ABSTRACT
Purpose
There is a lack of relevant, non-animal alternatives for assessing exposure and toxicity of nanoparticle-containing cosmetics, e.g. sunscreens. Our goal was to evaluate timecorrelated single photon counting (TCSPC) for simultaneous monitoring of zinc oxide nanoparticles (ZnO-NP) and the metabolic state of volunteer skin.
Methods
We separated the fluorescence lifetime signatures of endogenous fluorophore signals (i.e. nicotinamide adenine dinucleotide phosphate, NAD(P)H and keratin) and the ZnO-NP signal using advanced TCSPC to simultaneously determine ZnO-NP penetration profiles and NAD(P)H changes in subjects with altered barrier function, including tape-stripped skin and in psoriasis or atopic dermatitis lesions.
Results
We detected no ZnO-NP penetration into viable human skin in any group. ZnO-NP signal was significantly increased (p < 0.01) on the surface of tape-stripped and lesional skin after 4 and 2 h of treatment, respectively. Free NAD(P)H signal significantly increased in tape-stripped viable epidermis treated for 4 h of ZnO-NP compared to vehicle control. No significant NAD(P)H changes were noted in the lesional study.
Conclusion
TCSPC techniques enabled simultaneous, real-time quantification of ZnO-NP concentration and NAD(P)H via non-invasive imaging in the stratum corneum and viable epidermis of volunteers.
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Abbreviations
- AAS:
-
atomic absorption spectroscopy
- AU:
-
arbitrary unit
- CCT:
-
caprylic/capric triglycerides
- FLIM:
-
fluorescence lifetime imaging microscopy
- ICP-OES:
-
inductively coupled plasma-optical emission spectroscopy
- IRF:
-
instrument response function
- KDP:
-
potassium di-hydrogen phosphate
- MEP:
-
multiphoton-excited photoluminescence
- MPT:
-
multiphoton tomography
- MPT-FLIM:
-
multiphoton tomography with fluorescence lifetime imaging microscopy
- NAD(P)H:
-
nicotinamide adenine dinucleotide phosphate
- PBS:
-
phosphate-buffered saline
- SHG:
-
second harmonic generation
- TCSPC:
-
time-correlated single photon counting
- TEM:
-
transmission electron microscope
- TEWL:
-
transepidermal Water Loss
- Ti:Sa:
-
titanium:sapphire
- ZnO-NP:
-
zinc oxide nanoparticles
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ACKNOWLEDGMENTS & DISCLOSURES
We would like to thank the National Health and Medical Research Council of Australia (ID# 569694) and the United States Air Force Asian Office of Aerospace Research and Development for funding. We also thank Corinne Yoong for recruiting volunteers for the lesion studies.
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Lin, L.L., Grice, J.E., Butler, M.K. et al. Time-Correlated Single Photon Counting For Simultaneous Monitoring Of Zinc Oxide Nanoparticles And NAD(P)H In Intact And Barrier-Disrupted Volunteer Skin. Pharm Res 28, 2920–2930 (2011). https://doi.org/10.1007/s11095-011-0515-5
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DOI: https://doi.org/10.1007/s11095-011-0515-5