Preparation method of bletilla striata polysaccharide-based electroactive composite sponge wound dressing

Abstract

The invention relates to the technical field of biomedical materials, in particular to a preparation method of a bletilla striata polysaccharide-based electroactive composite sponge wound dressing. The preparation method comprises the following steps: adding a proper amount of a CMCS solution into an rGO solution, and fully stirring; adding a CBM solution, and uniformly stirring; adjusting the pH value of the system to 6.9-7.4 by using triethanolamine; adding a Bsp solution into the system, and uniformly stirring; adding a proper amount of deionized water to adjust the system, and fully stirring; and pouring the obtained system into a mold, and freeze-drying to obtain the Bsp / GA-rGO composite sponge. The composite sponge dressing has excellent biological activity of bletilla striata polysaccharide and good compatibility of all components of an electroactive material, and the purpose of clinical conversion of the composite sponge dressing is achieved.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of biomedical materials, in particular to a preparation method of a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing. BACKGROUND

[0002] Wound healing is a complex and multifactorial regulated physiological process involving hemostasis, inflammatory response, cell proliferation, matrix remodeling and other stages, and wound dressing as a key auxiliary material for wound repair directly affects the healing efficiency and quality. Traditional dressings (such as gauze) can play a physical protective role, but have defects such as poor moisturizing property, easy adhesion to the wound surface, and inability to actively promote healing, and have been difficult to meet the treatment needs of complex wounds. Therefore, developing a new type of wound dressing with the functions of moisturizing, hemostasis, antibiosis and active repair promotion has become a research hotspot in the field of biomedical materials.

[0003] Natural polysaccharides are favored in the field of wound dressings due to their excellent biocompatibility, biodegradability and unique biological activity. Among them, Bletilla striata polysaccharide (Bsp) as a natural high molecular compound extracted from the tubers of traditional Chinese medicinal material Bletilla striata, has excellent properties such as biocompatibility, biodegradability, structural modifiability and self-assembly characteristics (Advances and prospects of Bletilla striata polysaccharide as promising multifunctional biomedical materials [J]. Materials & Design, 2022, 223: 111198.), and research shows that it also has good anti-inflammatory, antioxidant and wound healing properties (Exploration of the anti-inflammatory, analgesic, and wound healing activities of Bletilla Striata polysaccharide [J]. International Journal of Biological Macromolecules), and is an ideal wound dressing substrate. However, single Bletilla striata polysaccharide-based dressing has problems such as insufficient mechanical strength and limited antibacterial performance, which limits its clinical application.

[0004] In recent years, electroactive wound dressings have become a research frontier because they can simulate the endogenous electric field generated after human injury, promote cell migration, angiogenesis and collagen remodeling through electrical stimulation (Kloth LC. Electrical Stimulation Technologies for Wound Healing. Adv Wound Care (New Rochelle). 2014 Feb 1;3(2):81-90.). Commonly used electroactive materials include conductive polymers (such as polypyrrole and polyaniline), carbon-based materials (such as graphene and carbon nanotubes), and piezoelectric materials (such as polyvinylidene fluoride). However, most conductive materials have poor biocompatibility, poor compatibility with natural polysaccharide matrices, and easy aggregation, and the synergistic mechanism of their electroactivity and biological activity has not been clearly defined, making it difficult to maximize the healing effect of the composite dressing. SUMMARY

[0005] The present application aims to overcome the above-mentioned shortcomings of the prior art, such as the single function of Bsp-based dressings and the biocompatibility defects of electroactive materials, and provides a preparation method for a Bsp-based electroactive composite sponge wound dressing, which obtains a composite sponge dressing with excellent biological activity of Bsp and good compatibility of each component of the electroactive material.

