Oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel, and preparation method and application thereof

Abstract

The application discloses an oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel, which comprises oxidized konjac gum, hydrazide hyaluronic acid, neomycin sulfate, cannabidiol and 2-cresyl alcohol. The application further provides a preparation method of the oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel and application of the oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel in preparation of hemostatic materials. The composite hydrogel has high adhesion and excellent antibacterial property, is suitable for various wound surfaces, and has excellent hemostatic, analgesic, infection-reducing and wound healing-promoting effects.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomedical materials, and particularly relates to an oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel as well as a preparation method and application thereof. BACKGROUND

[0002] Hemostasis and healing of tissue wounds, especially for massive hemorrhage, and the demand for related hemostatic materials are still problems to be solved at present. Various complications such as infection and inflammation often occur during wound healing, which particularly increases the biological and pharmacological requirements for hemostatic materials. The commonly used gauze and wound dressing is easy to fall off during wound healing, and causes secondary injury due to adhesion to the wound after absorbing body fluid, and does not have the high efficiency of hemostasis and wound healing promotion.

[0003] The hydrogel dressing as a kind of hydrophilic biomaterial can act as a barrier for the skin of the wound, can stop bleeding, and can also load various effector molecules to effectively prevent wound infection, but the current hydrogel dressing has various problems such as poor mechanical strength, adhesion, etc. A hydrogel dressing with good hemostatic effect, good antibacterial performance and effective wound healing promotion is urgently needed. SUMMARY

[0004] An object of the present application is to provide a hydrogel dressing with high adhesion and strong antibacterial property, which is suitable for various traumatic wounds and has the effects of hemostasis, pain relief, reduction of wound infection and promotion of wound healing.

[0005] Another object of the present application is to provide a preparation method of the hydrogel dressing.

[0006] Still another object of the present application is to provide an application of the hydrogel dressing.

[0007] In order to achieve the above objects, the present application provides an oxidized konjac gum and hydrazide hyaluronic acid composite hydrogel, which comprises oxidized konjac gum (OKGM), hydrazide hyaluronic acid (ADHHA), neomycin sulfate (NS), cannabidiol (CBD) and 2-butanol (2B).

[0008] As a preferred embodiment, the oxidized konjac gum is an oxidized modified konjac gum. More preferably, the oxidized konjac gum is a product obtained by oxidizing konjac gum with an oxidizing agent (for example, sodium periodate).

[0009] As a preferred embodiment, the preparation steps of the oxidized konjac gum are as follows: konjac gum is dissolved in water to prepare a 1-5 wt% solution, an oxidizing agent is added under light protection, the concentration of the oxidizing agent in the reaction system is 0.1-1 wt%, stirring is continued for 12-24 hours, then a reaction termination agent is added, the concentration is 1-5 wt%, and stirring is continued for 0.5-2 hours, the reaction is terminated, dialysis is performed using a dialysis bag, and freeze-drying is performed to obtain the oxidized konjac gum.

[0010] As a preferred embodiment, the molecular weight of the konjac gum is 20-2 million.

[0011] As a preferred embodiment, the hydrazide-modified hyaluronic acid is adipic acid dihydrazide-modified hyaluronic acid.

[0012] As a preferred embodiment, the preparation steps of the hydrazide-modified hyaluronic acid are as follows: hyaluronic acid is dissolved in water to prepare a 1-5 wt% solution, then a hydrazide reagent and an activating agent are added, the concentration of the hydrazide reagent and the activating agent in the reaction system is both 0.1-1 wt%, and the pH is adjusted to 4.75, stirring is continued for 12-24 hours, dialysis is performed using a dialysis bag, and freeze-drying is performed to obtain the hydrazide-modified hyaluronic acid.

[0013] As a preferred embodiment, the molecular weight of the hyaluronic acid is 15-20 million.

[0014] The present application utilizes the Schiff base crosslinking of the oxidized konjac gum and the hydrazide-modified hyaluronic acid, and loads neomycin sulfate, to form a hydrogel dressing for wrapping cannabidiol and 2-CTS alcohol.

