Basic Research
Copyright ©2006 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Oct 14, 2006; 12(38): 6149-6155
Published online Oct 14, 2006. doi: 10.3748/wjg.v12.i38.6149
Hydrodynamics based transfection in normal and fibrotic rats
Rita Yeikilis, Shunit Gal, Natalia Kopeiko, Melia Paizi, Mark Pines, Filip Braet, Gadi Spira
Rita Yeikilis, Shunit Gal, Melia Paizi, Gadi Spira, Department of Anatomy and Cell Biology, The Bruce Rappaport Faculty of Medicine, Rappaport Family Institute for Research in the Medical Sciences, Technion, Haifa 31096, Israel
Natalia Kopeiko, Department of Experimental Surgery, The Bruce Rappaport Faculty of Medicine, Rappaport Family Institute for Research in the Medical Sciences, Technion, Haifa 31096, Israel
Mark Pines, Institute of Animal Sciences, Agricultural Research Organization, the Volcani Center, Bet Dagan 50250, Israel
Filip Braet, Australian Key Centre for Microscopy and Microanalysis, The University of Sydney, Sydney, NSW 2006, Australia
Supported by the Israel Science Foundation, No. 537/01, the C. Rosenblantt Cancer Research Fund and the Rappaport Institute Fund
Correspondence to: Gadi Spira, PhD, Department of Anatomy and Cell Biology, Faculty of Medicine, POB 9649, Haifa 31096, Israel. spira@tx.technion.ac.il
Telephone: +972-4-8295355 Fax: +972-4-8515113
Received: April 20, 2006
Revised: April 28, 2006
Accepted: May 22, 2006
Published online: October 14, 2006
Abstract

AIM: Hydrodynamics based transfection (HBT), the injection of a large volume of naked plasmid DNA in a short time is a relatively simple, efficient and safe method for in vivo transfection of liver cells. Though used for quite some time, the mechanism of gene transfection has not yet been elucidated.

METHODS: A luciferase encoding plasmid was injected using the hydrodynamics based procedure into normal and thioacetamide-induced fibrotic Sprague Dawley rats. Scanning and transmission electron microscopy images were taken. The consequence of a dual injection of Ringer solution and luciferase pDNA was followed. Halofuginone, an anti collagen type I inhibitor was used to reduce ECM load in fibrotic rats prior to the hydrodynamic injection.

RESULTS: Large endothelial gaps formed as soon as 10’ following hydrodynamic injection; these gradually returned to normal 10 d post injection. Hydrodynamic administration of Ringer 10 or 30 m prior to moderate injection of plasmid did not result in efficient transfection suggesting that endothelial gaps by themselves are not sufficient for gene expression. Gene transfection following hydrodynamic injection in thioacetamide induced fibrotic rats was diminished coinciding with the level of fibrosis. Halofuginone, a specific collagen typeIinhibitor, alleviated this effect.

CONCLUSION: The hydrodynamic pressure formed following HBT results in the formation of large endothelial gaps. These gaps, though important in the transfer of DNA molecules from the blood to the space of Disse are not enough to provide the appropriate conditions for hepatocyte transfection. Hydrodynamics based injection is applicable in fibrotic rats provided that ECM load is reduced.

Keywords: Gene transfection, Fibrosis, In vivo transfection, Fibrotic, Hepatic, Endothelial lining, Endothelium, Sinusoidal, Fenestrae, Space of disse, Extracellular matrix