Adaptive drug-loaded hydrogel and preparation method and application thereof

Abstract

The invention discloses an adaptive drug-loaded hydrogel and a preparation method and application thereof, belonging to the technical field of biomedical materials. The adaptive hydrogel of the invention uses aldehyde-modified γ-cyclodextrin, sodium hyaluronate grafted coumarin and sodium hyaluronate grafted adipic acid dihydrazide as carrier raw materials, and antibacterial agents and insulin substances are loaded on the carrier, and the adaptive drug-loaded hydrogel is prepared by the host-guest complexation of γ-cyclodextrin and coumarin and the hydrazone bond series action of the aldehyde group of aldehyde-modified γ-cyclodextrin and the hydrazide group of sodium hyaluronate grafted adipic acid dihydrazide. The adaptive hydrogel drug-loaded hydrogel can not only rearrange the gel network, effectively fill and match the wound surface of complex shape, but also can release levofloxacin and insulin for a long time at the wound surface, play the role of inhibiting bacterial infection, promoting endothelial cell proliferation, angiogenesis and collagen deposition, thereby significantly accelerating the healing process of diabetic infected wounds.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to an adaptive drug-loaded hydrogel and a preparation method and application thereof. Background Art

[0002] Diabetes is a highly prevalent chronic metabolic disease worldwide. It often causes a series of complications during the development of diabetes. Among them, 15% to 25% of diabetic patients may suffer from diabetic foot ulcers caused by factors such as lower limb peripheral neuropathy and wound infection, which are recorded as DFUs. Under normal circumstances, the repair of skin wounds goes through four stages: hemostasis, inflammation, proliferation and differentiation, and tissue remodeling. Any interference in any stage of this complex physiological process will delay wound healing. However, the immune capacity of diabetic infected wounds is impaired under hyperglycemia. The continuous high-level expression of inflammatory factors and bacterial microbial infection seriously hinder wound healing. In extreme cases, gangrene may even lead to amputation or even death. Therefore, reshaping the wound microenvironment and inhibiting bacterial infection are crucial for the healing of diabetic infected wounds.

[0003] At present, traditional medical dressings used for the treatment of diabetic infected wounds include medical gauze, bandages and sponges. Although traditional dressings can absorb wound exudate, keep wounds dry and provide a certain degree of protection, they cannot effectively inhibit bacterial proliferation, cannot meet the needs of moist wound healing, and may adhere to the wound, causing secondary damage to the wound when replaced. Hydrogel is a cross-linked polymer material with high water content and high porosity. It has many properties similar to the natural extracellular matrix and can provide a moist environment for cell proliferation and tissue repair. In addition, it also has mechanical properties that match tissues, good biosafety and drug loading and release properties, and can be used as a functional dressing for the healing of chronic diabetic wounds.

[0004] Ideal diabetic chronic wound dressings should not only be able to reshape the wound microenvironment through mechanisms such as hypoglycemia and antibacterial, but also be able to effectively fill complex wounds and match the shape changes during wound healing. However, most hydrogel dressings are usually composed of static polymer networks, which not only lack tissue adhesion, but also have difficulty in reshaping the network to adapt to the shape of the wound, resulting in low drug delivery efficiency and unsatisfactory treatment effects. Summary of the invention

[0005] The present invention provides an adaptive drug-loaded hydrogel and a preparation method and application thereof, which effectively solve the technical problems that the existing hydrogel dressings are composed of static polymer networks, which not only lack tissue adhesion, but also have difficulty in reshaping the network to adapt to the shape of the wound, resulting in low drug delivery efficiency and unsatisfactory treatment effect; at the same time, a drug-loaded hydrogel dressing with excellent adaptability, adhesion, antibacterial properties, biocompatibility and the ability to promote rapid healing of diabetic infected wounds is constructed.

[0006] The present invention provides a method for preparing an adaptive drug-loaded hydrogel, comprising the following steps:

[0007] Dissolving aldehyde-modified γ-cyclodextrin and an antibacterial agent in water, stirring at room temperature in the dark to obtain a crude product, dispersing the crude product in an organic solvent, filtering to remove the antibacterial agent not included by the aldehyde-modified γ-cyclodextrin to obtain insoluble matter, and drying to obtain an aldehyde-modified γ-cyclodextrin / antibacterial agent inclusion complex;

[0008] The aqueous solution of the aldehyde-modified γ-cyclodextrin / antimicrobial agent inclusion complex is mixed with the aqueous solution of sodium hyaluronate grafted coumarin, and the γ-cyclodextrin and coumarin produce a host-guest complexation to form a host-guest assembly solution containing the antimicrobial agent. Then, an insulin solution and a sodium hyaluronate grafted adipic acid dihydrazide solution are added, and the mixture is incubated at 20-37° C. The aldehyde group of the host-guest assembly containing the antimicrobial agent undergoes a hydrazone bond coupling reaction with the hydrazide group of the sodium hyaluronate grafted adipic acid dihydrazide to obtain a hydrogel carrier, and the antimicrobial agent and insulin are loaded on the carrier at the same time to obtain an adaptive drug-loaded hydrogel.

[0009] As a preferred embodiment, the concentration of the antibacterial agent in the adaptive drug-loaded hydrogel is 1.10 mg / mL to 6.59 mg / mL.

[0010] As a preferred embodiment, the concentration of the insulin solution in the adaptive drug-loaded hydrogel is 0.1 U / mL~1 U / mL.

[0011] As a preferred embodiment, the antibacterial agent is levofloxacin, curcumin, ciprofloxacin, tetracycline or baicalin.

[0012] As a preferred embodiment, the molar ratio of γ-cyclodextrin to coumarin is 1:0.8-1:3, and the molar ratio of aldehyde group to hydrazide group is 1:0.5-1:2.

[0013] As a preferred embodiment, the molar ratio of γ-cyclodextrin to coumarin is 1:2, and the molar ratio of aldehyde group to hydrazide group is 1:1.

