N-succinyl chitosan-oxidized hyaluronic acid-calcium chloride hydrogel as hemostatic agent

This study aims to develop an effective hemostatic agent in the management of irregular and deep wounds that can accelerate the hemostatic process. The background revealed the importance of rapid treatment of bleeding, with data showing a significant risk of death from blood loss. Current treatments use conventional hemostatic dressings, but they are less effective on irregular surgical wounds. Several studies have developed chitosan, hyaluronic acid, and CaCl₂-based hydrogels that have hemostatic, regenerative, and antibacterial potential. However, there is still a need to develop hydrogels that are thermally stable, biocompatible, and able to accelerate the hemostatic process. This research will synthesize self-healing hydrogels by modifying the structure of chitosan and hyaluronic acid, using a certain ratio of ingredients. The research procedure was carried out with the preparation of N-succinyl chitosan (NSC) and oxidized hyaluronic acid (OHA) as the main ingredients which were then added with CaCl₂ to produce self-healing injectable hydrogel. First, NSC and OHA were dissolved in phosphate buffer solution (pH = 7.4 PBS) to obtain 60 mg/mL NSC and OHA solution respectively. Calcium chloride was then dissolved in water to obtain 120 mg/mL CaCl₂ solution. Then NSC-OHA-CaCl₂-based hydrogels were synthesized through rapid and full solution mixing above room temperature with the composition of (1-1-0.1; 1-1-0.2; and 1-1-0.3). The targeted findings of this research are sample characterization results that explain and prove the best NSC-OHA-CaCl₂ composition variation that can be used as a hemostatic agent for irregular and deep wounds. The results of the analysis obtained FTIR test data with the formation of C = N functional groups in the four samples; blood clotting time test for sample K0, K1, K2, and K3 with time 4.6, 3.33, 2.66, and 1 s; MTT assay with cell viability percentage of 77.82% for sample K0, 84.18% for sample K1, 89.30% for sample K2, and 89.50% for sample K3; hemolysis index percentage of 0.373% for sample K0, 0.555% for sample K1, 0.625% for sample K2, and 0.201% for sample K3; Viscosity test obtained data of 13 dPa s for sample K0, 15 dPa s for sample K1, 16 dPa s for sample K2, and 18 dPa. The injectability test yielded an injectability percentage of 96.84% for sample K0, 95.03% for sample K1, 94.78% dPa s for sample K2, and 94.61% for sample K3; the DSC test results of the four samples obtained a transition peak at the exothermic peak of 62.27°C for sample K0, 70.23°C for sample K1, 73.77°C for sample K2, and 74.49°C for sample K3; and the characteristic graph of the TGA test results, the weight profile of the hydrogel during heating which showed a mass change of 21.64 mg in sample K0, 16.89 mg in sample K1, 15.37 mg in sample K2, and 11.43 mg in sample K3 (°C).