[0015] As a preferred embodiment, the mass ratio of the oxidized konjac gum to the hydrazide-modified hyaluronic acid is (1-5):(1-5), for example (1-3):(1-2).

[0016] As a preferred embodiment, the molar ratio of neomycin sulfate, cannabidiol, and 2-CTS alcohol is neomycin sulfate:cannabidiol:2-CTS alcohol = (1-5):(1-5):(1-5).

[0017] As a preferred embodiment, the oxidation degree of the oxidized konjac gum is 20%-60%.

[0018] As a preferred embodiment, the hydrazide degree of the hydrazide-modified hyaluronic acid is 10%-50%.

[0019] In another aspect, the present application also provides a preparation method of the oxidized konjac gum and the hydrazide-modified hyaluronic acid composite hydrogel, which comprises the following steps:

[0020] The oxidized konjac gum solution and the neomycin sulfate solution are mixed and reacted, then the cannabidiol solution, the 2-butanol solution are added in sequence, and finally the hydrazide hyaluronic acid solution is added, and the gel is formed after rapid stirring and standing.

[0021] The mass concentration of the oxidized konjac gum in the oxidized konjac gum solution is 1%-5%, and the solvent is water.

[0022] The mass concentration of the hydrazide hyaluronic acid in the hydrazide hyaluronic acid solution is 1%-5%, and the solvent is water.

[0023] The concentration of the neomycin sulfate in the neomycin sulfate solution is 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

[0024] The concentration of the cannabidiol in the cannabidiol solution is 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

[0025] The concentration of the 2-butanol in the 2-butanol solution is 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

[0026] The volume ratio of the oxidized konjac gum solution, the hydrazide hyaluronic acid solution, the neomycin sulfate solution, the cannabidiol solution and the 2-butanol solution in the hydrogel dressing is 1:1:0.01:0.01:0.01.

[0027] In another aspect, the application also provides the use of the composite hydrogel in the preparation of a hemostatic material.

[0028] Hyaluronic acid is a natural hyaluronic acid, which widely exists in human connective tissue, has high water retention, no immunity, can be degraded, and the product is non-toxic, has repair promoting effect and adhesion. Konjac gum is a natural konjac gum extracted from konjac tuber, which has high water absorption, biocompatibility, adhesion and other effects. Both have good biocompatibility, degradability and other characteristics, and are ideal components for preparing wound hemostatic and healing promoting hydrogel. The hydrogel dressing of the application has good hemostatic and antibacterial effects, can effectively promote wound healing, has simple preparation steps, natural polymer materials, non-toxic, easy to obtain, low price, and wide application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 The frequency scanning and amplitude scanning results of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to the embodiment of the application are shown. (A) Frequency scanning; (B) Amplitude scanning.

[0030] Figure 2The ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to an embodiment of the present invention is shown to inhibit Escherichia coli (E. coli) E. coli ) and Staphylococcus aureus ( S. aureus The inhibitory effect of ).

[0031] Figure 3 The results show the colony count and inhibition rate of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel against Escherichia coli and Staphylococcus aureus according to embodiments of the present invention. (A) Bacterial growth on agar plates; (B) Inhibition rate against bacteria.

[0032] Figure 4 The cell compatibility of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel at different concentrations according to embodiments of the present invention is shown.

[0033] Figure 5 The blood compatibility of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to an embodiment of the present invention is shown. (A) Hemolysis test of the hydrogel; (B) Hemolysis rate.

[0034] Figure 6 The coagulation index (BCI) of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to an embodiment of the present invention is shown. (A) BCI index of the hydrogel; (B) Coagulation properties of the hydrogel.

[0035] Figure 7 The hemostatic effect of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to an embodiment of the present invention on mouse liver is shown. (A) Hemostasis status of mouse liver; (B) Hemostasis time of mouse liver; (C) Amount of bleeding in mouse liver.