[0006] The technical solution of the present application is as follows: A preparation method for a Bsp-based electroactive composite sponge wound dressing, characterized in that the preparation method is carried out according to the following steps: (1) Dissolve carbomer (CBM940) in deionized water, stir overnight, and obtain a carbomer solution for standby; dissolve carboxymethyl chitosan (CMCS) in deionized water, stir uniformly, and obtain a CMCS solution for standby; dissolve Bsp in deionized water, stir uniformly, and obtain a Bsp solution for standby; dissolve GA-rGO powder in deionized water, stir and ultrasonic to disperse uniformly, and obtain a rGO solution for standby; (2) Take an appropriate amount of CMCS solution and add it to the rGO solution, and stir thoroughly; (3) Mix the CMCS and rGO thoroughly, then add the mixed solution obtained in step (2) to the CBM solution, and stir uniformly; (4) Adjust the pH of the system to 6.9-7.4 with triethanolamine; (5) Add the Bsp solution to the system and stir uniformly at a speed of 600-800 r / min; (6) Add an appropriate amount of deionized water to adjust the system, and stir thoroughly; (7) Pour the obtained system into a mold, freeze-dry, and obtain a Bsp / GA-rGO composite sponge.

[0007] As preferred, the solution concentration of the Bsp solution in step (1) is 10%.

[0008] As preferred, the CMCS solution in step (1) is prepared by dissolving 0.1 g of carboxymethyl chitosan (CMCS) in 1 mL of deionized water, and stirring until uniform.

[0009] As preferred, the CBM solution in step (1) is prepared by dissolving 0.25 g of carbopol 940 (CBM940) in 20 mL of deionized water, and stirring until uniform.

[0010] As preferred, the preparation process of the GA-rGO powder in step (1) is as follows: at room temperature, 1.28 g of gallic acid (GA) is added to 80 mL of 0.2% GO solution, and after sufficient stirring, the mixed solution is placed in a reaction kettle and reacted at 95°C for 12 h. When the solution cools to room temperature, the GA-rGO powder is obtained by suction filtration and freeze-drying.

[0011] As preferred, the solution concentration of the rGO solution in step (1) is 0.4%-2%.

[0012] As preferred, the stirring temperature of steps (2) to (6) is 25°C.

[0013] As preferred, the stirring time of step (3) is 5 minutes.

[0014] As preferred, the stirring time of step (5) is 10-20 minutes to ensure that the system is fully cross-linked, and stirring is performed until a uniform and transparent hydrogel is obtained.

[0015] As preferred, the system adjustment in step (6) is performed by using deionized water at a ratio of 0.5 g of Bsp to 25 g of the total mass of the system.

[0016] As preferred, the freeze-drying time in step (7) is 36-48 h.

[0017] In steps (2)-(3), the CMCS needs to be mixed with rGO first, and then CBM is introduced. The order of addition allows rGO to be wrapped by anionic polysaccharides, avoiding direct contact with carbomer, which leads to agglomeration.

[0018] The beneficial effects of the present application are as follows: The present application overcomes the defects in the prior art such as insufficient mechanical strength of bletilla striata polysaccharide-based dressings, single function, poor compatibility of electroactive materials with natural polysaccharide matrix, limited synergistic healing effect, etc. Graphene (GA-rGO) powder reduced by gallic acid (GA) has good biocompatibility, electrical conductivity and antioxidant capacity; the Bsp / GA-rGO sponge has excellent mechanical properties, electrical conductivity, antioxidant properties, hemostatic properties and antibacterial properties; the synergistic effect of bioactivity and electrical stimulation is achieved, and the efficiency of wound repair is improved.

[0019] (1) Synergistic promotion of bioactivity and electrical stimulation: the present application organically combines the natural bioactivity of bletilla striata polysaccharide such as hemostasis and anti-inflammatory with the electrical signal regulation function of electroactive materials. Bletilla striata polysaccharide can quickly stop bleeding and inhibit the generation of inflammatory factors to reduce inflammatory reaction at the wound surface. Electroactive materials simulate endogenous electric field to promote directional migration of fibroblasts and vascular endothelial cells, accelerate granulation tissue formation and angiogenesis. The synergistic effect of the two can shorten the healing period and solve the problem of insufficient healing efficiency of single material.

[0020] (2) Optimized physical and mechanical properties: the Bsp / GA-rGO composite sponge has a porous 3D structure, which meets the characteristics of high absorption materials, and the sponge has good adhesion after absorbing water. Through compression-tension experiments, it is found that the composite sponge has excellent mechanical properties.

[0021] (3) Good biological safety and antibacterial properties: the material is verified by in vitro experiments to have no cytotoxicity and can inhibit the growth of escherichia coli and staphylococcus aureus.

