skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

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]
+ Show Author Affiliations
  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.
Copy to clipboard
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.
Copy to clipboard
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.
Copy to clipboard
@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}
}
Copy to clipboard
Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI:  10.3390/nano9121765
Copyright Statement
Other availability
Search WorldCat Search WorldCat to find libraries that may hold this journal
Save / Share:
Export Metadata
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.

Works referenced in this record:

Binding of blood proteins to carbon nanotubes reduces cytotoxicity
journal, October 2011

  • Ge, C.; Du, J.; Zhao, L.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 41
  • DOI: 10.1073/pnas.1105270108

Mechanisms of lung fibrosis induced by carbon nanotubes: towards an Adverse Outcome Pathway (AOP)
journal, December 2015

  • Vietti, Giulia; Lison, Dominique; van den Brule, Sybille
  • Particle and Fibre Toxicology, Vol. 13, Issue 1
  • DOI: 10.1186/s12989-016-0123-y

Effect of BSA on carbon nanotube dispersion for in vivo and in vitro studies
journal, January 2007

  • Elgrabli, Dan; Abella-Gallart, Steve; Aguerre-Chariol, Olivier
  • Nanotoxicology, Vol. 1, Issue 4
  • DOI: 10.1080/17435390701775136

Advanced computational modeling for in vitro nanomaterial dosimetry
journal, October 2015

  • DeLoid, Glen M.; Cohen, Joel M.; Pyrgiotakis, Georgios
  • Particle and Fibre Toxicology, Vol. 12, Issue 1
  • DOI: 10.1186/s12989-015-0109-1

MWCNTs of different physicochemical properties cause similar inflammatory responses, but differences in transcriptional and histological markers of fibrosis in mouse lungs
journal, April 2015

  • Poulsen, Sarah S.; Saber, Anne T.; Williams, Andrew
  • Toxicology and Applied Pharmacology, Vol. 284, Issue 1
  • DOI: 10.1016/j.taap.2014.12.011

Profibrotic Activity of Multiwalled Carbon Nanotubes Upon Prolonged Exposures in Different Human Lung Cell Types
journal, March 2019

  • Chortarea, Savvina; Zerimariam, Fikad; Barosova, Hana
  • Applied In Vitro Toxicology, Vol. 5, Issue 1
  • DOI: 10.1089/aivt.2017.0033

Endocytosis of Multiwalled Carbon Nanotubes in Bronchial Epithelial and Mesothelial Cells
journal, January 2015

  • Maruyama, Kayo; Haniu, Hisao; Saito, Naoto
  • BioMed Research International, Vol. 2015
  • DOI: 10.1155/2015/793186

In vitro nanoparticle dosimetry for adherent growing cell monolayers covering bottom and lateral walls
journal, October 2018

  • Böhmert, Linda; König, Laura; Sieg, Holger
  • Particle and Fibre Toxicology, Vol. 15, Issue 1
  • DOI: 10.1186/s12989-018-0278-9

Mouse pulmonary dose- and time course-responses induced by exposure to multi-walled carbon nanotubes
journal, March 2010

Use of Coefficient of Variation in Assessing Variability of Quantitative Assays
journal, November 2002

Dispersing Individual Single-Wall Carbon Nanotubes in Aqueous Surfactant Solutions below the cmc
journal, November 2009

  • Angelikopoulos, Panagiotis; Gromov, Andrei; Leen, Ailsa
  • The Journal of Physical Chemistry C, Vol. 114, Issue 1
  • DOI: 10.1021/jp905925r

Distribution and persistence of pleural penetrations by multi-walled carbon nanotubes
journal, January 2010

  • Mercer, Robert R.; Hubbs, Ann F.; Scabilloni, James F.
  • Particle and Fibre Toxicology, Vol. 7, Issue 1
  • DOI: 10.1186/1743-8977-7-28

Interactions and effects of BSA-functionalized single-walled carbon nanotubes on different cell lines
journal, February 2016

Toxicological assessment of multi-walled carbon nanotubes on A549 human lung epithelial cells
journal, March 2015

  • Visalli, Giuseppa; Bertuccio, Maria Paola; Iannazzo, Daniela
  • Toxicology in Vitro, Vol. 29, Issue 2
  • DOI: 10.1016/j.tiv.2014.12.004

Extinction coefficient of water-based multi-walled carbon nanotube nanofluids for application in direct-absorption solar collectors
journal, October 2014

  • Lee, Seung-Hyun; Kim, Hyun Jin; Kim, Kyu Han
  • Micro & Nano Letters, Vol. 9, Issue 10
  • DOI: 10.1049/mnl.2014.0262

Systematic Analysis of Multiwalled Carbon Nanotube-Induced Cellular Signaling and Gene Expression in Human Small Airway Epithelial Cells
journal, February 2013