[0036] Figure 8 The effects and healing rates of the ADHHA / OKGM / NS / CBD / 2B composite hydrogel according to embodiments of the present invention on mouse wound healing are shown. (A) Changes in mouse wound healing over time after hydrogel application; (B) Wound healing rate on day 10 in mice. ***P≤0.001 indicates a significant difference. Detailed Implementation

[0037] To facilitate understanding of the present invention, a more comprehensive description will be provided below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0038] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0039] In this invention, oxidized konjac gum is preferably obtained by oxidizing the ortho-hydroxyl group of konjac gum to an aldehyde group. The oxidation degree of oxidized konjac gum is preferably 20-60%. For example, it can be 20%, 30%, 40%, 50%, 60%, etc. The molecular weight of konjac gum can be 200,000 to 2,000,000.

[0040] The hydrazide-modified hyaluronic acid of the present invention is preferably prepared by reacting the hydroxyl groups on the hyaluronic acid with the hydrazide-modifying reagent.

[0041] In this invention, the degree of hydrazideation of the hyaluronic acid is preferably 10-50%. For example, it can be 10%, 20%, 30%, 40%, 50%, etc. The molecular weight of the hyaluronic acid can be 150,000-200,000.

[0042] In the neomycin sulfate solution, the concentration of neomycin sulfate is preferably 0.5 mol / L to 1 mol / L, and the solvent is water.

[0043] In the cannabidiol solution, the concentration of cannabidiol is preferably 0.5 mol / L - 1 mol / L, and the solvent is anhydrous ethanol.

[0044] In the 2-Ciol solution, the concentration of 2-Ciol is 0.5 mol / L - 1 mol / L, and the solvent is anhydrous ethanol.

[0045] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but they should not be construed as limiting the scope of patent protection of the present invention.

[0046] Unless otherwise specified, the reagents, methods and equipment used in the implementation of this invention are conventional reagents, methods and equipment in the technical field.

[0047] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0048] Example 1

[0049] A novel hydrogel wound dressing is prepared according to the following steps:

[0050] (1) 5 g konjac glucomannan (KGM) was added into 500 mL deionized water and stirred until completely dissolved. Then, 1.32 g NaIO4 was added, and the mixture was stirred at 30 °C for 12 hours in the dark. Next, 10 mL ethylene glycol was added to terminate the reaction, and the mixture was stirred for another 2 hours. The resulting solution was then dialyzed in a dialysis bag in deionized water for 3 days, freeze-dried to obtain oxidized konjac glucomannan (OKGM). The oxidized konjac glucomannan was dissolved in deionized water at 50 °C to obtain a 5% oxidized konjac glucomannan solution (5 g oxidized konjac glucomannan: 100 mL deionized water).

[0051] (2) 5 mmol neomycin sulfate (NS) was weighed into a beaker, dissolved in 5 mL deionized water to obtain a neomycin sulfate solution (concentration of 1 mol / L).

[0052] (3) 2 mmol cannabidiol (CBD) was weighed into a vial, dissolved in anhydrous ethanol to 2 mL to obtain a cannabidiol solution (concentration of 1 mol / L).

[0053] (4) 2 mmol 2-azidoethanol (2B) was weighed into a vial, dissolved in anhydrous ethanol to 2 mL to obtain a 2-azidoethanol solution (concentration of 1 mol / L).

[0054] (5) 2 g hyaluronic acid (HA) was dissolved in 200 mL sterile water. 2.3 g ADH was added to the solution, and 1.84 g ADH and 1.43 g HOBT were slowly added to the HA solution drop by drop while stirring, and the pH was adjusted to 4.75; 0.91 g EDC was slowly added to the solution drop by drop while stirring, and the pH was adjusted with 1 M HCl and 1 M NaOH, and the pH was maintained at 4.75 for 4 hours; the solution was continuously stirred at room temperature for 24 hours, and then dialyzed in deionized water for 3 days, frozen at -80 °C, and freeze-dried to obtain hydrazide hyaluronic acid (ADHHA), which was proportionally configured into a 5% hydrazide hyaluronic acid solution.

[0055] (6) 1 mL oxidized konjac glucomannan solution (concentration of 5% w / v) was taken in a container, 10 μL neomycin sulfate solution (concentration of 1 mol / L) was added, and the solution was stirred until uniform; 10 μL cannabidiol solution (concentration of 1 mol / L) and 10 μL 2-azidoethanol solution (concentration of 1 mol / L) were added to the solution respectively, and the solution was stirred until uniform; finally, 1 mL hydrazide hyaluronic acid solution (concentration of 5% w / v) was added, and the solution was continuously stirred and then observed; the solution formed a hydrogel in 5 minutes.

