Vascular grafts in canines exhibit similar healing patterns to humans in that the graft surface forms a pseudointima over time but endothelializes only near the anastomotic sites. Thus the pseudointima at the midportion of the graft may represent a nidus for persistent platelet activation. The purpose of this investigation was to examine the effect of the maturing graft surface on platelet activation. Long Dacron subcutaneous carotid to aorta grafts (50 cm x 8 mm) were placed in nine dogs. Blood samples were obtained by direct graft puncture, at the proximal and distal ends of the graft, at 1, 24, 48, 72 hours, 1, 2, 3, 4 weeks, and monthly thereafter for 8 months. Seven sham dogs had subcutaneous grafts implanted without arterial anastomoses, and blood samples were drawn from the femoral artery. Platelet counts were determined with a platelet counter. Platelet aggregation and release of adenosine triphosphate was determined with a whole blood aggregometer by use of arachidonic acid, collagen, and adenosine diphosphate as agonists. No difference was found in platelet aggregation to collagen or adenosine diphosphate stimulation across the graft, but platelets released significantly less adenosine triphosphate to collagen and adenosine diphosphate stimulation distally versus proximally. In the graft dogs a decrease in systemic platelet counts of 50% occurred from the preoperative level which persisted over 8 months (p less than 0.01). Also less response occurred to collagen and ADP stimulated platelet aggregation in the graft animals than the sham animals during the first month of study. These data suggest that significant platelet-graft interactions occur even after the graft has formed a mature pseudointima.