[0014] As a preferred embodiment, the solid content of the adaptive drug-loaded hydrogel is 5 w / v%~15 w / v%.

[0015] As a preferred embodiment, based on the number of aldehyde groups on each of the aldehyded γ-cyclodextrin molecules, the degree of substitution DSCHO of the aldehyded γ-cyclodextrin is 0.85~1.72; based on the number of moles of coumarin groups per mole of disaccharide unit, the degree of substitution DSCou of the sodium hyaluronate grafted coumarin is 0.15~0.60; based on the number of moles of adipic acid dihydrazide groups per mole of disaccharide repeating unit, the degree of substitution DSADH of the sodium hyaluronate grafted adipic acid dihydrazide is 0.15~0.55.

[0016] The second object of the present invention is to provide an adaptive drug-loaded hydrogel prepared by the above preparation method.

[0017] The third object of the present invention is to provide a use of the above-mentioned adaptive drug-loaded hydrogel in the preparation of a drug for treating wound infection caused by diabetes.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The invention provides an adaptive drug-loaded hydrogel and a preparation method and application thereof. The adaptive drug-loaded hydrogel of the invention uses aldehyded γ-cyclodextrin, denoted as γ-CD-CHO, sodium hyaluronate grafted coumarin, denoted as HA-Cou, and sodium hyaluronate grafted adipic acid dihydrazide, denoted as HA-ADH as carrier raw materials, and an antibacterial agent and an insulin substance are loaded on the carrier. The adaptive drug-loaded hydrogel is prepared through the host-guest complexation of γ-cyclodextrin and coumarin and the hydrazone bond series action of the aldehyde group of the aldehyded γ-cyclodextrin and the hydrazide group of the sodium hyaluronate grafted adipic acid dihydrazide. The adaptive drug-loaded hydrogel dressing prepared by the present invention has better adaptability than traditional hydrogel dressings. It can not only rearrange the gel network, effectively fill and match complex-shaped wounds, and increase the delivery efficiency of drugs to the wound, but also achieve efficient loading and long-term sustained release of antibacterial agents and insulin at the wound, play the role of inhibiting bacterial infection, promoting endothelial cell proliferation, angiogenesis and collagen deposition, thereby significantly accelerating the healing process of diabetic infection-type wounds. The adaptive drug-loaded hydrogel prepared by the present invention can achieve long-term antibacterial effect on the wound and accelerate wound recovery, and has great potential for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 : is the H NMR spectrum of γCD-COOH, HA-Cou and HA-ADH in the present invention, and the deuterated solvent is D2O.

[0021] Figure 2 Schematic diagram of the formation mechanism of the adaptive drug-loaded hydrogel in Example 1 of the present invention.

[0022] Figure 3Schematic diagram of the preparation of γCD-CHO@LOF prepared in Example 1 of the present invention.

[0023] Figure 4 It is the standard curve of LOF in Example 1 of the present invention.

[0024] Figure 5 This is a flow chart of the preparation of the adaptive drug-loaded hydrogel dressing in Example 1 of the present invention.

[0025] Figure 6 This is a flow chart of obtaining the adaptive drug-loaded hydrogel from the component solutions in Example 1 of the present invention.

[0026] Figure 7 This is a test diagram of the adhesion of the adaptive hydrogel dressing prepared in Example 1 of the present invention.

[0027] Figure 8 This is a self-healing test diagram of the adaptive hydrogel dressing prepared in Example 1 of the present invention.

[0028] Fig. 9 This is an adaptability test diagram of the adaptive hydrogel dressing prepared in Example 1 of the present invention.

[0029] Fig.10 The blood compatibility evaluation results of the hydrogel dressing prepared in the present invention are shown in FIG.

[0030] Fig.11 The results of hemolysis rate determination of the hydrogel dressing prepared in the present invention are shown in Figure 1. Gel is the blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is the adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is the adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is the adaptive drug-loaded hydrogel dressing in Example 1.

[0031] Fig.12 The cytotoxicity test results of the hydrogel dressing prepared in the present invention are shown in Table 1. Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1.

[0032] Fig.13The live / dead cell double staining evaluation results of the hydrogel dressing prepared in the present invention. Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1.

[0033] Fig.14 The results are the inhibition rates of the hydrogel dressings prepared in the present invention on Escherichia coli and Staphylococcus aureus. Wherein Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1. Compared with the blank control, *P<0.05, **P<0.01, ***P<0.001.

[0034] Fig.15 The results of the plate coating of the hydrogel dressing prepared in the present invention on Escherichia coli and Staphylococcus aureus. Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1.

[0035] Fig.16 The figure is a view of the healing process of the hydrogel dressing prepared in the present invention promoting the healing of diabetic infection-type wounds in mice. Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1.

[0036] Fig.17 The hydrogel dressing prepared in the present invention promotes the wound healing rate of mice in the diabetic infection-type wound healing experiment. Wherein Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1. Compared with the blank control, *P<0.05, ***P<0.001.

[0037] Fig.18 The hydrogel dressing prepared in the present invention promotes the change in the body weight of mice in the diabetic infection wound healing experiment. Control is a blank control, Gel is a blank adaptive hydrogel dressing in Comparative Example 1, Gel@LOF is an adaptive hydrogel dressing loaded with levofloxacin in Comparative Example 2, Gel@INS is an adaptive hydrogel dressing loaded with insulin in Comparative Example 3, and Gel@LOF / INS is an adaptive drug-loaded hydrogel dressing in Example 1. DETAILED DESCRIPTION

[0038] In order to enable those skilled in the art to better understand the technical solution of the present invention and implement it, the present invention is further described below in conjunction with specific embodiments and drawings, but the embodiments are not intended to limit the present invention. The following test methods and detection methods, unless otherwise specified, are conventional methods; the reagents and raw materials, unless otherwise specified, are commercially available.