[0056] Example 2

[0057] A novel hydrogel wound dressing is prepared in a way that differs from the preparation method in Example 1 only in that the concentration of oxidized konjac gum is changed; in this example, the concentration of oxidized konjac gum is 3%.

[0058] Example 3

[0059] A novel hydrogel wound dressing is prepared in a way that differs from the preparation method in Example 1 only in that the concentration of oxidized konjac gum is changed; in this example, the concentration of oxidized konjac gum is 2%.

[0060] Example 4

[0061] A novel method for preparing a hydrogel wound dressing includes the following steps:

[0062] (1) Add 5 g of konjac gum (KGM) to 500 mL of deionized water and stir until completely dissolved. Then, add 1.32 g of NaIO4 and stir the mixture at 30 °C in the dark for 12 hours. Next, add 10 mL of ethylene glycol to terminate the reaction and stir the mixture for another 2 hours. Then, dialyze the resulting solution in deionized water using a dialysis bag for 3 days and freeze-dry to obtain oxidized konjac gum (OKGM). Prepare a 3% oxidized konjac gum solution according to the ratio (3 g oxidized konjac gum: 100 mL deionized water).

[0063] (2) Weigh 5 mmol of neomycin sulfate into a beaker, add 5 mL of deionized water to dissolve it, and obtain a neomycin sulfate solution (concentration of 1 mol / L).

[0064] (3) Weigh 2 mmol of cannabidiol into a vial, add anhydrous ethanol to make up to 2 mL, and obtain a cannabidiol solution (concentration of 1 mol / L).

[0065] (4) Weigh 2 mmol of 2-Ciol into a vial, add anhydrous ethanol to make up to 2 mL, and obtain a 2-Ciol solution (concentration of 1 mol / L).

[0066] (5) 2 g of hyaluronic acid (HA) was dissolved in 200 mL of sterile water. 2.3 g of ADH was added to the solution, and 1.84 g of ADH and 1.43 g of HOBT were slowly added to the HA solution while stirring, and the pH was adjusted to 4.75; 0.91 g of EDC was slowly added to the solution, and the pH was adjusted with 1 M HCl and 1 M NaOH, and the pH was maintained at 4.75 for 4 hours, and then the reaction was continued at room temperature for 24 hours, and then the reaction was stopped by dialysis against deionized water for 3 days at -80°C, and then the hydrazide hyaluronic acid (ADHHA) was obtained by freeze-drying, and a 2% hydrazide hyaluronic acid solution was prepared according to the proportion.

[0067] (6) 1 mL of oxidized konjac gum solution (3% w / v) was taken in a container, 10 μL of neomycin sulfate solution (1 mol / L) was added, and the solution was stirred evenly, and then 10 μL of cannabidiol solution (1 mol / L) and 10 μL of 2-mercaptoethanol solution (1 mol / L) were added to the solution, and the solution was stirred evenly, and finally 1 mL of hydrazide hyaluronic acid solution (2% w / v) was added, and the solution was observed after continuous stirring. The solution forms a hydrogel in 3 minutes.

[0068] Example 5

[0069] A new type of hydrogel wound dressing, the only difference between the preparation method and the preparation method of Example 4 is to change the concentration of oxidized konjac gum; in this example, the concentration of oxidized konjac gum is 4%.

[0070] Example 6

[0071] A new type of hydrogel wound dressing, the only difference between the preparation method and the preparation method of Example 4 is to change the concentration of oxidized konjac gum; in this example, the concentration of oxidized konjac gum is 5%.

[0072] In the present application, the adjacent hydroxyl groups in the konjac gum structure can be oxidized to aldehyde groups, and the amino groups in the hydrazide hyaluronic acid derivative can undergo Schiff base reaction, and a dynamic Schiff base bond with reversible characteristics can be formed, and the hydrogen bond formed between the amino groups and the hydroxyl groups can strengthen the bonding degree of the three-dimensional network of the hydrogel, and a new type of hydrogel dressing with fast and strong self-healing properties is prepared.

