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In Vitro Cutaneous Application of ISCOMs on Human Skin Enhances Delivery of Hydrophobic Model Compounds Through the Stratum Corneum

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Abstract

This study aimed to investigate the effect of a novel kind of immune-stimulating complexes (ISCOMs) on human skin penetration of model compounds in vitro to evaluate their potential as a delivery system, ultimately for transcutaneous vaccination. Special focus was on elucidating the mechanisms of penetration. Preparation of ISCOMs was done by dialysis and subsequent purification in a sucrose density gradient. The penetration pathways of acridine-labeled ISCOMs were visualized using confocal laser scanning microscopy (CLSM). Transmission electron microscopy (TEM) was used to evaluate the ultrastructural changes in the skin after application of the ISCOMs with or without hydration. Transcutaneous permeation of the model compound, methyl nicotinate, was evaluated in diffusion cells. The prepared ISCOMs were 42–52 nm in diameter as evaluated by dynamic light scattering with zeta potentials of −33 to −26.1 mV. TEM investigations verified the presence of ISCOM structures. Penetration of acridine into skin was greatly increased by incorporation into ISCOMs as visualized by CLSM. Permeation of methyl nicotinate was enhanced in the presence of ISCOMs. Ultrastructural changes of the intercellular space in the stratum corneum after exposure of ISCOMs were observed on micrographs, especially for hydrated skin. In conclusion, cutaneous application of ISCOMs leads to increased penetration of hydrophobic model compounds through human stratum corneum and thus shows potential as a transcutaneous delivery system. The increased penetration seems to be reflected by a change in the intercellular space between the corneocytes, and the effect is most likely caused by the components of the ISCOMs rather than intact ISCOMs.

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Abbreviations

APC:

Antigen-presenting cells

CLSM:

Confocal laser scanning microscopy

DLS:

Dynamic light scattering

ISCOMs:

Immune-stimulating complexes

LDE:

Laser Doppler electrophoresis

TEM:

Transmission electron microscopy

UC:

Ultracentrifugation

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Acknowledgements

This study was financially supported by the Danish National Advanced Technology Foundation and the Drug Research Academy. Karina Juul Vissing is acknowledged for technical assistance and Niels Coley for linguistic revision of the manuscript. Nordic vaccine A/S is greatly acknowledged for their kind sharing of know-how on the preparation and characterization of the Posintro™ nanoparticles and for their helpful discussions. Furthermore, Nancy Monteiro-Riviere and Al Inman are acknowledged for their kind help on TEM studies with ruthenium tetroxide.

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Authors and Affiliations

  1. Department of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark

    Henriette Baun Madsen, Birger Brodin & Hanne Mørck Nielsen

  2. Coloplast A/S, Bakkegårdsvej 406A, 3050, Humlebæk, Denmark

    Peter Ifversen & Flemming Madsen

  3. Universitäts-Hautklinik, Vossstrasse 2, 69115, Heidelberg, Germany

    Ingrid Hausser

Authors
  1. Henriette Baun Madsen
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  2. Peter Ifversen
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  3. Flemming Madsen
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  4. Birger Brodin
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  5. Ingrid Hausser
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  6. Hanne Mørck Nielsen
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Correspondence to Hanne Mørck Nielsen.

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Madsen, H.B., Ifversen, P., Madsen, F. et al. In Vitro Cutaneous Application of ISCOMs on Human Skin Enhances Delivery of Hydrophobic Model Compounds Through the Stratum Corneum. AAPS J 11, 728–739 (2009). https://doi.org/10.1208/s12248-009-9149-5

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  • DOI: https://doi.org/10.1208/s12248-009-9149-5

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