Evaluation of hemostatic effects of an adhesive with recombinant batroxobin

Abstract

Uncontrolled bleeding can cause serious complications in surgical procedures, traumatic injuries on battle fields and in daily life. Excessive blood loss could ultimately threaten human life. Therefore, studies concerning hemostatic materials and their therapeutic solutions have been undertaken to arrest bleeding within a relevant time to minimize blood loss. This research focused on a topical hemostatic adhesive which is composed of powdery, biodegradable adhesive and recombinant batroxobin. Batroxobin is derived from the venom of a snake, Bothrops atrox moojeni, and has an active property for hemostasis. In the blood coagulation system, batroxobin converts fibrinogen to fibrin, which was represented by a fibrinogen clotting assay with the batroxobin-containing adhesive. Also, whole blood obtained from an SD rat was examined to determine whether it could form clots on the hemostatic adhesive. Through in vivo experiments, activities of the adhesive on the bleeding sites of the animals were evaluated with thirty-two SD rats and thirty-two ICR mice. The animals were placed into four groups (n=8). One group was left untreated and the other groups were allocated adhesives containing 0, 30 or 50 NIH U/ml batroxobin respectively. To compare the amount of blood loss, the liver wound model was applied. The result was histologically analyzed by H&E and PTAH staining methods. The femoral artery injury model of rats was applied for analysis of the hemostasis times and patterns. The results of the liver model indicated that blood loss was reduced with the treatment of those mice that were allocated adhesive containing increasing concentrations of batroxobin. The time required for hemostasis was also significantly decreased with application of the adhesive compared to the control. The 50 NIH U/ml batroxobin-containing adhesive was remarkably effective for reduction of coagulation time. Also, the addition of batroxobin could decrease the occurrence of re-bleeding during the hemostatic procedure with the adhesive. The biodegradable adhesive played a role as the first mechanical barrier against bleeding; while batroxobin was directly involved in formation of blood clots to arrest bleeding. Hence, the adhesive promoted hemostasis successfully in both types of bleeding injuries with an increase in the concentration of batroxobin. In conclusion, an adhesive containing the appropriate concentration of batroxobin would be effective in controlling bleeding with respect to the amount of blood loss and the blood arresting time.