[0022] (4) Efficient preparation process and easy industrialization: the "solution blending-freeze drying" process used in the present application does not require complex equipment, has mild reaction conditions, is suitable for large-scale production, and reduces the cost of clinical application. BRIEF DESCRIPTION OF DRAWINGS

[0023] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or prior art description will be briefly introduced below. Obviously, other drawings can also be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0024] Figure 1 SEM image of the bletilla striata polysaccharide / gallic acid-reduced graphene sponge of the present application.

[0025] Figure 2 SEM image of the composite sponge of Comparative Example 1 which is not added according to the defined order.

[0026] Figure 3 SEM image of the bletilla striata polysaccharide sponge of the present application.

[0027] Figure 4 Chart of the conductivity of the white psuedostellaria heterophylla polysaccharide-based electroactive composite sponge wound dressing with different reduced graphene contents in the present application.

[0028] Figure 5 Chart of the tensile properties of the white psuedostellaria heterophylla polysaccharide-based electroactive composite sponge wound dressing with different reduced graphene contents in the present application.

[0029] Figure 6 Chart of the compressive properties of the white psuedostellaria heterophylla polysaccharide-based electroactive composite sponge wound dressing with different reduced graphene contents in the present application.

[0030] Figure 7 Cell toxicity imaging chart of the white psuedostellaria heterophylla polysaccharide / gallic acid-reduced graphene sponge 24 h and 72 h in the present application.

[0031] Figure 8 Chart of the antibacterial ability experiment of the white psuedostellaria heterophylla polysaccharide-based electroactive composite sponge wound dressing with different reduced graphene contents in the present application.

[0032] Figure 9 Chart of the hemostatic ability of the white psuedostellaria heterophylla polysaccharide / gallic acid-reduced graphene sponge in the present application. DETAILED DESCRIPTION

[0033] In order to enable personnel in the technical field to better understand the technical solutions in the present application, the technical solutions in the embodiments of the present application will be described clearly and completely in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor should be within the scope of protection of the present application.

[0034] Embodiment 1 A preparation method of a white psuedostellaria heterophylla polysaccharide-based electroactive composite sponge wound dressing, characterized in that the preparation method is performed according to the following steps: (1) 0.125g Carbomer 940 was dissolved in 10ml water, stirred overnight at room temperature, to obtain Carbomer solution for standby; 0.1g carboxymethyl chitosan was dissolved in 1ml deionized water, stirred uniformly, to obtain CMCS solution for standby; 0.5g Bletilla striata polysaccharide was dissolved in 5ml deionized water, stirred uniformly, to obtain Bsp solution for standby; CBM solution was prepared by dissolving 0.25g Carbomer 940 (CBM940) in 20ml deionized water, stirring uniformly; 1.28g gallic acid (GA) was added to 80ml 0.2% GO solution at room temperature, and after being stirred thoroughly, the mixture was put into a reaction kettle and reacted at 95℃ for 12h; when the solution was cooled to room temperature, it was filtered, freeze-dried to obtain GA-rGO powder, 0.08g GA-rGO powder was dissolved in 5ml deionized water, stirred and ultrasonicated to disperse uniformly, to obtain rGO solution for standby; (2) 0.625ml 10% carboxymethyl chitosan solution was added to 5ml rGO solution and stirred thoroughly; (3) CMCS was mixed with rGO thoroughly, then CBM was introduced, the solution of step (2) was added to CBM solution, and stirred for 5min to make it uniform; (4) The pH of the system of step (3) was adjusted to 6.9-7.4 with triethanolamine; (5) 10% Bsp solution of step (1) was added to the system, and stirred at a speed of 600-800r / min for 15min to make it uniform; (6) The system was adjusted to 25g with deionized water, and stirred at room temperature for 10min to make it thoroughly stirred; (7) The obtained sample was poured into a mold, and freeze-dried for 48h to obtain Bsp / GA-rGO composite sponge.

[0035] Comparative Example 1 The difference from Example 1 is that step (2) is changed to adding rGO solution of step (1) to CBM solution and stirring thoroughly; step (3) is changed to adding 0.625ml 10% CMCS solution to the system of step (2). Figure 2 As shown in the SEM picture of the composite sponge, the graphene occurs in lamellar stacking and does not form a porous structure.

