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Investigation of TEMPO partitioning in different skin models as measured by EPR spectroscopy - Insight into the stratum corneum
A. Elpelt, D. Ivanov, A. Nováčková, A. Kováčik, M. Sochorová, S. Saeidpour, C. Teutloff, SB. Lohan, J. Lademann, K. Vávrová, S. Hedtrich, MC. Meinke
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Abdomen MeSH
- Cellular Microenvironment MeSH
- Chromatography, Thin Layer MeSH
- Cyclic N-Oxides chemistry MeSH
- Adult MeSH
- Electron Spin Resonance Spectroscopy MeSH
- Epidermis chemistry MeSH
- Skin chemistry cytology MeSH
- Middle Aged MeSH
- Humans MeSH
- Lipids chemistry MeSH
- Young Adult MeSH
- Swine MeSH
- Breast MeSH
- Aged MeSH
- Spectrophotometry, Infrared MeSH
- Spin Labels MeSH
- Skinfold Thickness MeSH
- Ear, External MeSH
- Animals MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Electron paramagnetic resonance (EPR) spectroscopy represents an established tool to study properties of microenvironments, e.g. to investigate the structure and dynamics of biological and artificial membranes. In this study, the partitioning of the spin probe 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in ex vivo human abdominal and breast skin, ex vivo porcine abdominal and ear skin as well as normal and inflammatory in vitro skin equivalents was investigated by EPR spectroscopy. Furthermore, the stratum corneum (SC) lipid composition (as determined by high-performance thin-layer chromatography), SC lipid chain order (probed by infrared spectroscopy) and the SC thickness (investigated by histology) were determined in the skin models. X-band EPR measurements have shown that TEMPO partitions in the lipophilic and hydrophilic microenvironment in varying ratios in different ex vivo and in vitro skin models. Ex vivo human abdominal skin exhibited the highest amount of TEMPO in the lipophilic microenvironment. In contrast, the lowest amount of TEMPO in the lipophilic microenvironment was determined in ex vivo human breast skin and the inflammatory in vitro skin equivalents. Individual EPR spectra of epidermis including SC and dermis indicated that the lipophilic microenvironment of TEMPO mainly corresponds to the most lipophilic part of the epidermis, the SC. The amount of TEMPO in the lipophilic microenvironment was independent of the SC lipid composition and the SC lipid chain order but correlated with the SC thickness. In conclusion, EPR spectroscopy could be a novel technique to determine differences in the SC thickness, thus suitably complementing existing methods.
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- $a Elpelt, Anja $u Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Department of Dermatology, Venerology and Allergology, Center of Experimental and Applied Cutaneous Physiology, Charitéplatz 1, 10117 Berlin, Germany; Institute of Pharmacy, Freie Universität Berlin, Königin-Luise-Straße 2+4, 14195 Berlin, Germany
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- $a Electron paramagnetic resonance (EPR) spectroscopy represents an established tool to study properties of microenvironments, e.g. to investigate the structure and dynamics of biological and artificial membranes. In this study, the partitioning of the spin probe 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in ex vivo human abdominal and breast skin, ex vivo porcine abdominal and ear skin as well as normal and inflammatory in vitro skin equivalents was investigated by EPR spectroscopy. Furthermore, the stratum corneum (SC) lipid composition (as determined by high-performance thin-layer chromatography), SC lipid chain order (probed by infrared spectroscopy) and the SC thickness (investigated by histology) were determined in the skin models. X-band EPR measurements have shown that TEMPO partitions in the lipophilic and hydrophilic microenvironment in varying ratios in different ex vivo and in vitro skin models. Ex vivo human abdominal skin exhibited the highest amount of TEMPO in the lipophilic microenvironment. In contrast, the lowest amount of TEMPO in the lipophilic microenvironment was determined in ex vivo human breast skin and the inflammatory in vitro skin equivalents. Individual EPR spectra of epidermis including SC and dermis indicated that the lipophilic microenvironment of TEMPO mainly corresponds to the most lipophilic part of the epidermis, the SC. The amount of TEMPO in the lipophilic microenvironment was independent of the SC lipid composition and the SC lipid chain order but correlated with the SC thickness. In conclusion, EPR spectroscopy could be a novel technique to determine differences in the SC thickness, thus suitably complementing existing methods.
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