[0073] Performance test

[0074] The antibacterial performance and biocompatibility of the hydrogel were tested according to GB / T 31402-2015 and GB / T 16886.5-2003, respectively.

[0075] Biotoxicity score: no cytotoxicity (0), slight cytotoxicity (1), moderate cytotoxicity (2), severe cytotoxicity (3).

[0076] The hemolysis rate is calculated as follows:

[0077] Hemolysis rate = (absorbance of experimental group - absorbance of negative control group) / (absorbance of positive control group - absorbance of negative control group) × 100%.

[0078] The gelation times of the hydrogel samples obtained in Examples 1-6 above are shown in Table 1.

[0079] Table 1: Comparison of hydrogel forming time in the examples

[0080]

[0081] As shown in Table 1, the gelation time of the hydrogel changes with the concentration and ratio of oxidized konjac gum and hydrazide-modified hyaluronic acid. The shortest gelation time, 10 seconds, is achieved when the ratio of oxidized konjac gum to hydrazide-modified hyaluronic acid is 2:1. This is of great significance for preparing rapid and self-healing Schiff base hydrogels.

[0082] The properties of the composite hydrogels in Examples 1-6 were measured. The results showed that the composite hydrogels in Examples 1-6 all had good hydrogel structures, significant antibacterial effects, were harmless and non-toxic to cells, had excellent blood compatibility, and could effectively coagulate, stop bleeding, and promote wound healing.

[0083] The following explanation uses the hydrogel performance test results from Example 5 as an example.

[0084] Rheological analysis was performed on the hydrogel of Example 5. The results are as follows: Figure 1 As shown, in Example 5, the hydrogel's G' is always greater than G' when the angular frequency is ≤20. Figure 1 The values ​​in A indicate that the gel remains hydrogel within this range, which represents the elastic range of the hydrogel. When the shear strain is ≤13, G' and G” do not change significantly with shear strain and can be considered constants. Figure 1 (B) proves that this region is the linear viscoelastic region of the hydrogel, and the hydrogel structure will not change due to strain.

[0085] As a wound dressing, antibacterial properties are essential; therefore, the hydrogel in Example 5 was tested for antibacterial activity. *Escherichia coli* and *Staphylococcus aureus* were selected as bacterial models, and its antibacterial properties were determined using the inhibition zone method. The results are as follows: Figure 2 As shown. Figure 2In the figure, section A represents a hydrogel composed solely of oxidized konjac gum and hydrazide-modified hyaluronic acid (the components of Example 5, omitting neomycin sulfate, cannabidiol, and 2-carboxylic acid), and section B represents the hydrogel of Example 5. As can be seen from the figure, after loading with neomycin sulfate, the hydrogel exhibits significant inhibitory effects against Staphylococcus aureus and Escherichia coli. Therefore, the hydrogel prepared in this invention possesses antibacterial properties.

[0086] The antibacterial rate was determined using the dilution plating method. The hydrogel and the bacterial suspension co-cultured with the two bacteria were diluted by the same factor, plated, and counted. The results are as follows: Figure 3 As shown in the figure, compared with the blank control, the inhibitory effect on Staphylococcus aureus and Escherichia coli is significant, with no bacterial growth on the culture medium and an inhibition rate of 100%. This result is consistent with the results of the inhibition zone experiment, indicating that the hydrogel prepared in this invention has an inhibitory effect on both bacteria.

[0087] Wound dressings must be non-toxic to humans and not cause adverse reactions after use. Therefore, 3T3 fibroblasts were selected, and the toxicity of hydrogels at different concentrations (0.625 mg / mL, 1.25 mg / mL, 2.5 mg / mL, and 5 mg / mL) to the cells was detected using the CCK8 assay. The results are as follows: Figure 4 As shown in the figure. The results indicate that, compared with the blank control, the cell viability of the hydrogel in Example 5 remained essentially unchanged with increasing concentration, remaining above 90%, except for a slight decrease in cell viability when the concentration reached 1 mg / mL.

