Treatments of vascular disease via modulating the expression of specific proteins by gene transfer have been attempted in various studies over the past few years. Among several methods to deliver genes, adenovirus currently has been used because of a number of positive aspects. In this study, we test adenoviral vector as a potential mediator in the treatment of vascular disease by using freshly isolated vascular tissues not cultured vascular cells. Freshly isolated vascular tissues were directly exposed to adenoviral vector pAd5CMVmcsIRESeGFPpA to check the possibility of GFP expression in different layer of vascular tissues. We found that the GFP expression by using adenoviral vector experiments is mainly focused on the adventitia and failed to detect GFP expression at endothelial layer or vascular smooth muscle layer in vascular tissues. However, we also found that several integrin receptors are robustly expressed in vascular smooth muscle, thus the limited expression of protein in vascular smooth muscle are not likely the lack of integrin receptors. In conclusion, adenovirus could not be a good tool for a specific protein expression in vascular smooth muscle cell. Thus, the application of adenovirus as a tool for gene therapy of vascular smooth muscle cells in clinical therapeutic trial need to be optimized further.