[0039] Regarding the background technology mentioned in this application: Traditional medical dressings currently used for the treatment of diabetic infected wounds include medical gauze, bandages and sponges. Although traditional dressings can absorb wound exudate, keep wounds dry and provide a certain degree of protection, they cannot effectively inhibit bacterial proliferation, cannot meet the needs of moist wound healing, and may adhere to the wound, causing secondary damage to the wound when replaced. Most hydrogel dressings are usually composed of static polymer networks, which not only lack tissue adhesion, but also have difficulty in reshaping the network to adapt to the shape of the wound, resulting in low drug administration efficiency and unsatisfactory treatment effect. To this end, the present invention provides an adaptive drug-loaded hydrogel and its preparation method and application.

[0040] The technical solution of the present invention is described in detail below.

[0041] The present invention first provides a method for preparing an adaptive drug-loaded hydrogel, comprising the following steps:

[0042] The aldehyde-modified γ-cyclodextrin and the antimicrobial agent are dissolved in water, stirred at room temperature in the dark, and a crude product is obtained. The crude product is dispersed in an organic solvent, filtered, and the antimicrobial agent not included in the aldehyde-modified γ-cyclodextrin is removed to obtain an insoluble matter, which is dried to obtain an aldehyde-modified γ-cyclodextrin / antimicrobial agent inclusion complex, which is recorded as γCD-CHO@LOF.

[0043] The aqueous solution of the aldehyde-modified γ-cyclodextrin / antimicrobial agent inclusion complex is mixed with the aqueous solution of sodium hyaluronate grafted coumarin, and the γ-cyclodextrin and coumarin produce a host-guest complexation to form a host-guest assembly solution containing the antimicrobial agent. Then, an insulin solution and a sodium hyaluronate grafted adipic acid dihydrazide solution are added, and the mixture is incubated at 20° C. to 37° C. The aldehyde group of the host-guest assembly containing the antimicrobial agent undergoes a hydrazone bond coupling reaction with the hydrazide group of the sodium hyaluronate grafted adipic acid dihydrazide to obtain a hydrogel carrier, and the antimicrobial agent and insulin are loaded on the carrier at the same time to obtain an adaptive drug-loaded hydrogel.

[0044] The adaptive drug-loaded hydrogel dressing prepared by the above technical scheme has better adaptability than traditional hydrogel dressings. It can not only rearrange the gel network, effectively fill and match complex-shaped wounds, and increase the efficiency of drug delivery to the wound, but also achieve efficient loading of antibacterial agents and insulin and long-term sustained release at the wound, inhibiting bacterial infection, promoting endothelial cell proliferation, angiogenesis and collagen deposition, thereby significantly accelerating the healing process of diabetic infected wounds.

[0045] In order to ensure the therapeutic effect of the adaptive drug-loaded hydrogel on diabetic infected wounds, the concentration of the antibacterial agent in the adaptive drug-loaded hydrogel is 1.10 mg / mL to 6.59 mg / mL. If the amount of the antibacterial agent is less than 1.10 mg / mL, the antibacterial effect cannot be achieved. If the concentration of the antibacterial agent is greater than 6.59 mg / mL, it will have a certain toxicity to cells.

[0046] In order to improve the healing rate of infected wounds, the concentration of insulin in the adaptive drug-loaded hydrogel is 0.1U / mL~1 U / mL. Similarly, when the insulin concentration is too low, less than 0.1 U / mL, the wound healing effect is not ideal. When the insulin concentration is too high, greater than 1 U / mL, saturation occurs, resulting in drug waste.

[0047] In order to promote wound healing of wounds infected with different bacterial types, the antibacterial agent is levofloxacin, curcumin, ciprofloxacin, tetracycline or baicalin.

[0048] In order to obtain an adaptive drug-loaded hydrogel with different drug loading and release properties and mechanical properties, the molar ratio of γ-cyclodextrin to coumarin is 1:0.8~1:3, and the molar ratio of aldehyde group to hydrazide group is 1:0.5~1:2. More preferably, the molar ratio of γ-cyclodextrin to coumarin is 1:2, and the molar ratio of aldehyde group to hydrazide group is 1:1. When γ-cyclodextrin and coumarin are included in a molar ratio of 1:2, the aldehyde group on the γ-cyclodextrin and the hydrazide group on the adipic acid dihydrazide react in a 1:1 ratio. This ratio will make the hydrogel produce the best cross-linking state and the best performance.

[0049] In order to further obtain a hydrogel in an optimal state, the solid content of the adaptive drug-loaded hydrogel is 5 w / v%~15 w / v%. When the solid content is too low, less than 5 w / v%, the hydrogel cannot be formed. When the solid content is too high, greater than 15 w / v%, sodium hyaluronate grafted coumarin and sodium hyaluronate grafted adipic acid dihydrazide are difficult to dissolve in water, and an aqueous solution cannot be obtained, thereby affecting the formation of the hydrogel.

[0050] In order to obtain an adaptive drug-loaded hydrogel with better performance, the degree of substitution DSCHO of the aldehyded γ-cyclodextrin is 0.85-1.72, based on the number of aldehyde groups on each aldehyded γ-cyclodextrin molecule; the degree of substitution DSCou of the sodium hyaluronate grafted coumarin is 0.15-0.60, based on the number of moles of coumarin groups per mole of disaccharide unit; and the degree of substitution DSADH of the sodium hyaluronate grafted adipic acid dihydrazide is 0.15-0.55, based on the number of moles of adipic acid dihydrazide groups per mole of disaccharide repeating unit.

[0051] The technical effects of the present invention are described below through specific embodiments and comparative examples.

