Bletilla striata polysaccharide (BSP) and chitosan (CS) were chemically cross-linked using oxalyl chloride to prepare a composite hemostatic sponge (BSP-CS), and the process parameters were optimized using the Box–Behnken design (BBD) with response surface methodology. To optimize the performance of the hemostatic sponge, we adjusted the ratio of independent variables, the amount of oxalyl chloride added, and the freeze-dried volume. A series of evaluations were conducted on the hemostatic applicability of BSP-CS. The characterization results revealed that BSP-CS had a stable bacteriostatic effect on Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa within 72 h, and the bacteriostatic rate was above 30%. The CCK-8 cytotoxicity test demonstrated that BSP–CS had a certain effect on promoting cell proliferation of L929 cells. In the mouse tail-cutting experiment, the hemostasis time of BSP–CS was 463.0 ± 38.16 s, shortened by 91.3 s on average compared with 554.3 ± 34.67 s of the gauze group. The blood loss of the BSP–CS group was 28.47 ± 3.74 mg, which was 34.7% lower than that of the control gauze group ( 43.6 ± 3.83 mg). In the in vitro coagulation experiment, the in vitro coagulation index of the BSP–CS group was 97.29 % ± 1.8 %, which was reduced to 8.6% of the control group. The CT value of the BSP–CS group was 240 ± 15 s, which was 155 s lower than that of the gauze group ( 355 ± 31.22 s). All characterization results indicate that BSP–CS is an excellent hemostatic material.

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See supplementary material online for the details of the batch preparation process of BSP, the content of polysaccharide in BSP powder and the molecular weight of BSP.

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