Study On The Construction And Performance Of Chitin-glucan Composite Hemostatic Sponge By In Situ Etching Mushroom Fruiting Body

Massive bleeding wound is the main cause of death in battlefield and life.Almost90%of the fatal injuries are non-compressible wounds and puncture wounds at the junction of limbs and trunk.Therefore,the preparation of hemostatic materials for these types of wounds is one of the current research challenges.In nature,mushroom fruiting bodies have a natural three-dimensional sponge structure without using any toxic crosslinking agents,and their main components are natural polymers such as glucan,chitin and protein that can be degraded in vivo and have excellent biosafety properties.In this study,the proteins and lipids on the surface of the Pleurotus eryngii were selectively removed by in situ etching through alkali treatment and chemical modification,while the chitin-glucan-based three-dimensional porous structure was retained to construct an efficient hemostatic sponge for non-compressible wounds.The effects of etching methods on the microstructure,water absorption,deformation response,mechanical properties and coagulation properties of hemostatic sponge were systematically investigated,and the structure-activity relationship was established.The biological safety of hemostatic sponge,hemostatic efficiency and wound healing in animal wound models were evaluated.The details are as follows:（1）Firstly,a chitin-glucan composite sponge（A-Sponge）with porous structure was produced by removing the hydrophobic protein layer and lipids from the outer surface layer of mushroom through alkali treatment,followed by surface modification of the composite sponge by TEMPO（2,2,6,6,-tetramethylpiperidin-1-yloxy radical）oxidation to introduce the hydrophilic group-COOH.Finally,ATC-Sponge was obtained by Ca²⁺crosslinking.ATC-Sponge had high water absorption（2400±300%）and excellent mechanical properties（σ=175 k Pa,ε=80%）.More importantly,ATC-Sponge had a blood-triggered shape memory function.In a rat liver injury model simulating bleeding from a puncture wound,ATC-Sponge showed an 89%reduction in bleeding volume compared to commercial gelatin sponges,demonstrating excellent hemostatic performance.In addition,the excellent wound healing ability of ATC-Sponge was confirmed by full-thickness wound healing experiment.In conclusion,a novel biodegradable hemostatic dressing was constructed for hemostasis of non-compressible wounds.（2）The above deproteinized A-Sponge was subjected to heterogeneous quaternization to obtain the quaternized modified sponge（AQ-Sponge）,which was subsequently cross-linked by tannic acid（TA）to obtain a hemostatic sponge（AQT-Sponge）with high water absorption（2047±105%）and excellent mechanical properties（σ=202 k Pa,ε=80%）.In the rat liver puncture injury model,AQT-Sponge effectively sealed the wound.At the same time,quaternary ammonium salt and TA in the sponge synergistically promoted the coagulation cascade,and the blood loss decreased from732±64 mg in the gelatin sponge group to 101±64 mg.In addition,the bactericidal performance of AQT-Sponge was higher than 95%for both S.aureus and E.coli due to the antibacterial activities of quaternary ammonium salts and TA.In summary,a highly effective hemostatic antibacterial sponge was prepared.