[0052] The present invention first prepares aldehyde-modified γ-cyclodextrin, denoted as γCD-CHO, sodium hyaluronate grafted with coumarin, denoted as HA-Cou, and sodium hyaluronate grafted with adipic acid dihydrazide, denoted as HA-ADH, as follows:

[0053] 1. Preparation of γCD-CHO: γCD, 500 mg, 0.385 mmol disaccharide unit and Dess-Martin reagent, 408.8 mg, 0.97 mmol were weighed and dissolved in 20 mL of ultra-dry dimethyl sulfoxide. After stirring and reacting at room temperature for 3 hours, the reaction solution was added dropwise to 100 mL of cold acetone for reprecipitation. After centrifugation to obtain the precipitate, 2 mL of water was added to dissolve it, and it was added dropwise to 20 mL of cold acetone again for reprecipitation. The precipitate was collected by centrifugation and dried in vacuo at 35°C to obtain the product γCD-CHO, whose structural formula is: .

[0054] In order to determine the degree of substitution of γCD-CHO, γCD-CHO was derivatized with hydroxylamine compounds to obtain carboxylated γCD, which was recorded as γCD-COOH. The specific steps are as follows: γCD-CHO, 100 mg, 0.08 mmol, and carboxymethyloxyamine hemihydrochloride, 41.7 mg, 0.38 mmol, were weighed and dissolved in 10 mL of deionized water, 0.5 M NaOH was used to adjust the solution pH to 4.8, and the reaction was stirred at room temperature for 48 h, and then the pH was adjusted to 7 with 0.5 M NaOH, and the reaction solution was added dropwise to 50 mL of cold acetone for reprecipitation, and the precipitate was obtained by centrifugation and 1 mL of water was added to dissolve it, and it was added dropwise to 10 mL of cold acetone again for reprecipitation, and the precipitate was collected by centrifugation and dried in vacuo at 35°C to obtain the product γCD-COOH, whose structural formula is:

[0055] , its H NMR spectrum, the deuterated solvent is D2O, see Figure 1 Figure a in the figure shows that DSCHO=1.0 as confirmed by NMR.

[0056] 2. Preparation of HA-ADH: Sodium hyaluronate, 500 mg, 1.24 mmol disaccharide unit, and adipic acid dihydrazide, 1085 mg, 6.23 mmol, were dissolved in 10 mM, 100 mL 4-morpholineethanesulfonic acid aqueous solution, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 83.59 mg, 0.44 mmol, and N-hydroxysuccinimide, 50.18 mg, 0.44 mmol, were added, stirred and reacted at room temperature in the dark for 24 hours, dialyzed against deionized water using a dialysis bag with a molecular weight cutoff of 8000-14000Da, and finally the product HA-ADH was obtained after freeze drying, and its structural formula is:

[0057] , its H NMR spectrum, the deuterated solvent is D2O, see Figure 1 In Figure b, DSADH=0.3 was confirmed by NMR.

[0058] 3. Preparation of HA-Cou: Weigh 500 mg of sodium hyaluronate and 1.24 mmol of disaccharide units, dissolve in 25 mL of deionized water, add 2500 mg of 732 hydrogen-type ion exchange resin, stir and react at room temperature for 4 hours, collect the supernatant solution by centrifugation, adjust the pH of the aqueous solution to 7 with 25% tetrabutylammonium hydroxide, denoted as TBA-OH, and freeze-dry to obtain hyaluronic acid tetrabutylammonium salt. Weigh 500 mg of hyaluronic acid tetrabutylammonium salt, 0.81 mmol of disaccharide unit and 532 mg of carboxymethyl coumarin, 2.4 mmol, and dissolve in 20 mL of ultra-dry dimethyl sulfoxide. Then add 98.4 mg of 4-dimethylaminopyridine, 0.81 mmol, and 185 μL of di-tert-butyl dicarbonate, 0.81 mmol. Stir and react in an oil bath at 45°C in the dark for 24 hours. Use a dialysis bag with a molecular weight cutoff of 8000-14000Da to dialyze dimethyl sulfoxide, 1M NaCl solution and deionized water in turn. After freeze-drying, the product HA-Cou is finally obtained, and its structural formula is: , its H NMR spectrum, the deuterated solvent is D2O, see Figure 1 Figure c in the figure shows that DSCou=0.3 as confirmed by NMR.

[0059] Example 1

[0060] A method for preparing an adaptive drug-loaded hydrogel, the formation mechanism of which is as follows Figure 2 The preparation process is as shown in Figure 5 As shown, the following steps are included:

[0061] S1. Prepare solutions of HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3 respectively: in a clean bench, weigh HA-Cou and HA-ADH respectively in sterile containers, add water or PBS buffer and vortex to dissolve, to prepare HA-Cou and HA-ADH solutions with a concentration of 10wt% each;

[0062] S2. Preparation of γCD-CHO@LOF: Dissolve γCD-CHO, DSCHO=1.0, 100 mg, 0.07 mmol, and LOF, 27.58 mg, 0.07 mmol, in 10 mL of deionized water, stir in the dark at room temperature for 48 h, and collect the product after freeze drying; then disperse the crude product in 5 mL of chloroform, collect the insoluble matter by filtration, and obtain γCD-CHO@LOF after vacuum drying; the schematic diagram of the preparation of γCD-CHO@LOF is shown in Figure 3 shown.

[0063] Dissolve 10 mg LOF in 10 mL PBS buffer solution to prepare a 1 mg / mL LOF solution, and dilute it by half with PBS buffer solution to prepare LOF solutions with different concentrations. Use an ELISA reader to measure the absorbance at 294 nm to establish a standard curve. Figure 4 .

[0064] The absorbance of the γCD-CHO@LOF solution was measured, and the LOF encapsulation efficiency was calculated to be 82.44% and the drug loading efficiency was 19.37% based on the above standard curve.

