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Pre-clinical evaluation of liposomal gene transfer to improve dermal and epidermal regeneration

Gene Therapy volume 17, pages 770–778 (2010)Cite this article

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Abstract

Liposomal gene transfer effectively enhances dermal and epidermal regeneration in burned rodents. To advance this treatment to clinical studies, we investigated the efficacy of liposomal gene transfer in a clinically relevant porcine wound model. Mimicking the clinical scenario, six female Yorkshire pigs (40–50 kg) received up to 12 burns of 50 cm2 area that were fully excised and covered with skin autograft meshed at 4:1 ratio 24 h post-burn. Animals received control injections (empty liposomes), liposomes (DMRIE-C) containing 1 mg LacZ-cDNA, or liposomes (DMRIE-C) with 1 mg of platelet-derived growth factor (PDGF)-cDNA, or the naked PDGF gene. Serial biopsies were taken from different wound sites at multiple time points up to 12 days post-wounding. Transfection efficacy and transfection rate of LacZ and localization of β-gal were determined by immunohistochemical and immunofluorescent techniques. RT-PCR and multiplex protein analysis (ELISA) were used to measure levels of growth factor mRNA transcribed and growth factor protein translated. Wound re-epithelialization and graft adhesion was evaluated using planimetric analysis and clinical scores. We found that peak transfection of liposomal β-galactosidase occurred on day 2, with a fluorescence increase of 154% to baseline (P<0.001). Transfection intensity dropped to 115% above baseline on day 4 (P<0.001) and 109% on day 7. Immunohistochemistry showed a maximum transfection rate of 34% of cells in wound tissue. Gene transfer of liposomal PDGF-cDNA resulted in increased PDGF-mRNA and protein expression on days 2 and 4, and accelerated wound re-epithlialization as well as graft adhesion on day 9 (P<0.05). In this study, we showed that liposomal cDNA gene transfer is possible in a porcine wound model, and by using PDGF-cDNA we further showed that dermal and epidermal regeneration can be improved. These data indicate that liposomal gene transfer can be a new therapeutic approach to improve wound healing in humans.

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Acknowledgements

We acknowledge the entire staff of the large animal facility of the Shriners Hospitals for Children, especially Thomas Miszalkiewski and John R Salsbury, for their great support in the conduction of the animal studies. We also want to thank Zafar Khakpour, LBI Vienna, for the design and construction of the burn apparatus, and Pat Sellers of the Pathology core laboratory for the preparation of histological slides. We thank Baxter AG, Vienna, Austria, and especially Dr Melitta Bilban, for supporting this study. The work was supported by a Shriners Hospitals for Children (SHC) Research Fellowship Grant no. 8505 and SHC Project/Facility Grants no. 8450, 8460 and 8480. Further support was granted by the Clayton Foundation of Houston.

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Authors and Affiliations

  1. Shriners Hospitals for Children and University of Texas Medical Branch, Galveston, TX, USA

    L K Branski, O E Masters, D N Herndon, D L Traber, R A Cox, K Kita & M G Jeschke

  2. AUVA Research Center—Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Austrian Cluster for Tissue Regeneration, Vienna, Austria

    R Mittermayr & H Redl

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  1. L K Branski
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Correspondence to M G Jeschke.

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Branski, L., Masters, O., Herndon, D. et al. Pre-clinical evaluation of liposomal gene transfer to improve dermal and epidermal regeneration. Gene Ther 17, 770–778 (2010). https://doi.org/10.1038/gt.2010.32

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