques cannot be generally recommended for cytotoxicity screening of NPs.

View Article: PubMed Central - PubMed

Affiliation: Center for Medical Research, Medical University of Graz, 8010 Graz, Austria.

ABSTRACT
Cytotoxicity testing of nanoparticles (NPs) by conventional screening assays is often complicated by interference. Carbon nanotubes (CNTs) are particularly difficult to assess. To test the suitability of cell-based label-free techniques for this application, a panel of CNTs with different diameters and surface functionalizations was assessed by impedance-based technique (xCELLigence RTCA) and automated microscopy (Cell-IQ) compared to formazan bioreduction (MTS assay). For validation of the label-free systems different concentrations of ethanol and of amine (AMI) polystyrene NPs were used. CNTs were evaluated in various cell lines, but only endothelial EAhy926 cells and L929 and V79 fibroblasts could be evaluated in all systems. Polystyrene particles obtained similar results in all assays. All systems identified thin (<8 nm) CNTs as more cytotoxic than thick (>20 nm) CNTs, but detection by xCELLigence system was less sensitive to CNT-induced cytotoxicity. Despite advantages, such as continuous monitoring and more detailed analysis of cytotoxic effects, label-free techniques cannot be generally recommended for cytotoxicity screening of NPs.

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EAhy926 cells treated for 4 h, 24 h, and 48 h with different concentrations of ethanol (EtOH) and 20 nm amine polystyrene particles (AMI20) assessed by formazan bioreduction (MTS), according to cell index changes in the xCELLigence system (CI), and as stable cells (SC) according to image analysis by Cell-IQ software (n = 3). Changes are normalized to untreated controls as 1 and significant changes are (P < 0.05) indicated by asterisk.
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fig2: EAhy926 cells treated for 4 h, 24 h, and 48 h with different concentrations of ethanol (EtOH) and 20 nm amine polystyrene particles (AMI20) assessed by formazan bioreduction (MTS), according to cell index changes in the xCELLigence system (CI), and as stable cells (SC) according to image analysis by Cell-IQ software (n = 3). Changes are normalized to untreated controls as 1 and significant changes are (P < 0.05) indicated by asterisk.

Mentions: To verify whether all systems identified toxicity of conventional substances or of particles that did not show obvious interference with assay systems, cells were exposed to different concentrations of EtOH and 20 nm AMI particles. While no significant reduction in viability was seen in the MTS, xCELLigence system, and Cell-IQ Analyzer after exposure to 2.5% ethanol, 5% and 10% EtOH reduced viability dramatically (Figure 2).


Suitability of cell-based label-free detection for cytotoxicity screening of carbon nanotubes.

Meindl C, Absenger M, Roblegg E, Fröhlich E - Biomed Res Int (2013)

EAhy926 cells treated for 4 h, 24 h, and 48 h with different concentrations of ethanol (EtOH) and 20 nm amine polystyrene particles (AMI20) assessed by formazan bioreduction (MTS), according to cell index changes in the xCELLigence system (CI), and as stable cells (SC) according to image analysis by Cell-IQ software (n = 3). Changes are normalized to untreated controls as 1 and significant changes are (P < 0.05) indicated by asterisk.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3860081&req=5

fig2: EAhy926 cells treated for 4 h, 24 h, and 48 h with different concentrations of ethanol (EtOH) and 20 nm amine polystyrene particles (AMI20) assessed by formazan bioreduction (MTS), according to cell index changes in the xCELLigence system (CI), and as stable cells (SC) according to image analysis by Cell-IQ software (n = 3). Changes are normalized to untreated controls as 1 and significant changes are (P < 0.05) indicated by asterisk.
Mentions: To verify whether all systems identified toxicity of conventional substances or of particles that did not show obvious interference with assay systems, cells were exposed to different concentrations of EtOH and 20 nm AMI particles. While no significant reduction in viability was seen in the MTS, xCELLigence system, and Cell-IQ Analyzer after exposure to 2.5% ethanol, 5% and 10% EtOH reduced viability dramatically (Figure 2).

Bottom Line: For validation of the label-free systems different concentrations of ethanol and of amine (AMI) polystyrene NPs were used.All systems identified thin (<8 nm) CNTs as more cytotoxic than thick (>20 nm) CNTs, but detection by xCELLigence system was less sensitive to CNT-induced cytotoxicity.Despite advantages, such as continuous monitoring and more detailed analysis of cytotoxic effects, label-free techniques cannot be generally recommended for cytotoxicity screening of NPs.

View Article: PubMed Central - PubMed

Affiliation: Center for Medical Research, Medical University of Graz, 8010 Graz, Austria.

ABSTRACT
Cytotoxicity testing of nanoparticles (NPs) by conventional screening assays is often complicated by interference. Carbon nanotubes (CNTs) are particularly difficult to assess. To test the suitability of cell-based label-free techniques for this application, a panel of CNTs with different diameters and surface functionalizations was assessed by impedance-based technique (xCELLigence RTCA) and automated microscopy (Cell-IQ) compared to formazan bioreduction (MTS assay). For validation of the label-free systems different concentrations of ethanol and of amine (AMI) polystyrene NPs were used. CNTs were evaluated in various cell lines, but only endothelial EAhy926 cells and L929 and V79 fibroblasts could be evaluated in all systems. Polystyrene particles obtained similar results in all assays. All systems identified thin (<8 nm) CNTs as more cytotoxic than thick (>20 nm) CNTs, but detection by xCELLigence system was less sensitive to CNT-induced cytotoxicity. Despite advantages, such as continuous monitoring and more detailed analysis of cytotoxic effects, label-free techniques cannot be generally recommended for cytotoxicity screening of NPs.

Show MeSH