[0065] S3. Preparation of γCD-CHO@LOF solution: In a clean bench, weigh γCD-CHO@LOF into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a γCD-CHO@LOF solution with a concentration of 10 wt%;

[0066] S4. Prepare insulin solution, denoted as INS: In a clean bench, weigh INS into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a 10 U / mL INS solution;

[0067] S5. Prepare an adaptive drug-loaded hydrogel dressing: mix 100 μL of the HA-Cou solution prepared in step S1 and 63 μL of the γCD-CHO@LOF solution prepared in step S3 to form a host-guest assembly solution; then add 2.8 μL of the INS solution prepared in step S4 and 112 μL of the HA-ADH solution prepared in step S1 in sequence, stir evenly and incubate at room temperature or 37°C to obtain an adaptive drug-loaded hydrogel dressing through host-guest complexation and hydrazone bond tandem action, denoted as Gel@LOF / INS; its storage modulus is 513.80 Pa, and the concentrations of levofloxacin, denoted as LOF, and INS are 4.39 mg / mL and 0.1 U / mL, respectively.

[0068] Example 2

[0069] A method for preparing an adaptive drug-loaded hydrogel comprises the following steps:

[0070] S1. Prepare solutions of HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3: In a clean bench, weigh HA-Cou and HA-ADH respectively in sterile containers, add water or PBS buffer and vortex to dissolve, to prepare HA-Cou and HA-ADH solutions with a concentration of 10 wt% each;

[0071] S2. Preparation of γCD-CHO@LOF: Dissolve γCD-CHO, DSCHO=1.0, 100 mg, 0.07 mmol and LOF, 27.58 mg, 0.07 mmol in 10 mL of deionized water, stir in the dark at room temperature for 48 h, and collect the product after freeze-drying. Then, disperse the crude product in 5 mL of chloroform, collect the insoluble matter by filtration, and obtain γCD-CHO@LOF after vacuum drying.

[0072] 10 mg LOF was dissolved in 10 mL PBS buffer solution to prepare a 1 mg / mL LOF solution, which was then diluted by half with PBS buffer solution to prepare LOF solutions with different concentrations. The absorbance at a wavelength of 294 nm was measured using an enzyme marker to establish a standard curve.

[0073] The absorbance of the γCD-CHO@LOF solution was measured, and the LOF encapsulation efficiency was calculated to be 82.44% and the drug loading efficiency was 19.37% based on the above standard curve.

[0074] S3. Preparation of γCD-CHO@LOF and γCD-CHO solutions: In a clean bench, weigh γCD-CHO@LOF and γCD-CHO in sterile containers, add water or PBS buffer and vortex to dissolve, to prepare γCD-CHO@LOF and γCD-CHO solutions with a concentration of 10 wt%;

[0075] S4. Prepare insulin solution: In a clean bench, weigh INS into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a 1000 U / mL INS solution;

[0076] S5. Prepare an adaptive drug-loaded hydrogel dressing: mix 100 μL of the HA-Cou solution prepared in step S1, 31.5 μL of the γCD-CHO@LOF solution prepared in step S3, and 31.5 μL of the γCD-CHO solution to form a host-guest assembly solution; then add 2.8 μL of the INS solution prepared in step S3 and 112 μL of the HA-ADH solution prepared in step S1 in sequence, stir well, and incubate at room temperature or 37°C to obtain an adaptive drug-loaded hydrogel dressing through host-guest complexation and hydrazone bond tandem action; its storage modulus is 554.80 Pa, and the concentrations of LOF and INS are 2.49 mg / mL and 1 U / mL, respectively.

[0077] Example 3

[0078] A method for preparing an adaptive drug-loaded hydrogel comprises the following steps:

[0079] S1. Preparation of HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3 solutions: In a clean bench, weigh HA-Cou and HA-ADH respectively into sterile containers, add water or PBS buffer and vortex to dissolve, to prepare HA-Cou and HA-ADH solutions with a concentration of 5 wt% each;

[0080] S2. Preparation of γCD-CHO@LOF: Dissolve γCD-CHO, DSCHO=1.0, 100 mg, 0.07 mmol and LOF, 27.58 mg, 0.07 mmol, in 10 mL of deionized water, stir in the dark at room temperature for 48 h, and collect the product after freeze drying. Then, disperse the crude product in 5 mL of chloroform, collect the insoluble matter by filtration, and obtain γCD-CHO@LOF after vacuum drying.

[0081] 10 mg LOF was dissolved in 10 mL PBS buffer solution to prepare a 1 mg / mL LOF solution, which was then diluted by half with PBS buffer solution to prepare LOF solutions with different concentrations. The absorbance at a wavelength of 294 nm was measured using an enzyme marker to establish a standard curve.

[0082] The absorbance of the γCD-CHO@LOF solution was measured, and the LOF encapsulation efficiency was calculated to be 82.44% and the drug loading efficiency was 19.37% based on the above standard curve.

[0083] S3. Preparation of γCD-CHO@LOF solution: In a clean bench, weigh γCD-CHO@LOF into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a γCD-CHO@LOF solution with a concentration of 5 wt%;

[0084] S4. Prepare insulin solution: In a clean bench, weigh INS into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a 100 U / mL INS solution;

[0085] S5. Prepare an adaptive drug-loaded hydrogel dressing: mix 100 μL of the HA-Cou solution prepared in step S1 and 126 μL of the γCD-CHO@LOF solution prepared in step S3 to form a host-guest assembly solution; then add 2.8 μL of the INS solution prepared in step S4 and 112 μL of the HA-ADH solution prepared in step S1 in sequence, stir evenly, and incubate at room temperature or 37°C to obtain an adaptive drug-loaded hydrogel dressing through host-guest complexation and hydrazone bond tandem action; its storage modulus is 260.60 Pa, and the concentrations of LOF and INS are 4.39 mg / mL and 0.1 U / mL, respectively.

[0086] Example 4

[0087] A method for preparing an adaptive drug-loaded hydrogel comprises the following steps:

[0088] S1. Preparation of HA-Cou, DSCou=0.5 and HA-ADH, DSADH=0.37 solutions: In a clean bench, weigh HA-Cou with a degree of substitution of 0.5 and HA-ADH with a degree of substitution of 0.37 into sterile containers, add water or PBS buffer and vortex to dissolve, to prepare HA-Cou and HA-ADH solutions with a concentration of 10 wt% each;

[0089] S2. Preparation of γCD-CHO@LOF: Dissolve γCD-CHO, DSCHO=1.5, 100 mg, 0.07 mmol and LOF, 27.58 mg, 0.07 mmol in 10 mL of deionized water, stir in the dark at room temperature for 48 h, and collect the product after freeze drying. Then, disperse the crude product in 5 mL of chloroform, collect the insoluble matter by filtration, and obtain γCD-CHO@LOF after vacuum drying.

