Functional effect of hemostatic pad based on a collagen with chitosan by addition of batroxobin

Abstract

Uncontrolled excessive bleeding in daily life, whether due to sudden accident, surgical operations, or combat may cause infectious complications that may lead to death by shock. Therefore, topical hemostats and sealants are required to assist effective treatment, arrest bleeding, and stabilize the casualty before definitive care.

This study use collagen and chitosan, which are both natural materials and are one of the most widely used hemostats these days. The materials are role in platelet activation, adhesion and aggregation. The use of chitosan helps the blood coagulation as it combines with red blood cells. However, these materials have been reported to lack sufficient hemostatic effect in a timely manner.

Hence, in this research, a novel pad made a faster and more effective sponge type hemostats by using the recombinant batroxobin, a venomous component from the snake Bothrops atrox moojeni, which transforms the fibrinogen to fibrin in the process of hemostasis; it expected that recombinant batroxobin would support natural substances to achieve a synergetic effect of hemostasis performances. First, collagen and chitosan were dissolved with batroxobin in acetic acid for fabrication of a novel hemostatic pad and then characteristic analysis using Scanning Electron Microscope. Fabricated sponge groups were collagen-only, chitosan-only, collagen with chitosan, collagen or chitosan containing recombinant batroxobin 1, 2 and 3 BU/well for in vitro studies, collagen or chitosan containing recombinant batroxobin 3 and 5 BU/well for in vivo studies and collagen with chitosan containing recombinant batroxobin 1, 2, 3 and 5 BU/well. Fibrinogen assay and platelet activation assays represented respective hemostats mechanism, even combination of the materials. Whole blood clotting assay and animal experiments investigated blood coagulation activities in the injury sites. In addition, H&E and PTAH staining methods for histological analysis. Through the experimentations result, a novel hemostatic pad confirmed that the substances maintained to their hemostatic properties in the pad, also coagulation studies proved pad effectively reduced blood loss and shortened bleeding time by addition of batroxobin.

In conclusion, the novel hemostatic pad is reasonable that an enormous potential for excessive bleeding injury and improve effects of natural substance hemostatic pad when it is cooperated with recombinant batroxobin.