Optimization Preparation and Evaluation of Chitosan Grafted Norfloxacin as a Hemostatic Sponge

Considering the great harm to the human body caused by severe and massive bleeding, in this study, chitosan-grafted norfloxacin (CTS-NF) composites were prepared with chitosan (CTS) and norfloxacin (NF) as raw materials by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-mediated coupling method to solve the limitations of slow hemostatic and poor anti-infective effects of current dressings on the market. The effects of the mass ratio of CTS to NF (M(CTS)/M(NF)), reaction temperature T and reaction time t on the grafting rate (η%) of the products were investigated through single factor tests. The preparation process was optimized with the η% as an evaluation index by means of the Box–Behnken test design and response surface analysis. The antimicrobial activity was evaluated by inhibition zone assay, and the hemostatic activity of the prepared composites was evaluated in vitro and in vivo. The results suggested that the optimum preparation conditions were the mass ratio of CTS to NF (M(CTS)/M(NF)) 5:3, reaction temperature 65 °C, and reaction time 4 h. Under this condition, the η% of CTS-NF was 45.5%. The CTS-NF composites displayed significant antimicrobial activities. Moreover, in vitro hemostasis results revealed that the CTS-NF composite had a lower blood clotting index and absorbed red blood cells to promote aggregation. In vivo ear and live hemostasis, the CTS-NF groups showed short hemostatic time (49.75 ± 3.32 s and 50.00 ± 7.21 s) and more blood loss (0.07 ± 0.010 g and 0.075 ± 0.013 g). The results showed that CTS-NF reduced the bleeding time and volume, exhibiting a significant coagulation effect. Therefore, the CTS-NF sponge is expected to be a new, effective hemostatic and antibacterial material in the future.