[0036] Comparative Example 2 Preparation of Bletilla striata polysaccharide sponge wound dressing The difference from Example 1 is that there is no addition of rGO solution in step (4) and step (5) of Example 1. Figure 3As shown, SEM observation revealed that the surface of the Bletilla striata polysaccharide sponge was relatively smooth, with a small number of wrinkles in some areas, uneven pore distribution, and pore wall collapse in some areas. It was not conductive, and its mechanical properties, antibacterial ability, and hemostatic ability were all inferior to those of the rGO-containing composite sponge.

[0037] Comparative Examples 3-5 Preparation of Bletilla striata polysaccharide-based electroactive composite sponge wound dressing The difference from Example 1 is that in step (4) of Example 1, the amounts of GA-rGO added were 0.02g, 0.06g, and 0.1g, respectively. Figure 4 , 5 As shown in Figures 6 and 7, its electrical conductivity and mechanical properties are not as good as those of GA-rGO when the mass is 0.08g.

[0038] Experimental Analysis: 1. Microstructure of Bletilla striata polysaccharide-based electroactive composite sponge observed by scanning electron microscopy (SEM) The composite sponge prepared in Example 1 was cut into small pieces of 1cm × 1cm × 0.2cm and fixed to the SEM sample stage with conductive adhesive. It was then subjected to gold sputtering (sputtering time 60s, gold layer thickness approximately 5nm) to ensure good conductivity. The sample was observed using a scanning electron microscope with an accelerating voltage of 5kV and a working distance of 8-10mm. The surface and cross-sectional morphology of the sample were observed at low magnification (50×, 200×) and high magnification (1000×, 5000×). Figure 1 As shown, the composite sponge surface exhibits a fluffy and porous 3D structure, indicating that the composite sponge prepared by the freeze-drying process in this application has high porosity, a three-dimensional interconnected structure, and uniform dispersion of electroactive materials. This structure is beneficial for the absorption of wound exudate, cell infiltration, and nutrient exchange, verifying the effective control of the microstructure of the material by the preparation method.

[0039] 2. Detection of the conductivity of Bletilla striata polysaccharide-based electroactive composite sponge The composite sponge prepared in Example 1 was cut into cuboids of 1cm × 2cm × 0.5cm and dried in a vacuum drying oven at 60℃ for 24 hours until constant weight was achieved to eliminate the influence of moisture on conductivity. The sample was tested using a dual-probe method on an electrochemical workstation (CHI760E), and the conductivity (κ, S / cm) was calculated according to the following formula.

[0040] κ=(I / V)(L / A) Where V is the measured voltage, and I is the current provided by the potential state, in A (cm). 2) is the cross-sectional area of the sample, and L (cm) is the distance between the two probes. The conductivity of the composite sponge in Example 1 was measured to be 0.23 S / cm, with three repeated measurements and averaging, as detailed in Figure 4 .

[0041] The conductivities of the composite sponges with rGO solution concentrations of 0.4%, 1%, and 2% were 0.06 S / cm, 0.16 S / cm, and 0.15 S / cm, respectively, as detailed in Figure 4 .

[0042] 3. Mechanical properties of the Bletilla striata polysaccharide-based electroactive composite sponge A texture analyzer (TA.XT Plus) was used to test the tensile, compression, and compression-recovery properties of the composite sponge.

[0043] (1) Tensile properties: The composite sponge in Example 1 was cut into a strip-shaped sample with dimensions of 5 cm in length, 1 cm in width, and 0.5 cm in thickness. An A / TG tensile probe (50 N sensor) was selected, and the test mode was single-axis tensile test. The parameters were as follows: pre-test speed: 5 mm / s, test speed: 3 mm / s, post-test speed: 10 mm / s, trigger force: 2 g, tensile distance set to "stop automatically when the sample breaks", data acquisition rate: 100 pps. The strip-shaped sponge was fixed at the upper and lower clamps of the texture analyzer, ensuring that the sample had no wrinkles and no pre-tension, and the clamp distance was 30 mm. The instrument was started, and the upper clamp moved upward at the preset speed to stretch the sample axially until it broke. The test results showed that the tensile strain range of the composite sponge could reach about 80%, as detailed in Figure 5 .

