Ficin-Modified Chitosan Hydrogels: In Vitro Hemocompatibility and Hemostatic Performance

Abstract

Bleeding remains a leading cause of death in trauma and surgery, which drives the search for effective local hemostatic materials. Chitosan is a promising candidate because of its biocompatibility, biodegradability, and intrinsic hemostatic activity, yet chitosan hydrogels remain poorly characterized in terms of hemocompatibility. This study assessed the hemocompatibility and hemostatic activity of chitosan hydrogels with varying degrees of crosslinking and their complexes with ficin. Hydrogels were synthesized using 1,4-butanediol diglycidyl ether and characterized by ζ-potential measurements. Hemocompatibility was evaluated through hemolysis, hemoglobin spectroscopy, erythrocyte cytoarchitecture, and lymphocyte viability assays. Ficin was immobilized by adsorption and covalent binding, and hemostatic properties were assessed via swelling and kinetic blood coagulation. The hydrogels showed a statistically significant inverse dose-dependent hemolytic effect, attributed to their low colloidal stability. In contrast, hemoglobin spectra, erythrocyte cytoarchitecture, and lymphocyte viability showed no statistically significant changes. Adsorption immobilization provided higher protein loading and proteolytic activity than covalent binding. Ficin-containing compositions significantly accelerated blood coagulation compared with pristine hydrogels. These findings confirm the potential of the developed hydrogels and their ficin complexes as hemocompatible hemostatic materials.