[0088] The hydrogel prepared in Example 5 of this invention is crucial for blood compatibility, from Figure 5 As can be seen, the hydrogel in Example 5 exhibits significantly better blood compatibility compared to the control group (1.8 mL of sterile water was mixed with 36 µL of erythrocyte suspension as a positive control). Figure 5 (A). Furthermore, the hemolysis rate calculated by measuring ultraviolet absorbance was also below 5%, indicating that the hydrogel has excellent blood compatibility. Figure 5 B).

[0089] The most important characteristic of wound materials is their ability to aggregate platelets, thereby promoting blood clotting. Figure 6 In the hydrogel group, blood can be clearly seen coagulating at the bottom to form a dark red blood clot, proving that the hydrogel has a very good coagulation effect. Figure 6 B). By Figure 6 It can be seen that their coagulation index (BCI) is all below 40%, indicating that the hydrogel has a significant coagulation effect. Figure 6 A).

[0090] The Blood Clotting Index (BCI) is calculated using the following formula:

[0091]

[0092] The OD value is the OD value at 540 nm measured by an enzyme-linked immunosorbent assay (ELISA) reader.

[0093] The lower the BCI, the more effectively blood clotting can be stimulated.

[0094] A hemorrhage effect was induced using a mouse liver model, and then hydrogel adhesion was used to achieve the effect for quantitative analysis. Experimental results are as follows: Figure 7 As shown. From Figure 7 It can be seen that when the mouse liver is cut open without any treatment, the amount of bleeding is relatively large, reaching 0.2081g. However, when hydrogel is applied, the amount of bleeding is greatly reduced to only about 0.04g. Figure 7 (A, C). The bleeding time is also greatly shortened, stopping in about 32 seconds. Figure 7 Therefore, the hydrogel of the present invention has a very obvious effect on wound hemostasis, and when the liver of mice was cleaned, obvious blood clots were found to be formed at the bleeding site, indicating that the hydrogel achieves its effect by promoting coagulation.

[0095] In mouse wound healing experiments, from Figure 8 It was found that when only 3M dressing was applied, white purulent discharge appeared on the mouse wounds, and this discharge persisted from the third day to the tenth day. When hydrogel was applied inside the 3M dressing, no purulent discharge appeared on the mouse wounds, and the wounds gradually healed, showing significantly better healing by the tenth day. Figure 8 (A), the healing rate reaches approximately 98% ( Figure 8 B).

Claims

1. A composite hydrogel of oxidized konjac gum and hydrazide-modified hyaluronic acid for hemostasis, characterized in that... Including oxidized konjac gum, hydrazide-modified hyaluronic acid, neomycin sulfate, cannabidiol, and 2-tscissorcinol; The mass ratio of oxidized konjac gum to hydrazide-modified hyaluronic acid is (1-5):(1-5); The molar ratio of neomycin sulfate, cannabidiol, and 2-Citrol is (1-5):(1-5):(1-5).

2. The preparation method of the oxidized konjac gum and hydrazide-modified hyaluronic acid composite hydrogel for hemostasis as described in claim 1, characterized in that, The method includes the following steps: First, thoroughly mix the oxidized konjac gum solution with the neomycin sulfate solution to allow the reaction to proceed. Then, add the cannabidiol solution and 2-carboxylic acid solution in sequence, and finally add the hydrazide-modified hyaluronic acid solution. Stir rapidly and let stand to form the gum.

3. The method according to claim 2, characterized in that, The oxidized konjac gum solution has a mass-volume concentration of 1%-5% and uses water as the solvent.

4. The method according to claim 2, characterized in that, The hydrazide-modified hyaluronic acid solution has a mass-volume concentration of 1%-5% and uses water as the solvent.

5. The method according to claim 2, characterized in that, The concentration of neomycin sulfate in the solution is 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

6. The method according to claim 2, characterized in that, The cannabidiol solution contains cannabidiol at a concentration of 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

7. The method according to claim 2, characterized in that, In the 2-Ciol solution, the concentration of 2-Ciol is from 0.5 mol / L to 1 mol / L, and the solvent is anhydrous ethanol.

8. The use of the oxidized konjac gum and hydrazide-modified hyaluronic acid composite hydrogel as described in claim 1 in the preparation of hemostatic materials.

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