[0090] 10 mg LOF was dissolved in 10 mL PBS buffer solution to prepare a 1 mg / mL LOF solution, which was then diluted by half with PBS buffer solution to prepare LOF solutions with different concentrations. The absorbance at a wavelength of 294 nm was measured using an enzyme marker to establish a standard curve.

[0091] The absorbance of the γCD-CHO@LOF solution was measured, and the LOF encapsulation efficiency was calculated to be 82.44% and the drug loading efficiency was 19.37% based on the above standard curve.

[0092] S3. Preparation of γCD-CHO@LOF solution: In a clean bench, weigh γCD-CHO@LOF into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a γCD-CHO@LOF solution with a concentration of 10 wt%;

[0093] S4. Prepare insulin solution: In a clean bench, weigh INS into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a 100 U / mL INS solution;

[0094] S5. Prepare an adaptive drug-loaded hydrogel dressing: mix 100 μL of the HA-Cou solution prepared in step S1 and 63 μL of the γCD-CHO@LOF solution prepared in step S3 to form a host-guest assembly solution; then add 2.8 μL of the INS solution prepared in step S4 and 112 μL of the HA-ADH solution prepared in step S1 in sequence, stir evenly and incubate at room temperature or 37°C to obtain an adaptive drug-loaded hydrogel dressing through host-guest complexation and hydrazone bond tandem action; its storage modulus is 721.60 Pa, and the concentrations of LOF and INS are 4.39 mg / mL and 0.1 U / mL, respectively.

[0095] In order to further illustrate the technical effect of the present invention, the present invention also sets a comparative example, as follows:

[0096] Comparative Example 1, blank adaptive hydrogel

[0097] A method for preparing an adaptive hydrogel comprises the following steps:

[0098] S1. Preparation of γCD-CHO, DSCHO=1.0, HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3 solutions: In a clean bench, weigh γCD-CHO, HA-Cou and HA-ADH respectively in sterile containers, add water or PBS buffer and vortex to dissolve, to prepare γCD-CHO, HA-Cou and HA-ADH solutions with a concentration of 10 wt% each;

[0099] S2. Prepare a blank adaptive hydrogel dressing: mix 100 μL of the HA-Cou solution prepared in step S1 and 63 μL of the γCD-CHO solution to form a host-guest assembly solution; then add 112 μL of the HA-ADH solution, stir evenly and incubate at room temperature or 37°C to obtain a blank adaptive hydrogel dressing through host-guest complexation and hydrazone bond tandem action, recorded as Gel; its storage modulus is 554.10 Pa.

[0100] Comparative Example 2, Adaptive hydrogel loaded with levofloxacin but not loaded with insulin

[0101] A method for preparing an adaptive drug-loaded hydrogel comprises the following steps:

[0102] S1. Preparation of HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3 solutions: In a clean bench, weigh HA-Cou and HA-ADH respectively into sterile containers, add water or PBS buffer and vortex to dissolve, to prepare HA-Cou and HA-ADH solutions with a concentration of 10 wt% each;

[0103] S2. Preparation of γCD-CHO@LOF: Dissolve γCD-CHO, DSCHO=1.0, 100 mg, 0.07 mmol and LOF, 27.58 mg, 0.07 mmol in 10 mL of deionized water, stir in the dark at room temperature for 48 h, and collect the product after freeze-drying. Then, disperse the crude product in 5 mL of chloroform, collect the insoluble matter by filtration, and obtain γCD-CHO@LOF after vacuum drying.

[0104] 10 mg LOF was dissolved in 10 mL PBS buffer solution to prepare a 1 mg / mL LOF solution, which was then diluted by half with PBS buffer solution to prepare LOF solutions with different concentrations. The absorbance at a wavelength of 294 nm was measured using an enzyme marker to establish a standard curve.

[0105] The absorbance of the γCD-CHO@LOF solution was measured, and the LOF encapsulation efficiency was calculated to be 82.44% and the drug loading efficiency was 19.37% based on the above standard curve.

[0106] S3. Preparation of γCD-CHO@LOF solution: In a clean bench, weigh γCD-CHO@LOF into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a γCD-CHO@LOF solution with a concentration of 10 wt%;

[0107] S4. Prepare an adaptive hydrogel dressing loaded with levofloxacin: mix 100 μL of the HA-Cou solution prepared in step S1 and 63 μL of the γCD-CHO@LOF solution prepared in step S3 to form a host-guest assembly solution; then add 112 μL of the HA-ADH solution prepared in step S1, stir evenly and incubate at room temperature or 37°C to obtain an adaptive hydrogel dressing loaded with levofloxacin through host-guest complexation and hydrazone bond tandem action, denoted as Gel@LOF; its storage modulus is 493.20 Pa, and the concentration of LOF is 4.39 mg / mL.

[0108] Comparative Example 3, Adaptive hydrogel loaded with insulin but not loaded with levofloxacin

[0109] A method for preparing an adaptive drug-loaded hydrogel comprises the following steps:

[0110] S1. Preparation of γCD-CHO, DSCHO=1.0, HA-Cou, DSCou=0.3 and HA-ADH, DSADH=0.3 solutions: In a clean bench, weigh γCD-CHO, HA-Cou and HA-ADH respectively in sterile containers, add water or PBS buffer and vortex to dissolve, to prepare γCD-CHO, HA-Cou and HA-ADH solutions with a concentration of 10 wt% each;

[0111] S2. Prepare insulin solution: In a clean bench, weigh INS into a sterile container, add water or PBS buffer and vortex to dissolve, to prepare a 100 U / mL INS solution;

[0112] S3. Preparation of an insulin-loaded adaptive drug-loaded hydrogel dressing: Mix 100 μL of the HA-Cou solution prepared in step S1 and 63 μL of the γCD-CHO solution to form a host-guest assembly solution; then add 2.8 μL of the INS solution prepared in step S2 and 112 μL of the HA-ADH solution prepared in step S1 in sequence, stir well, and incubate at room temperature or 37°C to obtain an insulin-loaded adaptive hydrogel dressing through host-guest complexation and hydrazone bond tandem action, denoted as Gel@INS; its storage modulus is 537.90 Pa, and the INS concentration is 0.1 U / mL.