[0044] (2) Compression properties: The composite sponge in Example 1 was cut into a cylindrical sample with a diameter of 1.5 cm and a height of 1 cm. The cylindrical sponge was placed horizontally on the center of the texture analyzer stage, and the probe position was adjusted to be coaxial with the sample to avoid eccentric compression. The instrument was started, and the probe descended at the pre-test speed, contacted the sample, and triggered a 10 g force. Then, the sample was compressed at a speed of 1 mm / s to 80% deformation to reach the preset compression amount. Immediately after, the probe rose to the initial position at a speed of 2 mm / s to complete a single compression-recovery process. The test results showed that the compression stress in the strain range of 60%–80% could reach more than 120 kpa, as detailed in Figure 6 .

[0045] (3) Compression-cycling performance: Take the composite sponge in Example 1, make the sample into a cylinder with a diameter of 1.5 cm and a height of 1 cm, place the sponge sample horizontally on the material tester loading platform, adjust the probe position so that the center of the probe is aligned with the center of the sample, start the instrument, and the probe will descend at the preset speed, contact the sample surface and trigger 5g force to start compression until 80% deformation is reached; keep the compression state for 1s, then the probe will rise to the initial position at the test speed, complete the first compression; after 5s, repeat the above compression process, complete the second cycle, a total of four cycles. Test results show that after four cycles, the composite sponge can still basically recover to the initial shape.

[0046] 4. Detection of cell toxicity of bletilla striata polysaccharide-based electroactive composite sponge (1) CCK8 experiment: Mouse fibroblasts (L929 cells) were cultured in a 5% CO2, 37°C constant temperature incubator for 24h, the composite sponge was broken by a wall breaking machine and then irradiated for sterilization, dispersed into complete DMEM medium, and then configured into a medium suspension with concentrations of 1 / 5 / 10 mg / mL, and a pure complete DMEM medium group was set as a control. Before treatment, the cells were inoculated into a cell culture 96-well plate and cultured for 16h. Take L929 cells in the logarithmic growth phase, count the cells, and adjust the cell concentration to 5 × 10 3 cells / well. All treatments were performed at 37°C and exponential growth density. According to the above grouping, remove the DMEM complete culture medium and replace it with 100 μL of the prepared sample working solution, and incubate in a 5% CO2, 37°C constant temperature incubator for 24h. Remove the culture medium, wash each well with 100 μL of PBS twice, with a 3min stay each time. Remove the PBS and add 100 μL of culture medium containing 10% CCK8 (10 μL), and incubate in a 5% CO2, 37°C constant temperature incubator for 1h. Take out the culture plate, take a photo, and use an enzyme marker to collect absorbance values at 450nm wavelength, with 3 replicates per group. The results show that the CCK-8 method detects that the bletilla striata polysaccharide-based electroactive composite sponge has no obvious toxicity to L929 cells (toxicity grade 0).

[0047] (2) Dead and live cell staining laser confocal experiment: Before treatment, the cells were inoculated into a confocal dish and cultured for 16h. Take L929 cells in the logarithmic growth phase, count the cells, and adjust the cell concentration to 2×10 6per well. All treatments were performed at 37 °C and at an exponential growth density. The DMEM complete medium was removed and replaced with 1 mL of the prepared sample working solution, and incubated in a 37 °C constant temperature incubator with 5% CO2for 24, 72, and 120 h. The medium was aspirated, and the cells were gently washed 3 times with preheated PBS at 37 °C. PBS containing 0.1% Calcein-AM and 0.3% PI was added, and the cells were incubated at 37 °C for 20 min in the dark. The staining solution was aspirated, and the cells were washed 3 times with PBS. Imaging was performed immediately. Imaging was collected under a laser confocal microscope at 20 / 40 magnification and at excitation wavelengths of 490 nm and 545 nm. A large number of green fluorescent living cells were observed at 24 h, 72 h, and 120 h, and the cells were spindle-shaped or polygonal in shape, with clear nuclei and no obvious red fluorescence. The results were consistent with the CCK8 experiment, indicating that the BTP-based electroactive composite sponge had no obvious cytotoxicity. See Figure 7 .

