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Improved Efficiency of Adenovirus-mediated Vein Grafts Gene Transfer by Use of Poloxamer 407-Trypsin Gel
Wang Xiaowen1,2*, Li Song1, Zhong Changming1, Zhang Cheng1,3, Huang Chun1, Feng Bo2, Lü Zhiqian4
1Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; 2Key Laboratory for Regenerativ Medicine, School of Biomedical Sciences, The Chinese University of Hong Kong, HongKong 999077, China; 3Centre for Clinical Pharmacology, William Harvey Research Institute, Queen Mary University of London, London EC1M 6BQ, United Kingdom; 4Department of Cardiovascular Surgery, Shanghai JiaoTong University Affiliated Sixth People’s Hospital, Shanghai 200233, China
Abstract: Safe and effective vein grafts gene transfer remains elusive in vascular gene therapy. The aim of the present study was to investigate whether poloxamer 407-trypsin gel enhances the efficiency of adenovirus-mediated locally vein grafts gene transfection in vivo. Rat model of vein graft restenosis was established interposition bypass grafting from the autologous jugular vein to the carotid artery. We applied recombinant adenovirus encoding the reporter gene EGFP directly onto vein grafts with poloxamer 407 gel to increase virus contact time， and mild trypsinization to increase virus penetration. The expression of EGFP in vein grafts walls was determined by frozen section， qRT-PCR and immunohistochemistry staining staining at 7， 14 and 28 days after vein graft surgery. Structural integrity of the tissue was evaluated by measurement of tissue tensile strength. Transfection efficiency was significantly higher in vein grafts in poloxamer 407 gel contained varying concentrations of trypsin group versus control group. Trypsin at a concentration of 0.25% allowed marginally better penetration， and tissue tensile strength was not affected. These findings suggest that poloxamer 407-trypsin gel may be a safe and effective method to improve the efficiency of adenovirusmediated vein grafts gene transfer.
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