Spectroscopic analysis confirms the interactions between single walled carbon nanotubes and various dyes commonly used to assess cytotoxicity

Casey, Alan; Herzog, Eva; Davoren, Maria; Lyng, Fiona M.; Byrne, Hugh J.; Chambers, Gary

doi:10.1016/j.carbon.2007.03.033

Cited by 275 publications

(214 citation statements)

References 34 publications

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“…At least three independent experiments were conducted with six replicate wells employed per concentration per plate in each independent experiment. After 24 hours of cell attachment, plates were washed with 100µl/well PBS and treated with increasing concentrations of DOX, incubated for the required exposure time and then the viability was assessed with the aid of the Alamar blue assay as previously reported (Casey et al, 2007 andDavoren et al, 2007). Briefly, the control medium and/or the test exposure was removed, the cells were rinsed with PBS and 100 μl of AB medium (5% [v/v] solution of AB) prepared in fresh medium (without FBS or supplements) were added to each well.…”

Section: Alamar Blue Viability Measurementsmentioning

confidence: 99%

Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices

Casey¹,

Gargotti²,

Bonnier

et al. 2016

Toxicology in Vitro

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The interest in the use of 3D matrices for in vitro analysis, with a view to increasing the relevance of in vitro studies and reducing the dependence on in vivo studies, has been growing in recent years.Cells grown in a 3D in vitro matrix environment have been reported to exhibit significantly different properties to those in a conventional 2D culture environment. However, comparison of 2D and 3D cell culture models have recently been noted to result in differing responses of cytotoxic assays, without any associated change in viability. The effect was attributed to differing conversion rates and effective concentrations of the resazurin assay in 2D and 3D environments, rather than differences in cellular metabolism. In this study, the efficacy of a chemotherapeutic agent, doxorubicin, is monitored and compared in conventional 2D and 3D collagen gel exposures of immortalized human cervical cells. Viability was monitored with the aid of the Alamar Blue assay and drug internalisation was verified using confocal microscopy. Drug uptake and retention within the collagen matrix was monitored by absorption spectroscopy. The viability studies showed apparent differences between the 2D and 3D culture systems, the differences attributed in part to the physical transition from 2D to a 3D environment causing alterations to dye resazurin uptake and conversion rates. The use of 3D culture matrices has widely been interpreted to result in "reduced" toxicity or cellular "resistance" to the chemotherapeutic agent. The results of this study show that the reduced efficiency of the drug to cells grown in the 3D environment can be accounted for by a sequential reduction of the effective concentration of the test compound and assay. This is due to absorption within the collagen gel inducing a higher uptake of both drug and assay thereby influencing the toxic impact of the drug and conversion rate of resazurin, and. the increased effective surface area of the cell exposed to the drug and assay in the 3D environment. The effect was noted to be higher after shorter exposure periods and should be accounted for in in vitro 2D and 3D culture environment comparisons.

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Section: Alamar Blue Viability Measurementsmentioning

confidence: 99%

Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices

Casey¹,

Gargotti²,

Bonnier

et al. 2016

Toxicology in Vitro

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“…The cells were counted at 24-h intervals and a cytometer was used to determine the cell proliferation rate. We did not use the MTT assay or other spectroscopic analyses to determine cell -7 -proliferation because carbon nanotubes interact with these dyes (Casey et al 2007). The cells were stained with trypan blue dye to assess cytotoxicity.…”

Section: Cell Culture and Mwcnt Exposurementioning

confidence: 99%

Proteomics-based safety evaluation of multi-walled carbon nanotubes

Haniu

Matsuda

Takeuchi

et al. 2010

Toxicology and Applied Pharmacology

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“…8 displays the obtained cytotoxic response curve of A549 cells following direct exposure to SWCNT as evaluated by the AB assay. Data of this nature would obviously contain a contribution from the demonstrated indirect effect of medium depletion coupled with absorptive interferences (Casey et al, 2007b;Hurt et al, 2006;Monteiro-Riviere and Inman, 2006;Wörle-Knirsch et al, 2006) from remnant SWCNT after initial exposure both contributing to false positive toxic effects being recorded when evaluating high concentration SWCNT exposures.…”

Section: Cytotoxicity Evaluationmentioning

confidence: 99%

Single walled carbon nanotubes induce indirect cytotoxicity by medium depletion in A549 lung cells

Casey¹,

Herzog²,

Lyng³

et al. 2008

Toxicology Letters

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165

113

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Single walled carbon nanotubes induce indirect cytotoxicity by medium depletion in A549 lung cells." Toxicology Letters, 179, 78-84 (2008) t r a c tThe ability of two types of single walled carbon nanotubes (SWCNT), namely Arc Discharge (AD) and HiPco ® single walled carbon nanotubes, to induce an indirect cytotoxicity in A549 lung cells by means of medium depletion was investigated. The nanotubes were dispersed in a commercial cell culture medium and subsequently removed by centrifugation and filtration. Spectroscopic analysis confirmed the removal of the nanotubes and showed differing degrees of alteration of the composition of the medium upon the removal of the nanotubes. The ability to induce an indirect cytotoxic effect by altering the medium was evaluated using two endpoints, namely the Alamar Blue (AB) and the Clonogenic assay. Exposure of the A549 cells to the depleted medium which had previously contained carbonaceous nanoparticles, revealed significant cytotoxicity for both endpoints employed. The results presented demonstrate that single walled carbon nanotubes can induce an indirect cytotoxicity by alteration of cell culture medium (in which they have previously been dispersed) which potentially results in a false positive toxic effect being observed in cytotoxicity studies.

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Spectroscopic analysis confirms the interactions between single walled carbon nanotubes and various dyes commonly used to assess cytotoxicity

Cited by 275 publications

References 34 publications

Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices

Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices

Proteomics-based safety evaluation of multi-walled carbon nanotubes

Single walled carbon nanotubes induce indirect cytotoxicity by medium depletion in A549 lung cells

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