[0048] 5. Detection of the antibacterial ability of the BTP-based electroactive composite sponge 2 mL of bacterial solution with a concentration of 10 4 CFU / mL was mixed with 30 mg of the composite sponge for co-culture. The bacterial solution was cultured in a shaking bed at a temperature of 37 °C and a rotation speed of 200 rpm for 2 h, and then diluted 1000-fold. 100 μL was taken and spread on solid agar medium. After completion of the culture, the bacterial solution was diluted 10 times successively with sterile PBS solution, and 100 μL of the diluted solution was uniformly spread on LB solid medium. The medium was placed in a 37 °C constant temperature incubator and cultured for 18 h. The medium was taken out, photographed, and the number of bacteria was recorded.

[0049] The results of the plate spread test control group showed that the composite sponge had a certain antibacterial effect on Staphylococcus aureus and Escherichia coli. See Figure 8 .

[0050] 6. Hemostatic ability of the BTP-based electroactive composite sponge After the rats were anesthetized, they were fixed, the hair on their abdomens was shaved, the liver was exposed, and the tissue fluid around the liver was wiped with filter paper. The weighed filter paper was placed under the liver, and a 1 cm long and 2 mm deep wound was made on the liver with a surgical knife. Then, a 1.5 cm * 1.5 cm sponge was used to cover the wound. The filter paper was weighed at 1 min, 2 min, and 3 min, until the bleeding stopped completely. Finally, the amount of blood loss in the rats was calculated. The results showed that the amount of blood loss in the control group without treatment was about 500 mg, and the amount of blood loss in the composite sponge group was only about 60 mg, indicating that the composite sponge had very excellent hemostatic ability. See Figure 9 .

Claims

1. A method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing, characterized in that, The preparation method is carried out according to the following steps: (1) Dissolve carbomer (CBM940) in deionized water and stir overnight to obtain a carbomer solution for later use; dissolve carboxymethyl chitosan (CMCS) in deionized water and stir evenly to obtain a CMCS solution for later use; dissolve Bsp in deionized water and stir evenly to obtain a Bsp solution for later use; dissolve GA-rGO powder in deionized water, stir and sonicate to disperse it evenly to obtain an rGO solution for later use; (2) Add an appropriate amount of CMCS solution to the rGO solution and stir thoroughly; (3) After CMCS and rGO are thoroughly mixed, the mixed solution obtained in step (2) is added to the CBM solution and stirred until homogeneous; (4) Adjust the pH of the system to 6.9-7.4 using triethanolamine; (5) Add the Bsp solution to the system and stir evenly at a speed of 600-800 r / min; (6) Add an appropriate amount of deionized water to adjust the system and stir thoroughly; (7) Pour the obtained system into a mold and freeze-dry it to obtain Bsp / GA-rGO composite sponge.

2. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: In step (1), the concentration of the Bsp solution is 10%.

3. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: In step (1), the CMCS solution is prepared by stirring the solution evenly at a ratio of 0.1g of carboxymethyl chitosan (CMCS) to 1mL of deionized water.

4. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: In step (1), the CBM solution is prepared by stirring the solution evenly at a ratio of 0.25g of carbomer 940 (CBM940) to 20mL of deionized water.

5. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The preparation process of GA-rGO powder in step (1) is as follows: at room temperature, 1.28g gallic acid (GA) is added to 80mL of 0.2% GO solution, and the mixture is stirred thoroughly to obtain a mixed solution. The solution is then placed in a reaction vessel and reacted at 95℃ for 12h. After the solution is cooled to room temperature, it is filtered and freeze-dried to obtain GA-rGO powder.

6. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The concentration of the rGO solution in step (1) is 0.4%-2%.

7. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The stirring temperature in steps (2) to (6) is 25°C.

8. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The stirring time in step (3) is 5 minutes.

9. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The stirring time in step (5) is 10-20 minutes to ensure that the system is fully cross-linked and stirred until it is a uniform and transparent hydrogel.

10. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The adjustment of the system in step (6) is to use deionized water at a ratio of 0.5g Bsp to 25g of total system mass.

11. The method for preparing a Bletilla striata polysaccharide-based electroactive composite sponge wound dressing according to claim 1, characterized in that: The freeze-drying time in step (7) is 36 h - 48 h.

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