ObjectiveTo evaluate the effect of human heme oxygenase 1 augmentation in vein grafts by adenoviral mediated gene transfer of heme oxygenase 1 (Ad hHO 1) on intimal hyperplasia.MethodsTwenty one Japanese white rabbits were divided into three groups: control group, Ad null control group, and Ad hHO 1 group(each group 7 rabbits). During the operation of rabbits jugular vein into carotid artery interposition grafting, harvested rabbit jugular vein segments were exposed for 30min at room temperature to heparin saline, recombinant replication deficient adenovirus encoding hHO 1(Ad hHO 1, 1× 10 9pfu/ml), and nude recombinant replication deficient adenovirus (Ad null, 1×10 9pfu/ml). Quantitative histological studies of the vein segments were performed 28 days after operation. Protein of hHO 1 was detected with method of immunohistochemical staining(S P) in 14 days and 28 days after operation.ResultsThe average intimal thickness, medial thickness and intimal to medial(I/M) ratio were calculated for each group 28 days after bypass operation. Compared to intimal thickness, I/M ratio of control group veins and Ad null group veins,Ad hHO 1 group veins decreased significantly( P lt;0.01). There was no statistically difference in medial thickness ( P gt;0 05). Strong staining of hHO 1 was detected in vein grafts wall of Ad hHO 1 group.ConclusionAd hHO 1 gene therapy may inhibit intimal hyperplasia of vein grafts in rabbits.
Objective To study the adenovirus-mediated human bone morphogenetic protein-2 gene （Ad-hBMP-2）transferred to the intervertebral disc cells of the New Zealand rabbit in vitro. Methods The cells of New Zealand white rabbitswere isolated from their lumbar discs. The cells were grown in the monolayer and treated with an adenovirus encoding the LacZ gene (Ad-LacZ) and Ad-hBMP-2 (50,100, 150 MOI,multiplicity of infection) in the Dulbecco’s Modified Eagle Medium and the Ham’s F-12 Medium in vitro. Three days after the Ad-hBMP-2 treatment,the expression of hBMP-2 in the cells that had been infected by different dosesof MOI was determined by immunofluorescence and the Western blot analysis, and the expression was determined in the cells with the Ad-LacZ treatment in a dose of 150 MOI. Six days after the Ad-hBMP-2 treatment, mRNA was extracted for the reverse transcription polymerase chain reaction (RT-PCR) and the difference was detected between the control group and the culture group that was treated withAd-hBMP-2 in doses of 50, 100 and 150 MOI so that the expressions of aggrecan and collagen ⅡmRNA could be observed. Results The expression of hBMP-2 in the cells was gradually increased after the transfection in an increasing dose, which was observed by immunofluorescence and the Western blot analysis. At 6 days the aggrecan and collagen type Ⅱ mRNA expressions were up-regulated by Ad-hBMP-2 after the transfection at an increasing viral concentration in the dosedependent manner. Conclusion The results show that Ad-hBMP-2 can transfect the rabbit intervertebral disc cells in vitro with a high efficiency rate and the expression of hBMP-2 after theinfection is dose-dependent in the manner. AdhBMP-2 after transfection can up-regulate the expression of aggrecan and collagen Ⅱ mRNA at an increasing viral concentration.