Cell Biochemistry and Biophysics

, Volume 75, Issue 3–4, pp 359–367 | Cite as

EPR Technology as Sensitive Method for Oxidative Stress Detection in Primary and Secondary Keratinocytes Induced by Two Selected Nanoparticles

  • S. B. Lohan
  • S. Ahlberg
  • A. Mensch
  • D. Höppe
  • M. Giulbudagian
  • M. Calderón
  • S. Grether-Beck
  • J. Krutmann
  • J. Lademann
  • M. C. Meinke
Original Paper


Exogenous factors can cause an imbalance in the redox state of biological systems, promoting the development of oxidative stress, especially reactive oxygen species (ROS). To monitor the intensity of ROS production in secondary keratinocytes (HaCaT) by diesel exhaust particles and thermoresponsive nanogels (tNG), electron paramagnetic resonance (EPR) spectroscopy after 1 and 24 h of incubation, respectively, was applied. Their cytotoxicity was analyzed by a cell viability assay (XTT). For tNG an increase in the cell viability and ROS production of 10% was visible after 24 h, whereas 1 h showed no effect. A ten times lower concentration of diesel exhaust particles exhibited no significant toxic effects on HaCaT cells for both incubation times, thus normal adult human keratinocytes (NHK) were additionally analyzed by XTT and EPR spectroscopy. Here, after 24 h a slight increase of 18% in metabolic activity was observed. However, this effect could not be explained by the ROS formation. A slight increase in the ROS production was only visible after 1 h of incubation time for HaCaT (9%) and NHK (14%).


Oxidative stress Electron paramagnetic resonance spectroscopy Nanoparticles Cell viability 



The authors acknowledge the support of the Deutsche Forschungsgemeinschaft (DFG)/ German Research Foundation via SFB 1112, Projects A04, B01.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • S. B. Lohan
    • 1
    • 2
  • S. Ahlberg
    • 1
    • 2
  • A. Mensch
    • 1
    • 2
  • D. Höppe
    • 1
    • 2
  • M. Giulbudagian
    • 3
  • M. Calderón
    • 3
  • S. Grether-Beck
    • 4
  • J. Krutmann
    • 4
  • J. Lademann
    • 1
    • 2
  • M. C. Meinke
    • 1
    • 2
  1. 1.Charité – Universitätsmedizin Berlin, corporate member of Freie Universität BerlinHumboldt-Universität zu BerlinBerlinGermany
  2. 2.Berlin Institute of Health, Department of Dermatology, Venerology and Allergology, Center of Experimental and Applied Cutaneous PhysiologyCharité—Universitätsmedizin BerlinBerlinGermany
  3. 3.Freie Universität Berlin, Institute of Chemistry and BiochemistryBerlinGermany
  4. 4.Leibniz Research Institute for Environmental MedicineDüsseldorfGermany

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