Tissue engineering mean a wide field of science application in searching the substitusion by different process of damaged human tissue. The main intent is to obtain scaffold that can perfectly mimic the structural and functional characteristics of damaged tissues or organs. In the field of vascular graft numerous studies were conducted to obtain suitable tissue that could supplant the use of autologous bypassor even the littlest vessel employed in microsurgery. Over the years, various solutions have been proposed. The use of resorbable or non-resorbable vascular synthetic conduits, associated or not to stem cells and growth factors, small-caliber decellularized vessels and scaffold-free methods have been described in the literature. First, our study has produced experimental animal models in which a section of the rabbit femoral vessels was replaced by small caliber (inner diameter < 6 mm) human decellularized vessel. Materials and methods Two arteries and two human veins of small caliber (< 6 mm) obtained by upper arm lesions not suitable for surgical procedure were withdrawn and subjected to a process of decellularization according to a modified Sheridan’s protocol. The effectiveness of the process was shown in vitro with histological, immunohistochemical, ultrastructural, and qualitative and quantitative analysis of DNA. Mechanical testing and functional ecodoppler in vivo were also performed. Decellularized vessels were implanted in vivo in rabbit femoral vessels. The patency was verified, in vivo, at time 0, 10 and 20 minutes later. After 30 days for vein implants and 10 and 40 days for artery implants, the rabbits were sacrificed. The patency of grafts, the inflammatory process and re-endothelialization was tested with histological and immunophenotypical analysis. At the same time we analyzed also synthetic vessel as control , after two weeks of implantation. Results The various assays of decellularized vessels, in vitro, demonstrated the success of the process. The cellular component was eliminated without altering the composition and three-dimensional structure of extra cellular matrix. It was also confirmed by mechanical testing. The scaffolds were implanted successfully in all models and the patency was maintained until animals’ sacrifice, as also documented by ecocolordoppler evaluation. The histological and immunophenotypical analysis of taken scaffolds showed a minimal inflammatory process, the maintenance of the extracellular matrix structure and the wall and luminal surface re-cellularization. The syntethic vessel demonstrate complete thrombosis. Conclusions The adjustments made to the decellularization protocol proposed by Sheridan helped to keep acceptable length of the process maintaining the same effectiveness. Non-use of endothelial stem cells reduced costs even if, for longer grafts the use of cells to improve biocompatibility seems to be necessary. The histological studies on withdrawn grafts have demonstrated a minimal presence of inflammatory processes, a complete patent in two weeks till eight weeks and the presence of a process of re-cellularization. The excellent biocompatibility of implanted scaffolds suggest the possibility of a subsequent application in clinical microvascular field and, in the future, for the establishment of a tissue bank to find substitutes with suitable caliber and length. Additional animal testing, with long time survival, mechanical and histological analysis are definitely essential to determine the effectiveness of these implants and think of a possible clinical trials.