There is still controversy about the correlation of thrombocytosis and thrombosis complication. Using a rodent splenectomy-induced thrombocytosis model and a thrombogenic endothelial damage model (inverted suture resulting in an intraluminal thrombogenic adventitia of divided femoral artery), the authors investigated whether reactive thrombocytosis with or without endothelial damage contributes to the patency of microvascular anastomosis. Four experimental groups were evaluated in this study: 1) sham operation without thrombogenic anastomosis after femoral artery division; 2) sham operation with thrombogenic anastomosis; 3) thrombocytosis alone without thrombogenic anastomosis; 4) thrombocytosis with thrombogenic anastomosis (each subgroup n = 10, total N = 40). Vascular patency was assessed after immediate operation and on the seventh day postoperatively. Platelet counts and platelet activation (CD62P) were studied in correlation to microvascular patency. In rats without thrombogenic anastomosis groups, there were no significant differences in CD62P expression on platelets (p= 0.09), the patency rates (p= 0.561), or perfusion units (p= 0.746) before and after arterial reanastomosis between rats with and without thrombocytosis, respectively. However, the thrombogenic anastomosis of femoral artery in thrombocytosis and control groups showed significantly increased CD62P expression (p< 0.05), decreased the perfusion unit (p< 0.05), and patency rate (p< 0.001), compared with rats without thrombogenic anastomosis of femoral artery in both groups. In summary, this study demonstrates that microvascular anastomosis can be performed safely with reactive thrombocytosis alone without thrombogenic anastomosis. Meticulous microvascular anastomosis without triggering platelet activation is the most important factor to prevent thrombosed vessels in microsurgical anastomosis.