BACKGROUND: A high activation level is a normal characteristic of lyophilized platelets (LPs); however, the effects of this activation level on the efficacy and safety of LPs after transfusion are debated. OBJECTIVES: We aimed to test the efficacy and safety of pre-activated LPs (PLPs) in rabbits with traumatic bleeding and shock. METHODS: In vitro characteristics of PLPs, including activation level, aggregation, migration, and thromboelastography parameters, were evaluated. Limb soft tissue injury accompanied by seawater immersion and controlled hemorrhagic shock was induced in 50 rabbits, which were then divided into five groups: A (no resuscitation), B (resuscitation with Lactated Ringer's solution, LR), C (resuscitation with LR and fresh platelets), D (resuscitation with LR and LPs), and E (resuscitation with LR and PLPs pre-activated by thrombin). Blood loss, platelet count, blood urea nitrogen and lactic acid concentrations, and in vivo thromboelastography value and maximum amplitude were recorded. Biotin-X-N-hydroxysuccinimide labeling and flow cytometry were used to measure the number of infused platelets left in circulation. Histology was used to assess whether aberrant thrombi were formed in the kidney, lung, or liver. RESULTS: PLPs exhibited an increased P-selectin level, enhanced aggregation, and shortened values, with no obvious changes in migration ability or maximum amplitude. PLPs transfusion had a non-inferior effect on all in vivo parameters compared with fresh platelet transfusion, and the circulation time of PLPs was much shorter than that of fresh platelets. No obvious thrombi were found. CONCLUSIONS: PLPs transfusion demonstrated non-inferior efficacy and safety compared with fresh platelet transfusion.