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In vivo cutaneous interferon‐γ gene delivery using novel dicationic (gemini) surfactant–plasmid complexes
Author(s) -
Badea Ildiko,
Verrall Ronald,
BacaEstrada Maria,
Tikoo Suresh,
Rosenberg Alan,
Kumar Praveen,
Foldvari Marianna
Publication year - 2005
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.763
Subject(s) - gene delivery , transfection , chemistry , microbiology and biotechnology , in vivo , liposome , naked dna , genetic enhancement , dna vaccination , plasmid , biology , dna , biochemistry , gene
Abstract Background Localized scleroderma (morphea and linear scleroderma) is a connective tissue disease, accompanied by excessive proliferation and deposition of collagen within the skin, inflammation, vasculopathy and a deranged immune system. Interferon γ (IFNγ), an inhibitor of collagen synthesis and an immunomodulator, could be a potential therapeutic agent if it could be delivered into or expressed locally in affected skin in a non‐invasive manner. In this study, the feasibility of topical delivery of the IFN γ gene and expression of IFNγ were investigated in mice. Methods Novel dicationic (gemini) surfactant (spacer length n = 2–16; alkyl chain m = 12 or 16)–DNA complexes were formulated and characterized by circular dichroism and atomic force microscopy to select gemini analogues with the highest transfection efficiency (TE). Transfection and cellular expression of IFN γ from the bicistronic pGTmCMV.IFN‐GFP plasmid were evaluated in PAM 212 keratinocyte culture by ELISA and fluorescence microscopy. Topical delivery of plasmid using liposomal and nanoemulsion systems, based on gemini surfactant 16‐3‐16, was evaluated in mice by IFNγ expression analysis. Results In vitro TE was found to be dependent on the spacer length of the gemini surfactant, with the C3 spacer showing the highest activity (both 12‐3‐12 and 16‐3‐16). Both gemini cationic liposomes and gemini nanoemulsion (3 × 25 µg DNA/animal) produced significantly higher levels of IFNγ in the skin (359.4 and 607.24 pg/cm 2 ) compared to naked DNA (135.69 pg/cm 2 ) or a liposomal Dc‐chol formulation (82.15 pg/cm 2 ). IFNγ expression in the lymph nodes was higher in the animals treated with gemini liposomes (422.74 pg/animal) compared to the nanoemulsion formulation (131.27 pg/animal) or the Dc‐chol formulation (82pg/animal). Conclusions The feasibility of topical delivery of pGTmCMV.IFN‐GFP plasmid in mice using gemini cationic surfactant based delivery systems was demonstrated. IFNγ expression after treatment with gemini–DNA formulations in the skin was 3–5‐fold higher compared to the treatment with naked DNA ( p < 0.05), and 4–6‐fold higher than the Dc‐chol–DNA complex, indicating a significant advance in topical DNA delivery across intact skin in vivo . Copyright © 2005 John Wiley & Sons, Ltd.