[0113] The properties of the hydrogels prepared in the above examples and comparative examples were tested, and the results were as follows:

[0114] Figure 6 The solution-gel transition process of HA-Cou / γCD-CHO@LOF preassembled solution mixed with INS and HA-ADH solutions was recorded.

[0115] Evaluation of tissue adhesion, self-healing and self-adaptability of adaptive drug-loaded hydrogel dressings:

[0116] Tissue adhesion Figure 7 As shown, in order to simulate the actual usage scenario, a certain amount of hydrogel precursor solution dyed with rhodamine B was applied to pig skin, pig liver and pork. After the hydrogel matured, the adhesion of the hydrogel was evaluated by flipping, folding and twisting the above tissues. The test results show that the hydrogel has good tissue adhesion on tissues such as pig skin, pig liver and pork.

[0117] Self-healing Figure 8As shown in the figure, two circular hydrogels stained with rhodamine B and methylene blue were cut into two halves, and half of each semicircular hydrogel was glued together along the cut surface. After half an hour, the healing status of the hydrogel was observed. The test results show that the hydrogel is fully healed and can withstand a certain tensile strength without breaking along the cut surface, indicating that the hydrogel has good self-healing properties.

[0118] Adaptability Fig. 9 As shown in the figure, a cylindrical hydrogel stained with methylene blue was placed on top of the punched pork. After a period of rest, the pork was cut in half and the depth of the hydrogel penetration into the hole was observed. The test results show that the hydrogels with different solid contents all penetrated to a certain depth after standing for 8 hours, and the hydrogels with solid contents of 5 wt% and 10 wt% have penetrated to the bottom of the hole. It can be seen that the hydrogel has good shape adaptability and can adapt to complex wound shapes.

[0119] Biosafety Assessment of Hydrogel Dressings:

[0120] Blood compatibility test: 500 μL of mouse whole blood was taken into an anticoagulant tube, then dispersed into 5 mL of PBS buffer, red blood cells were collected by multiple centrifugation, and resuspended with saline. 500 μL of red blood cell suspension was mixed with 500 μL of hydrogel dressing extract of different concentrations, and 500 μL of red blood cell suspension + 500 μL of saline was used as the negative control group, and 500 μL of red blood cell suspension + 500 μL of ultrapure water was used as the positive control group. They were incubated at 37°C and 120 rpm for 1 h, and the supernatant was centrifuged and its absorbance at a wavelength of 540 nm was measured to calculate the hemolysis rate. Three parallel experiments were performed in each group.

[0121] Cytocompatibility test: The cytotoxic effect of hydrogel dressings on cells was evaluated by MTT experiment and live-dead cell double staining experiment. The specific method is as follows: mouse embryonic fibroblasts, NIH / 3T3, were inoculated in 96- and 24-well plates containing DMEM complete medium and placed in a sterile incubator for 12 h to allow the cells to grow completely attached to the wall. The old culture medium was removed, and fresh culture medium containing the hydrogel dressing extract was added. The cell group cultured in pure culture medium in a 96-well plate was used as a positive control, and pure culture medium was used as a negative control, and was placed in a sterile incubator for 24 and 48 h. 20 μL of MTT solution, 5 mg / mL, was added to each well of the 96-well plate and incubated for another 3.5 h. The MTT solution was removed, 200 μL of DMSO was added to each well, and the absorbance at a wavelength of 570 nm was measured after 15 min to calculate the cell survival rate. 500 μL of buffer containing 0.1% calcein, CalceinAM and 0.1% propidium iodide, PI was added to each well of the 24-well plate. After 30 min, the cells were observed under an inverted fluorescence microscope. Three parallel experiments were performed in each group.

[0122] Evaluation of the antibacterial activity of the hydrogel dressing: Escherichia coli and Staphylococcus aureus were inoculated in Luria-Bertani, LB liquid culture medium, and incubated overnight at 37°C and 120 rpm to activate the bacterial solution. The bacterial solution was diluted to a concentration of 1×10 6 CFU / mL, 900 μL of bacterial solution was mixed with 100 μL of 200 mg / mL hydrogel dressing extract, and the bacterial suspension + PBS buffer solution was used as the positive control group. After incubation for 12 h, the supernatant was taken to measure its absorbance at 600 nm, and the survival rate of bacteria in each group was calculated in turn. Another 100 μL of supernatant was spread on the LB solid culture medium plate, placed in a constant temperature bacterial incubator and inverted for 12 h to observe the growth of the colony.

[0123] Evaluation of the efficacy of hydrogel dressings in healing diabetic infected wounds: C57BL / 6 mice were fed a high-fat diet for one month, and then injected with streptozotocin, 50 mg / mL, for 5 days. When their fasting blood glucose was ≥16.7 mmol / L, a full-thickness skin defect with a diameter of 8 mm and a thickness of 1 mm was made on the back of the mice using a puncher. 20 μL of Staphylococcus aureus, 1×106 bacterial liquid, was dripped onto the wound and infected for 24 h. Different hydrogel dressings were applied to the wound, and the dressings were changed every 2 days. The group with saline dripping was used as the blank control group, and the weight of the mice was recorded to observe the health of the mice. Photos of the mouse wounds were collected on days 0, 3, 7, and 14, and the wound healing rate was calculated to evaluate the healing of the wound.

