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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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Related in: MedlinePlus

Changes in cell index assessed by xCELLigence system in different cell lines after exposure to CNTs normalized to untreated cells as 100% (L929, n = 3; other cell lines n = 2). Significant changes are marked by asterisk and increases by hatch.
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fig5: Changes in cell index assessed by xCELLigence system in different cell lines after exposure to CNTs normalized to untreated cells as 100% (L929, n = 3; other cell lines n = 2). Significant changes are marked by asterisk and increases by hatch.

Mentions: After 4 h, the relative cell index of EAhy926 cells was strongly decreased (<80%) after incubation with SCNT and SCNTc. After 24 h also incubation with MCNT8, MCNT8c, and MCNT50 showed pronounced viability loss (Figure 5). After 24 h, viability of L929 cells was significantly reduced after incubation with MCNT20c. Transient decrease of viability was seen for MCNT20. The cytotoxicity pattern of MRC-5 cells was very similar to that of EAhy926 cells with prominent decreases for SCNT, SCNTc, and MCNT8 and lower cytotoxicity of the remaining CNTs. Differences between 24 h and 48 h of exposure were minimal.


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)

Changes in cell index assessed by xCELLigence system in different cell lines after exposure to CNTs normalized to untreated cells as 100% (L929, n = 3; other cell lines n = 2). Significant changes are marked by asterisk and increases by hatch.
© Copyright Policy
Related In: Results  -  Collection

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

fig5: Changes in cell index assessed by xCELLigence system in different cell lines after exposure to CNTs normalized to untreated cells as 100% (L929, n = 3; other cell lines n = 2). Significant changes are marked by asterisk and increases by hatch.
Mentions: After 4 h, the relative cell index of EAhy926 cells was strongly decreased (<80%) after incubation with SCNT and SCNTc. After 24 h also incubation with MCNT8, MCNT8c, and MCNT50 showed pronounced viability loss (Figure 5). After 24 h, viability of L929 cells was significantly reduced after incubation with MCNT20c. Transient decrease of viability was seen for MCNT20. The cytotoxicity pattern of MRC-5 cells was very similar to that of EAhy926 cells with prominent decreases for SCNT, SCNTc, and MCNT8 and lower cytotoxicity of the remaining CNTs. Differences between 24 h and 48 h of exposure were minimal.

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
Related in: MedlinePlus