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Free flow hemostatic agents are dominating over non-flowable hemostats due to their ability to cover asymmetrical wound surfaces of any depth and easily remove excess materials with irrigation. The objective of this study was to evaluate the activation of a coagulation system both in vitro and in vivo. We assessed detailed physical characteristics of a microbial transglutaminase (m-TG) crosslinked thrombin (T laden Gelatin (Gel) hemostat sealant in vitro and its hemostatic efficacy for controlling bleeding caused by liver trauma in rat