Hemostatic properties of a venomic protein in rodent dermal injuries

Hemostatic properties of a factor Xa-like protease (Q8009) from the Australian snake Pseudonaja textilis textilis were determined. In tail-tip transection and dermal incision (hind limb) models, reagents were applied with collagen matrix. Blood was collected on filter paper chads for 12 one-minute intervals or until hemostasis. Determination of blood loss was performed using the hematin content and reported as blood loss per minute and total blood lost. Results from the studies demonstrated that the addition of the protease Q8009 and collagen matrix significantly reduced the volume of blood loss and shortened the time-to-hemostasis. In the dermal incision model, Q8009 (100, 250 and 1000 microg/ml) plus collagen matrix significantly reduced (p<0.001) the volume of blood lost relative to Thrombin and shortened the time-to-hemostasis to 2.0 min compared to 4.77 min with Thrombin. In the tail-tip transection model when Q8009 was mixed with a collagen matrix there was no significant reduction in blood loss, when compared to Thrombin plus collagen matrix. However, when injured tail-tips were held in Q8009 (1000 microg/ml) solution, there was a significant reduction (p<0.001) in blood loss (5.88 microl) versus that of Thrombin at 58.0 mul, and time-to-hemostasis was reduced from 11 min with Thrombin to 3 min when the Q8009 solution was used. In these studies, topical application of the venomic protease Q8009 significantly reduced total blood loss with a shorter time-to-hemostasis relative to Thrombin.