© 2016 The American Association for Thoracic SurgeryBackground The purpose of this study was to provide a preliminary assessment of the performance of a decellularized pericardial patch in a trileaflet aortic valve reconstruction in a long-term juvenile sheep model. Methods A sheep surgical model was used to perform a complete trileaflet reconstruction (Ozaki technique) of the aortic valve with 3 separate pericardial patches. Valve function was assessed 1 week, 3 months, and 6 months after surgery via transthoracic echocardiography. Calcification resistance and host cell infiltration of the pericardial material were assessed at 6 months after surgery. Results Three of 6 sheep with implanted pericardial neo-cusps survived until the planned time of sacrifice after surgery, whereas 3 animals had a successful implant but died shortly after the procedure as the result of a bad recovery from cardiopulmonary bypass. Echocardiography at 6 months revealed a high coaptation area with only minimal regurgitation. In all explanted leaflets, cusp tissue was soft. There was only minimal calcification in 8 of 9 leaflets. Conclusions Aortic valves reconstructed with a decellularized pericardial patch demonstrated adequate diastolic function with minimal regurgitation and resistance to calcification. Combining the Ozaki technique with this decellularized pericardial scaffold showed adequate hemodynamics, sustained mechanical integrity of the patch and limited calcification of the material. These results, together with earlier experimental and clinical data, indicate the potential of this material for aortic valve repair.