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127. Kongress der Deutschen Gesellschaft für Chirurgie

Deutsche Gesellschaft für Chirurgie

20.04. - 23.04.2010, Berlin

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Non-viral gene therapy for regeneration in an ischemic flap model

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  • Alex Slobodianski - UK-SH, Plastische Chirurgie, Lübeck, Deutschland
  • Christoph Hartog - UK-SH, Plastische Chirurgie, Lübeck, Deutschland
  • Astrid Kathöfer - UK-SH, Plastische Chirurgie, Lübeck, Deutschland
  • Jessica Frenz - Universitätsklinikum Schleswig-Holstein, Campus Lübeck, Plastische Chirurgie, Lübeck, Deutschland
  • Ziyang Zhang - Klinikum rechts der Isar, Klinik für Gefäßchirurgie, München, Deutschland
  • Peter Mailänder - Universitätsklinikum Schleswig-Holstein, Campus Lübeck, Plastische Chirurgie, Handchirurgie, Intensiveinheit für Schwerbrandverletzte, Lübeck, Deutschland
  • Hans-Günther Machens - Klinikum rechts der Isar, Technische Universität München, Klinik und Poliklinik für Plastische Chirurgie und Handchirurgie, München, Deutschland

Deutsche Gesellschaft für Chirurgie. 127. Kongress der Deutschen Gesellschaft für Chirurgie. Berlin, 20.-23.04.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. Doc10dgch639

doi: 10.3205/10dgch639, urn:nbn:de:0183-10dgch6393

Published: May 17, 2010

© 2010 Slobodianski et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

Outline

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Introduction: We established a cell-based, non viral gene-transfer method using fibroblasts to temporarily produce bFGF and VEGF165 in ischemic tissue. Protein delivery from transfected cells can induce expression of tissue inductive factors to stimulate the cellular processes required for regeneration. In the present work, we investigated effects of temporary expression of the angiogenic proteins basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF165) on ischemic tissue for therapeutic purposes.

Materials and methods: The eukaryotic expression vectors harboring VEGF and bFGF cDNAs were transfected into rat primary skin fibroblasts mediated by Amaxa Nucleofector. To determine an improvement in ischemically challenged tissue, a genetically modified cellspool was injected in an ischemic flap model. Gene expression and protein production in vivo and in vitro were measured by real time PCR and immunoassay (BioPlex) respectively. Clinical outcome was demonstrated by planimetric measurements.

Results: Temporary protein expression of bFGF and VEGF165 in the target tissue of the ischemic flap model increased compared to controls after injection of genetically modified cells. A significant improvement of tissue survival was observed after the transfected cell administration. A reduction in flap necrosis by one-third or more was detected after two weeks if transfected cells were applied 1 week before ischemia.

Conclusion: In our work we showed that temporary expression of bFGF and VEGF165 induces therapeutically relevant effects in the rat flap model of ischemia. Standardized high efficiency non viral bFGF and VEGF165 transfection technology can be used in preclinical research.