Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy
Abstract
The overt hazard of carbon nanotubes (CNTs) is often assessed using in vitro methods, but determining a dose–response relationship is still a challenge due to the analytical difficulty of quantifying the dose delivered to cells. An approach to accurately quantify CNT doses for submerged in vitro adherent cell culture systems using UV-VIS-near-infrared (NIR) spectroscopy is provided here. Two types of multi-walled CNTs (MWCNTs), Mitsui-7 and Nanocyl, which are dispersed in protein rich cell culture media, are studied as tested materials. Post 48 h of CNT incubation, the cellular fractions are subjected to microwave-assisted acid digestion/oxidation treatment, which eliminates biological matrix interference and improves CNT colloidal stability. The retrieved oxidized CNTs are analyzed and quantified using UV-VIS-NIR spectroscopy. In vitro imaging and quantification data in the presence of human lung epithelial cells (A549) confirm that up to 85% of Mitsui-7 and 48% for Nanocyl sediment interact (either through internalization or adherence) with cells during the 48 h of incubation. This finding is further confirmed using a sedimentation approach to estimate the delivered dose by measuring the depletion profile of the CNTs.
- Authors:
-
[1];
[1];
[4];
[1]
- Univ. of Fribourg (Switzerland). Adolphe Merkle Inst.
- Univ. of Fribourg (Switzerland). Adolphe Merkle Inst.; International Iberian Nanotechnology Lab. (Portugal)
- Swansea Univ. Medical School (Wales)
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- PETA International Science Consortium Ltd., London (England)
- Univ. of Fribourg (Switzerland). Dept. of Chemistry
- Publication Date:
- Research Org.:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1601354
- Report Number(s):
- PNNL-SA-150718
Journal ID: ISSN 2079-4991; NANOKO
- Grant/Contract Number:
- AC05-76RL01830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nanomaterials
- Additional Journal Information:
- Journal Volume: 9; Journal Issue: 12; Journal ID: ISSN 2079-4991
- Publisher:
- MDPI
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; adherent cell cultures; carbon nanotubes; depletion; quantification; sedimentation; UV-VIS-NIR spectroscopy
Citation Formats
Septiadi, Dedy, Rodriguez-Lorenzo, Laura, Balog, Sandor, Spuch-Calvar, Miguel, Spiaggia, Giovanni, Taladriz-Blanco, Patricia, Barosova, Hana, Chortarea, Savvina, Clift, Martin J. D., Teeguarden, Justin, Sharma, Monita, Petri-Fink, Alke, and Rothen-Rutishauser, Barbara. Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy. United States: N. p., 2019.
Web. doi:10.3390/nano9121765.
Septiadi, Dedy, Rodriguez-Lorenzo, Laura, Balog, Sandor, Spuch-Calvar, Miguel, Spiaggia, Giovanni, Taladriz-Blanco, Patricia, Barosova, Hana, Chortarea, Savvina, Clift, Martin J. D., Teeguarden, Justin, Sharma, Monita, Petri-Fink, Alke, & Rothen-Rutishauser, Barbara. Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy. United States. https://doi.org/10.3390/nano9121765
Septiadi, Dedy, Rodriguez-Lorenzo, Laura, Balog, Sandor, Spuch-Calvar, Miguel, Spiaggia, Giovanni, Taladriz-Blanco, Patricia, Barosova, Hana, Chortarea, Savvina, Clift, Martin J. D., Teeguarden, Justin, Sharma, Monita, Petri-Fink, Alke, and Rothen-Rutishauser, Barbara. Wed .
"Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy". United States. https://doi.org/10.3390/nano9121765. https://www.osti.gov/servlets/purl/1601354.
@article{osti_1601354,
title = {Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy},
author = {Septiadi, Dedy and Rodriguez-Lorenzo, Laura and Balog, Sandor and Spuch-Calvar, Miguel and Spiaggia, Giovanni and Taladriz-Blanco, Patricia and Barosova, Hana and Chortarea, Savvina and Clift, Martin J. D. and Teeguarden, Justin and Sharma, Monita and Petri-Fink, Alke and Rothen-Rutishauser, Barbara},
abstractNote = {The overt hazard of carbon nanotubes (CNTs) is often assessed using in vitro methods, but determining a dose–response relationship is still a challenge due to the analytical difficulty of quantifying the dose delivered to cells. An approach to accurately quantify CNT doses for submerged in vitro adherent cell culture systems using UV-VIS-near-infrared (NIR) spectroscopy is provided here. Two types of multi-walled CNTs (MWCNTs), Mitsui-7 and Nanocyl, which are dispersed in protein rich cell culture media, are studied as tested materials. Post 48 h of CNT incubation, the cellular fractions are subjected to microwave-assisted acid digestion/oxidation treatment, which eliminates biological matrix interference and improves CNT colloidal stability. The retrieved oxidized CNTs are analyzed and quantified using UV-VIS-NIR spectroscopy. In vitro imaging and quantification data in the presence of human lung epithelial cells (A549) confirm that up to 85% of Mitsui-7 and 48% for Nanocyl sediment interact (either through internalization or adherence) with cells during the 48 h of incubation. This finding is further confirmed using a sedimentation approach to estimate the delivered dose by measuring the depletion profile of the CNTs.},
doi = {10.3390/nano9121765},
journal = {Nanomaterials},
number = 12,
volume = 9,
place = {United States},
year = {2019},
month = {12}
}
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