[0124] Figure 10~Figure 13The biosafety evaluation results of the hydrogel dressing prepared in the present invention are as follows. Fig.10 and Fig.11 The results of the hydrogel dressing hemolysis test showed that the red blood cells treated with the hydrogel dressings in Example 1 and Comparative Examples 1, 2, and 3 all gathered at the bottom of the centrifuge tube, and the supernatant was clear and transparent, with no hemolysis. The hemolysis rates were all lower than 5%, indicating that each group of hydrogel dressings had good blood compatibility. Fig.12 and Fig.13 The results of the hydrogel dressing cytotoxicity experiment showed that the cell survival rates after treatment with different hydrogel dressings were all greater than 80%. The fluorescent photos also showed that the cells basically exhibited green fluorescence, green: living cells; red: dead cells, indicating that each group of hydrogel dressings had good cell compatibility.

[0125] Fig.14 and Fig.15 The results are the inhibition results of the hydrogel dressing prepared in the present invention on Escherichia coli and Staphylococcus aureus. Fig.14 and Fig.15 As shown, the bacterial survival rates of the hydrogel dressings loaded with LOF in Example 1 and Comparative Example 2 were both lower than 20%, and no colonies were generated on the agar plates, while the bacterial survival rates of the hydrogel dressings not loaded with LOF in the blank group, Comparative Example 1 and Comparative Example 3 were all higher than 90%, and the agar plates were full of colonies, indicating that loading LOF gave the hydrogel dressings excellent antibacterial properties.

[0126] Figure 16~Figure 18 The results are as follows: The hydrogel dressing prepared in the present invention promotes the healing of diabetic infected wounds in mice. Fig.16 and Fig.17 The healing process of mouse wounds and the results of mouse wound healing rates show that the hydrogel dressings in Example 1 and Comparative Examples 1, 2, and 3 all have a certain ability to promote wound healing, and the adaptive drug-loaded hydrogel dressing group in Example 1 has the most significant ability to promote wound healing. Fig.18 As shown in the body weight of mice during wound healing, the body weight of mice in each hydrogel dressing treatment group remained basically stable, indicating that the mice were in good health.

[0127] In summary, the adaptive drug-loaded hydrogel dressing prepared by the present invention has better adaptability than traditional hydrogel dressings. It can not only rearrange the gel network, effectively fill and match complex-shaped wounds, and increase the efficiency of drug delivery to the wound, but also achieve efficient loading of antibacterial agents and insulin and long-acting sustained release at the wound, inhibit bacterial infection, promote endothelial cell proliferation, angiogenesis and collagen deposition, thereby significantly accelerating the healing process of diabetic infected wounds.

[0128] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing an adaptive drug-loaded hydrogel, characterized in that: The following steps are involved: Dissolving aldehyde-modified γ-cyclodextrin and an antibacterial agent in water, stirring at room temperature in the dark to obtain a crude product, dispersing the crude product in an organic solvent, filtering to remove the antibacterial agent not included by the aldehyde-modified γ-cyclodextrin to obtain insoluble matter, and drying to obtain an aldehyde-modified γ-cyclodextrin / antibacterial agent inclusion complex; The aqueous solution of the aldehyde-modified γ-cyclodextrin / antimicrobial agent inclusion complex is mixed with the aqueous solution of sodium hyaluronate grafted coumarin, and the γ-cyclodextrin and coumarin produce a host-guest complexation to form a host-guest assembly solution containing the antimicrobial agent. Then, an insulin solution and a sodium hyaluronate grafted adipic acid dihydrazide solution are added, and the mixture is incubated at 20-37° C. The aldehyde group of the host-guest assembly containing the antimicrobial agent undergoes a hydrazone bond coupling reaction with the hydrazide group of the sodium hyaluronate grafted adipic acid dihydrazide to obtain a hydrogel carrier, and the antimicrobial agent and insulin are loaded on the carrier at the same time to obtain an adaptive drug-loaded hydrogel.

2. The preparation method according to claim 1, characterized in that: The concentration of the antibacterial agent in the adaptive drug-loaded hydrogel is 1.10 mg / mL to 6.59 mg / mL.

3. The preparation method according to claim 1, characterized in that: The concentration of the insulin solution in the adaptive drug-loaded hydrogel is 0.1 U / mL~1 U / mL.

4. The preparation method according to claim 1, characterized in that: The antibacterial agent is levofloxacin, curcumin, ciprofloxacin, tetracycline or baicalin.

5. The preparation method according to claim 1, characterized in that: The molar ratio of the gamma-cyclodextrin to coumarin is 1:0.8-1:3, and the molar ratio of the aldehyde group to the hydrazide group is 1:0.5-1:

2.

6. The preparation method according to claim 5, characterized in that: The molar ratio of the gamma-cyclodextrin to coumarin is 1:2, and the molar ratio of the aldehyde group to the hydrazide group is 1:

1.

7. The preparation method according to claim 1, characterized in that: The solid content of the adaptive drug-loaded hydrogel is 5 w / v% to 15 w / v%.

8. The preparation method according to claim 1, characterized in that: Calculated based on the number of aldehyde groups on each aldehyded γ-cyclodextrin molecule, the degree of substitution DSCHO of the aldehyded γ-cyclodextrin is 0.85-1.72; calculated based on the number of moles of coumarin groups per mole of disaccharide unit, the degree of substitution DSCou of sodium hyaluronate grafted coumarin is 0.15-0.60; calculated based on the number of moles of adipic acid dihydrazide groups per mole of disaccharide repeating unit, the degree of substitution DSADH of sodium hyaluronate grafted adipic acid dihydrazide is 0.15-0.

55.

9. An adaptive drug-loaded hydrogel prepared according to the preparation method according to any one of claims 1 to 8.

10. Use of the adaptive drug-loaded hydrogel according to claim 9 in preparing a drug for treating wound infection caused by diabetes.

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