  • Snyder-Talkington, Brandi N.; Pacurari, Maricica; Dong, Chunlin
  • Toxicological Sciences, Vol. 133, Issue 1
  • DOI: 10.1093/toxsci/kft019

On the pivotal role of dose for particle toxicology and risk assessment: exposure is a poor surrogate for delivered dose
journal, December 2017

Ultrafast Carrier Dynamics in Single-Wall Carbon Nanotubes
journal, February 2003

Oxide Nanoparticle Uptake in Human Lung Fibroblasts:  Effects of Particle Size, Agglomeration, and Diffusion at Low Concentrations
journal, December 2005

  • Limbach, Ludwig K.; Li, Yuchun; Grass, Robert N.
  • Environmental Science & Technology, Vol. 39, Issue 23
  • DOI: 10.1021/es051043o

Revealing the Role of Epithelial Mechanics and Macrophage Clearance during Pulmonary Epithelial Injury Recovery in the Presence of Carbon Nanotubes
journal, October 2018

  • Septiadi, Dedy; Abdussalam, Wildan; Rodriguez-Lorenzo, Laura
  • Advanced Materials, Vol. 30, Issue 52
  • DOI: 10.1002/adma.201806181

Microwave-assisted digestion procedures for biological samples with diluted nitric acid: Identification of reaction products
journal, July 2009

A Simple Spectroscopic Method for Differentiating Cellular Uptakes of Gold Nanospheres and Nanorods from Their Mixtures
journal, February 2010

  • Cho, Eun Chul; Liu, Yang; Xia, Younan
  • Angewandte Chemie International Edition, Vol. 49, Issue 11
  • DOI: 10.1002/anie.200906584

Chemical oxidation of multiwalled carbon nanotubes
journal, May 2008

Quantification of the cellular dose and characterization of nanoparticle transport during in vitro testing
journal, December 2015

  • Rischitor, Grigore; Parracino, Mariantonietta; La Spina, Rita
  • Particle and Fibre Toxicology, Vol. 13, Issue 1
  • DOI: 10.1186/s12989-016-0157-1

Pulmonary fibrotic response to aspiration of multi-walled carbon nanotubes
journal, January 2011

  • Mercer, Robert R.; Hubbs, Ann F.; Scabilloni, James F.
  • Particle and Fibre Toxicology, Vol. 8, Issue 1
  • DOI: 10.1186/1743-8977-8-21

Dispersion Behaviour of Silica Nanoparticles in Biological Media and Its Influence on Cellular Uptake
journal, October 2015

A Multi-Stakeholder Perspective on the Use of Alternative Test Strategies for Nanomaterial Safety Assessment
journal, July 2013

  • Nel, Andre E.; Nasser, Elina; Godwin, Hilary
  • ACS Nano, Vol. 7, Issue 8
  • DOI: 10.1021/nn4037927

Quantitative analysis of the deposited nanoparticle dose on cell cultures by optical absorption spectroscopy
journal, October 2016

  • Spyrogianni, Anastasia; Herrmann, Inge K.; Lucas, Miriam S.
  • Nanomedicine, Vol. 11, Issue 19
  • DOI: 10.2217/nnm-2016-0243

Estimating the effective density of engineered nanomaterials for in vitro dosimetry
journal, March 2014

  • DeLoid, Glen; Cohen, Joel M.; Darrah, Tom
  • Nature Communications, Vol. 5, Issue 1
  • DOI: 10.1038/ncomms4514

ISD3: a particokinetic model for predicting the combined effects of particle sedimentation, diffusion and dissolution on cellular dosimetry for in vitro systems
journal, January 2018

  • Thomas, Dennis G.; Smith, Jordan N.; Thrall, Brian D.
  • Particle and Fibre Toxicology, Vol. 15, Issue 1
  • DOI: 10.1186/s12989-018-0243-7

A review on protein functionalized carbon nanotubes
journal, January 2015

  • Nagaraju, Kathyayini; Reddy, Roopa; Reddy, Narendra
  • Journal of Applied Biomaterials & Functional Materials, Vol. 13, Issue 4
  • DOI: 10.5301/jabfm.5000231

A new angle on dynamic depolarized light scattering: number-averaged size distribution of nanoparticles in focus
journal, January 2016

  • Geers, Christoph; Rodriguez-Lorenzo, Laura; Andreas Urban, Dominic
  • Nanoscale, Vol. 8, Issue 34
  • DOI: 10.1039/C6NR03386E

In vitro approaches to assess the hazard of nanomaterials
journal, October 2017

    [ × clear filter / sort ]

    Similar Records in DOE PAGES and OSTI.GOV collections: