Advertisement

Pharmaceutical Research

, 28:2920 | Cite as

Time-Correlated Single Photon Counting For Simultaneous Monitoring Of Zinc Oxide Nanoparticles And NAD(P)H In Intact And Barrier-Disrupted Volunteer Skin

  • Lynlee L. Lin
  • Jeffrey E. Grice
  • Margaret K. Butler
  • Andrei V. Zvyagin
  • Wolfgang Becker
  • Thomas A. Robertson
  • H. Peter Soyer
  • Michael S. Roberts
  • Tarl W. Prow
Research Paper

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.

KEY WORDS

human skin metabolism multiphoton microscopy sunscreen zinc oxide nanoparticle 

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

Notes

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.

REFERENCES

  1. 1.
    DIRECTIVE 2003/15/EC OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL. Official Journal of the European Union 2003;46:26–35, .Google Scholar
  2. 2.
    Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products Official Journal of the European Union. 52:59–209 (2009).Google Scholar
  3. 3.
    Ruet Rossignol M. The 7th Amendment to the Cosmetics Directive. Altern Lab Anim. 2005;33 Suppl 1:19–20.PubMedGoogle Scholar
  4. 4.
    Robertson TA, Sanchez WY, Roberts MS. Are commercially available nanoparticles safe when applied to the skin? J Biomed Nanotechnol. 2010;6:452–68.PubMedCrossRefGoogle Scholar
  5. 5.
    Ahmed AH, Soyer HP, Saunders N, Boukamp P, Roberts MS. Non-melanoma skin cancers. Drug Discovery Today: Disease Mechanisms: Skin diseases. 2008;5:e55–62.CrossRefGoogle Scholar
  6. 6.
    Leiterand U, Garbe C. Epidemiology of melanoma and nonmelanoma skin cancer–the role of sunlight. Adv Exp Med Biol. 2008;624:89–103.CrossRefGoogle Scholar
  7. 7.
    Monteiro-Riviereand NA, Riviere JE. Interactions of nanomaterials with skin: Aspects of absorption and biodistribution. Nanotoxicology. 2009;3:188–95.CrossRefGoogle Scholar
  8. 8.
    Nohynek GJ, Lademann J, Ribaud C, Roberts MS. Grey goo on the skin? Nanotechnology, cosmetic and sunscreen safety. Crit Rev Toxicol. 2007;37:251–77.PubMedCrossRefGoogle Scholar
  9. 9.
    Singhand S, Nalwa HS. Nanotechnology and health safety–toxicity and risk assessments of nanostructured materials on human health. J Nanosci Nanotechnol. 2007;7:3048–70.CrossRefGoogle Scholar
  10. 10.
    Sternand ST, McNeil SE. Nanotechnology safety concerns revisited. Toxicol Sci. 2008;101:4–21.CrossRefGoogle Scholar
  11. 11.
    Cross SE, Innes B, Roberts MS, Tsuzuki T, Robertson TA, McCormick P. Human skin penetration of sunscreen nanoparticles: In-vitro assessment of a novel micronized zinc oxide formulation. Skin Pharmacol Phys. 2007;20:148–54.CrossRefGoogle Scholar
  12. 12.
    Durand L, Habran N, Henschel V, Amighi K. In vitro evaluation of the cutaneous penetration of sprayable sunscreen emulsions with high concentrations of UV filters. Int J Cosmet Sci. 2009;31:279–92.PubMedCrossRefGoogle Scholar
  13. 13.
    Newman MD, Stotland M, Ellis JI. The safety of nanosized particles in titanium dioxide- and zinc oxide-based sunscreens. J Am Acad Dermatol. 2009;61:685–92.PubMedCrossRefGoogle Scholar
  14. 14.
    Roberts MS, Roberts MJ, Robertson TA, Sanchez W, Thorling C, Zou Y, et al. In vitro and in vivo imaging of xenobiotic transport in human skin and in the rat liver. J Biophotonics. 2008;1:478–93.PubMedCrossRefGoogle Scholar
  15. 15.
    Kortingand HC, Schafer-Korting M. Carriers in the topical treatment of skin disease. Handb Exp Pharmacol :435–468 (2010).Google Scholar
  16. 16.
    Prow TW, Grice JE, Lin LL, Faye R, Butler MK, Becker W, Wurme EMT, Yoong Y, Robertsona TA, Soyer HP, Roberts MS. Nanoparticles and Microparticles for Skin Drug Delivery. Adv Drug Del Rev:In press. (2011).Google Scholar
  17. 17.
    Schneider M, Stracke F, Hansen S, Schaefer UF. Nanoparticles and their interactions with the dermal barrier. Dermatoendocrinol. 2009;1:197–206.PubMedCrossRefGoogle Scholar
  18. 18.
    Zhangand LW, Monteiro-Riviere NA. Assessment of quantum dot penetration into intact, tape-stripped, abraded and flexed rat skin. Skin Pharmacol Phys. 2008;21:166–80.CrossRefGoogle Scholar
  19. 19.
    Samberg ME, Oldenburg SJ, Monteiro-Riviere NA. Evaluation of silver nanoparticle toxicity in skin in vivo and keratinocytes in vitro. Environ Health Perspect. 2010;118:407–13.PubMedCrossRefGoogle Scholar
  20. 20.
    Kuo TR, Wu CL, Hsu CT, Lo W, Chiang SJ, Lin SJ, et al. Chemical enhancer induced changes in the mechanisms of transdermal delivery of zinc oxide nanoparticles. Biomaterials. 2009;30:3002–8.PubMedCrossRefGoogle Scholar
  21. 21.
    Bian SW, Mudunkotuwa IA, Rupasinghe T, Grassian VH. Aggregation and Dissolution of 4 nm ZnO Nanoparticles in Aqueous Environments: Influence of pH, Ionic Strength, Size, and Adsorption of Humic Acid. Langmuir (2011).Google Scholar
  22. 22.
    Gamer AO, Leibold E, van Ravenzwaay B. The in vitro absorption of microfine zinc oxide and titanium dioxide through porcine skin. Toxicology in Vitro. 2006;20:301–7.PubMedCrossRefGoogle Scholar
  23. 23.
    Gulson B, McCall M, Korsch M, Gomez L, Casey P, Oytam Y, et al. Small amounts of zinc from zinc oxide particles in sunscreens applied outdoors are absorbed through human skin. Toxicol Sci. 2010;118:140–9.PubMedCrossRefGoogle Scholar
  24. 24.
    Zvyagin AV, Zhao X, Gierden A, Sanchez W, Ross JA, Roberts MS. Imaging of zinc oxide nanoparticle penetration in human skin in vitro and in vivo. J Biomed Opt. 2008;13:064031.PubMedCrossRefGoogle Scholar
  25. 25.
    Sanchez WY, Prow TW, Sanchez WH, Grice JE, Roberts MS. Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy. J Biomed Opt. 2010;15:046008.PubMedCrossRefGoogle Scholar
  26. 26.
    Ying W. NAD+/NADH and NADP+/NADPH in cellular functions and cell death: regulation and biological consequences. Antioxid Redox Signal. 2008;10:179–206.PubMedCrossRefGoogle Scholar
  27. 27.
    Becker W. Advanced time-correlated single photon counting techniques. Berlin: Springer; 2005.CrossRefGoogle Scholar
  28. 28.
    Berg JM, Romoser A, Banerjee N, Zebda R, Sayes CM. The relationship between pH and zeta potential of similar to 30 nm metal oxide nanoparticle suspensions relevant to in vitro toxicological evaluations. Nanotoxicology. 2009;3:276–83.CrossRefGoogle Scholar
  29. 29.
    Prow TW, Monteiro-Riviere NA, Inman AO, Grice JE, Chen X, Zhao X, Sanchez WH, Gierden A, Kendall MA, Zvyagin AV, Erdmann D, Riviere JE, Roberts MS. Quantum dot penetration into viable human skin. Nanotoxicology 2011.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Lynlee L. Lin
    • 1
    • 2
  • Jeffrey E. Grice
    • 1
  • Margaret K. Butler
    • 3
  • Andrei V. Zvyagin
    • 4
    • 5
  • Wolfgang Becker
    • 6
  • Thomas A. Robertson
    • 7
  • H. Peter Soyer
    • 2
  • Michael S. Roberts
    • 1
    • 7
  • Tarl W. Prow
    • 1
    • 2
    • 8
  1. 1.Therapeutics Research Centre, School of Medicine Princess Alexandra HospitalUniversity of QueenslandBrisbaneAustralia
  2. 2.Dermatology Research Centre, School of Medicine Princess Alexandra HospitalUniversity of QueenslandBrisbaneAustralia
  3. 3.Australian Institute for Bioengineering & NanotechnologyUniversity of QueenslandBrisbaneAustralia
  4. 4.Department of Physics, Centre of MQ PhotonicsMacquarie UniversitySydneyAustralia
  5. 5.Centre for Biophotonics and Laser Science School of Physical SciencesUniversity of QueenslandBrisbaneAustralia
  6. 6.Becker & Hickl GmbHBerlinGermany
  7. 7.Therapeutics Research Centre School of Pharmacy & Biomedical SciencesUniversity of South AustraliaAdelaideAustralia
  8. 8.Therapeutics Research & Dermatology Research Centres School of Medicine, Princess Alexandra HospitalUniversity of QueenslandWoolloongabbaAustralia

Personalised recommendations