| Uncontrolled bleeding is an important cause of military and civilian casualties.Therefore,rapid and effective hemostasis can not only reduce the suffering of the injured,but also reduce the risk of injury.As a new potential hemostatic material,Graphene oxide(GO)has received more attention in the field of hemostasis,because of its good physical and chemical properties.However,due to its potential for thrombosis,hemolysis and cytotoxicity,pure GO is subject to various limitations in the application of hemostatic materials.Bletilla striata belongs to the Orchidaceae family,and has been widely used as an astringent hemostatic medicine in traditional Chinese medicine(TCM)for thousands of years,TCM holds that it can treat hematemesis,hemoptysis,traumatic hemorrhage and ulcerative carbuncles.Bletilla striata polysaccharide(Bsp),traditionally used constituents of Bletilla striata,exhibited good biocompatibility,biomedical activity,low toxicity,absorbability and degradability.Such polysaccharide can be structurally modified dete to the presence of abundant hydroxyl and long chains.Therefore,Bsp is an excellent candidate for GO structure modification.Therefore,we present a simple,rapid,environmentally and low-cost method to develop a new type of hemostatic material Bletilla striata polysaccharide modified graphene oxide composite sponge(BGCS),and its physicochemical properties were characterized.The biocompatibility and cytotoxicity were investigated,and the Rat-tail amputation and whole blood clotting evaluation were conducted.This study provided a reference for the development of Bletilla striata polysaccharide modified graphene oxide materials.The experimental contents include the following:1.The graphene oxide was synthesized by the modified Hummers method,the excess acid was removed by dialysis treatment for 7 days,and the viscous liquid was ultrasonicated for 2 h to obtain a uniform GO dispersion2.The purchased Chinese herbal medicine Bletilla striata were dried,pulverized,degreased,extracted with water and alcohol,combined with Sevag deproteinization technology to extract the Bletilla striata polysaccharide,and the precipitate was washed with absolute ethanol,acetone and diethyl ether to obtain refined Bletilla striata polysaccharide.3.Bletilla striata polysaccharide(16 mg/mL)and GO solution were mixed uniformly by simple solution blending,and lyophilized to prepare Bsp modified GO composite sponge.By changing the mass ratio of Bsp to GO,from 1:1,1:5,1:10 to 1:15,accordingly BGCS,BGCS-5,BGCS-10 and BGCS-15 were prepared.4.Physicochemical properties were characterized by scanning electron microscopy(SEM),infrared spectrometer(IR),Raman spectroscopy(RAMAN),X-ray diffraction(XRD),Zeta potential analyzer,thermogravimetric analyzer(TG),porosity and water absorption.5.Its safety and biocompatibility were evaluated by in vitro hemolysis test and cytotoxicity test.6.The hemostatic properties of the materials were evaluated by whole blood clotting evaluation,rat-tail amputation,blood cell morphology studies,and blood cell and platelet selective adhesion experiments.The results showed that GO and Bsp cross-linked in the BGCS to form a uniform,fluffy porous cavity structure,showing good biocompatibility(hemolytic rate < 2%,low cytotoxicity).In addition,the BGCS's porosity more than 90% and showed good water absorption capacity(5239.86 ± 129.65 %).In the absence of anticoagulant,BGCS can promote coagulation within 30 s,with good hemostasis.Further studies of coagulation mechanisms have shown that BGCS has a high charge(-27.3 ± 0.9 mV)and exhibits strong platelet stimulation.In addition,BGCS can induce the blood cell aggregation,accelerate fibrin formation and accelerate blood coagulation.In the synergistic effect of the hemostasis mechanism described above,the BGCS can stop bleeding in rat-tail amputation within 50 s.Therefore,the BGCS provides a new perspective for the safe application of GO in the field of trauma hemostasis. |
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