Fabrication and characterization of hemostatic chitosan/gelatin cryogel containing verbascum thapsus extract and investigation of hemostatic effect

Hemorrhage is one of the biggest threats to human life and it causes approximately 40% of trauma deaths. To control bleeding, hemostatic dressings combining different polymers have gained interest in recent years. In this study, chitosan/gelatin hemostatic cryogels containing Verbascum Thapsus was fabricated, characterized morphologically, chemically, and biologically, and investigated hemostatic activity in vitro. Cryogels having unique porosity, rapid absorption, and cell infiltration characteristics are suitable for hemostatic applications. The combination of chitosan and gelatin creates a matrix enhancing platelet adhesion and aggregation via functional groups, whereas the use of V. thapsus extract increases bioactivity to accelerate clot formation and shorten coagulation time. The interconnected macroporous structure and average pore diameters of 225-478 µm were observed by SEM. The cryogels showed a high swelling ratio of 3350%. The V. Thapsus containing cryogels demonstrated bacterial inhibitions up to 89% against E. coli and 78% against S. aureus. The cell viability of the cryogels was investigated by a human fibroblast cell line. The blood compatibility of cryogels was proved with the 1% hemolysis ratio. The blood hemostatic activity of V. thapsus extract was investigated by using an in vitro whole blood clotting assay. The blood clotting index (BCI) of the chitosan/gelatin cryogels was improved from 11.9 to 6.5 with adding V. thapsus extract and the clotting time was decreased. The V. thapsus extract containing chitosan/gelatin cryogels demonstrated great hemostatic potential for hemorrhage applications.