Experimental analysis for reconsruction of inferior vena cava and portal vein in abdominal surgery : endothelial seeding on artificial grafts and insulin gene transfer into endothelial cells.

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Purpose of the study is to investigate the effectiveness of endothelial seeding on the the artificial graft and insulin gene transfer to endothelial cells on the graft, which was interposed in the canine infreior vena cava and portal vein for vascular reconstruction in hepato-biliary-pancreatic surgery.We could not find any significant difference in patency of the vein grafts, ePTFE grfats, or endothelialized grafts, which were interposed in the IVC or PV.We performed insulin gene transfer using adenovirus vector carrying human insulin gene (AxIns). AxIns was constructed as follows ; human preproinsulin gene genomic DNA (1.3-kb) containing coding sequence, was inserted into the SwaI site of the full-length replication-deficient adenovirus genome cloned in a cassette cosmid. The cosmid was then co-transfected to 293 cells together with the adenovirus DNA-terminal protein complex digested at several sites with EcoT22I.AxIns was generated by overlapping recombination. AxIns was propagated in 293 cells and concentrated with purification using sequential ultracentrifugation in CsCl step gradients. Transfection of AxIns into endothelial cells induced the expression of preproinsulin mRNA and secret proinsulin. One shot injection of AxIns into the spleen transfered human insulin gene mainly in the liver and produced human insulin enough to decrease blood glucose levels in streptozotosin-induced diabetic rats and to restore the serum levels of total protein and albumin, which were impaired in diabetic rats. AxIns injection into the spleen, but not periodic insulin administration, controled blood glucose levels, restored serum total protein levels, and prolonged survival periods after total pacreatectomy in canine models. Thus we confirmed availability of AxIns for in vivo insulin gene transfer.