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Title: 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];  [2]; ORCiD logo [1]; ORCiD logo [1];  [1];  [1];  [1];  [1];  [3]; ORCiD logo [4];  [5];  [6]; ORCiD logo [1]
  1. Univ. of Fribourg (Switzerland). Adolphe Merkle Inst.
  2. Univ. of Fribourg (Switzerland). Adolphe Merkle Inst.; International Iberian Nanotechnology Lab. (Portugal)
  3. Swansea Univ. Medical School (Wales)
  4. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  5. PETA International Science Consortium Ltd., London (England)
  6. 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. 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, and Rothen-Rutishauser, Barbara. Wed . "Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy". United States. doi: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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