EPR Technology as Sensitive Method for Oxidative Stress Detection in Primary and Secondary Keratinocytes Induced by Two Selected Nanoparticles
- 266 Downloads
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%).
KeywordsOxidative 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.
- 10.Delaney, C. A., Green, I. C., Lowe, J. E., Cunningham, J. M., Butler, A. R., Renton, L., et al. (1997). Use of the comet assay to investigate possible interactions of nitric oxide and reactive oxygen species in the induction of DNA damage and inhibition of function in an insulin-secreting cell line. Mutation Research, 375, 137–146.CrossRefPubMedGoogle Scholar
- 11.Fang, L., Neutzner, A., Turtschi, S., Flammer, J., & Mozaffarieh, M. (2015). Comet assay as an indirect measure of systemic oxidative stress. Journal of Visualized Experiments, 99, e52763.Google Scholar
- 15.Mrakic-Sposta, S., Gussoni, M., Montorsi, M., Porcelli, S., & Vezzoli, A. (2014). A quantitative method to monitor reactive oxygen species production by electron paramagnetic resonance in physiological and pathological conditions. Oxidative Medicine and Cellular Longevity, 2014, 306179.CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Lohan, S. B., Bauersachs, S., Ahlberg, S., Baisaeng, N., Keck, C. M., Muller, R. H., et al. (2015). Ultra-small lipid nanoparticles promote the penetration of coenzyme Q10 in skin cells and counteract oxidative stress. European Journal of Pharmaceutics and Biopharmaceutics, 89, 201–207.CrossRefPubMedGoogle Scholar
- 18.Lauer, A. C., Groth, N., Haag, S. F., Darvin, M. E., Lademann, J., & Meinke, M. C. (2013). Radical scavenging capacity in human skin before and after vitamin C uptake: An in vivo feasibility study using electron paramagnetic resonance spectroscopy. The Journal of Investigative Dermatology, 133, 1102–1104.CrossRefPubMedGoogle Scholar
- 22.Lai, P., Lechtman, E., Mashouf, S., Pignol, J. P., & Reilly, R. M. (2016). Depot system for controlled release of gold nanoparticles with precise intratumoral placement by permanent brachytherapy seed implantation (PSI) techniques. International Journal of Pharmaceutics, 515, 729–739.CrossRefPubMedGoogle Scholar
- 25.Rancan, F., Giulbudagian, M., Jurisch, J., Blume-Peytavi, U., Calderon, M., & Vogt, A. (2016). Drug delivery across intact and disrupted skin barrier: Identification of cell populations interacting with penetrated thermoresponsive nanogels. European Journal of Pharmaceutics and Biopharmaceutics, 116, 4–11.Google Scholar
- 31.Ahlberg, S., Meinke, M. C., Werner, L., Epple, M., Diendorf, J., Blume-Peytavi, U., et al. (2014). Comparison of silver nanoparticles stored under air or argon with respect to the induction of intracellular free radicals and toxic effects toward keratinocytes. European Journal of Pharmaceutics and Biopharmaceutics, 88, 651–657.CrossRefPubMedGoogle Scholar
- 33.Riss, T. L., Moravec, R. A., Niles, A. L., Duellman, S., Benink, H. A., Worzella, T. J., et al. (2004). Cell viability assays. In G. S. Sittampalam, N. P. Coussens, H. Nelson, M. Arkin, D. Auld, C. Austin, et al. (Eds.) Assay guidance manual. The National Center for Advancing Translational Sciences (NCATS), Bethesda, MD.Google Scholar
- 35.Gerecke, C., Edlich, A., Giulbudagian, M., Schumacher, F., Zhang, N., Said, A., et al. (2017). Biocompatibility and characterization of polyglycerol-based thermoresponsive nanogels designed as novel drug delivery systems and their intracellular fate in keratinocytes. Nanotoxicology, Under review.Google Scholar
- 40.Fiorito, S., Mastrofrancesco, A., Cardinali, G., Rosato, E., Salsano, F., Sheng Su, D. et al. (2011). Effects of carbonaceous nanoparticles from low-emission and older diesel engines on human skin cells. Carbon, 49, 1–9.Google Scholar