A hydrogel with antibacterial and hemostatic functions and a preparation method and application thereof

Abstract

This invention provides a hydrogel with antibacterial and hemostatic functions. The hydrogel is prepared from the following raw materials: chitosan or chitosan derivatives, lysine acrylamide monomer, methacryloyloxyethyltrimethylammonium chloride monomer, metal ions, and an initiator; the mass ratio of the lysine acrylamide monomer to the methacryloyloxyethyltrimethylammonium chloride monomer is 1–9:1. This hydrogel not only possesses hemostatic function and excellent mechanical properties, but also combats wound infection, shortens the inflammatory cycle, and promotes the repair and regeneration of infected wounds, making it a hydrogel wound dressing with significant application value.

Description

Technical Field

[0001] This invention belongs to the technical field of biomedical materials, and more specifically, relates to a hydrogel with antibacterial and hemostatic functions, its preparation method, and its application. Background Technology

[0002] The skin is the body's first line of defense, preventing the invasion of foreign objects and bacterial infection. When a wound appears, bacteria gather around it, potentially leading to infection if not effectively prevented. The abnormalities caused by wound infection are primarily due to collagen metabolism disorders. In the infected area, neutrophils, after engulfing bacteria, release proteases and oxygen free radicals that damage tissue, causing collagen dissolution to exceed deposition, resulting in delayed wound healing. In the presence of infection, bacteria and inflammatory cells increase oxygen and other nutrient consumption, impair fibroblast metabolism, and increase post-infection exudate, increasing local tension in the wound and even causing it to dehiscence. Wound infection is the most serious disruptive factor in the wound healing process, mainly manifested in the destruction of remaining epithelial tissue, thus delaying healing time. When inflammation leads to suppuration, epithelial growth ceases.

[0003] Ideal wound dressings need to possess certain mechanical properties, self-adhesion, and breathability, as well as good biocompatibility, antibacterial properties, hemostatic properties, and wound-healing properties. For common traumas such as abrasions, lacerations, pressure sores, and chemical injuries, traditional treatments generally involve surgical sutures, medical bandages, and antibiotics. While these methods are widely used, they have limitations. For example, surgical sutures can cause discomfort to patients and leave scars after healing; medical bandages are difficult to use on wounds near joints, and bandages tend to adhere to the wound tissue, potentially causing secondary injury during dressing changes. Therefore, developing novel wound dressings is essential.

[0004] Currently, polymer-based wound dressings include hydrogels, foams, films, and hydrocolloids. Compared with other polymer dressings, hydrogels have significant performance advantages as wound dressings: (1) Hydrogels can achieve cooling and analgesia effects through the evaporation of their own water, and the analgesic effect is sustainable; (2) Hydrogels do not form strong adhesions with human skin tissue, so changing dressings does not cause pain and will not cause secondary damage to the wound; (3) Hydrogels have special pore structures and properties that can meet the needs of different wound environments.

[0005] Methacryloxyethyltrimethylammonium chloride is a cationic monomer that can be polymerized via free radicals. Polymethacryloxyethyltrimethylammonium chloride exhibits antibacterial activity against both Gram-positive and Gram-negative bacteria. However, methacryloxyethyltrimethylammonium chloride itself lacks the ability to accelerate wound healing, and the mechanical properties of hydrogel materials currently developed based on it are also poor. Therefore, providing a hydrogel material with excellent mechanical properties, antibacterial activity, hemostasis, and accelerated repair and regeneration has become an urgent technical problem to be solved. Summary of the Invention

[0006] In view of the above-mentioned existing technical problems, the primary objective of the present invention is to provide a hydrogel with antibacterial and hemostatic functions. The hydrogel wound dressing not only has antibacterial and hemostatic functions and excellent mechanical properties, but also can resist infection of the wound, shorten the inflammation cycle, and promote the repair and regeneration of infected wounds.

[0007] A second objective of this invention is to provide a method for preparing the above-mentioned hydrogel with antibacterial and hemostatic functions.

[0008] The third objective of this invention is to provide a hydrogel wound dressing with antibacterial and hemostatic functions.

[0009] A fourth objective of this invention is to provide the application of the above-mentioned hydrogel or hydrogel wound dressing with antibacterial and hemostatic functions in the preparation of wound repair materials.

[0010] To achieve the above objectives, the present invention is implemented through the following technical solution:

[0011] A hydrogel with antibacterial and hemostatic functions, the hydrogel being prepared from the following raw materials: chitosan or chitosan derivatives, lysine acrylamide monomer, methacryloyloxyethyltrimethylammonium chloride monomer, metal ions, and an initiator; wherein the mass ratio of lysine acrylamide monomer to methacryloyloxyethyltrimethylammonium chloride monomer is 1 to 9:1.

[0012] This invention provides a hydrogel with antibacterial and hemostatic functions. Under the action of chitosan or chitosan derivatives and an initiator, a specific ratio of lysine acrylamide monomers and methacryloyloxyethyltrimethylammonium chloride monomers can undergo free radical polymerization to generate linear poly(lysine acrylamide-copolymer-methacryloyloxyethyltrimethylammonium chloride) macromolecular chains. Subsequently, the carboxyl groups of the side chains of the macromolecular chains form metal ion-carboxylate coordination bonds with metal ions, thereby forming a cross-linked network structure. Simultaneously, the functional groups of the chitosan derivatives can also reversibly interact with the poly(lysine acrylamide-copolymer-methacryloyloxyethyltrimethylammonium chloride) macromolecular chains through hydrogen bonds, ionic bonds, and other mechanisms, thus forming a cross-linked network structure. This invention eliminates the need for chemical cross-linking agents, relying on the formation of metal ion-carboxylate coordination bonds by metal ions and physical interactions (hydrogen bonds, ionic interactions) between macromolecular chains to form a cross-linked network structure, thereby preparing a hydrogel.

[0013] The hydrogel provided by this invention exhibits excellent bactericidal and hemostatic effects, good swelling properties, good biocompatibility, and promotes skin tissue healing, demonstrating significant practical application value. When the hydrogel comes into contact with bacteria such as Staphylococcus aureus and Escherichia coli, the cations on the hydrogel surface combine with the anions on the bacterial surface, causing the bacterial membrane to rupture. This achieves the inactivation of Gram-positive and Gram-negative bacteria, including Staphylococcus aureus and Escherichia coli, while also preventing the development of bacterial resistance. Furthermore, the cations on the hydrogel surface are bound to the material and are not easily released, thus extending the antibacterial period of the hydrogel. When used as a wound dressing, the hydrogel's excellent swelling properties allow blood to be adsorbed within its three-dimensional network structure, causing clotting factors to accumulate at the wound site. Simultaneously, the positively charged functional groups in the hydrogel can combine with negatively charged red blood cells to form a blood clot, achieving hemostasis. The clotting process is a physical process without any adverse reactions. Simultaneously, the hydrogel absorbs tissue fluid exudated from the wound, stimulates the proliferation of fibroblasts in the wound, promotes collagen deposition, regulates macrophage polarization towards the anti-inflammatory and healing-promoting M2 type, and stimulates capillary regeneration, thereby promoting wound tissue regeneration. Therefore, the hydrogel developed in this invention has significant application value in the field of wound repair.

[0014] Preferably, the mass ratio of the lysine acrylamide monomer to the methacryloyloxyethyltrimethylammonium chloride monomer is 1 to 3:1; most preferably, the mass ratio is 1:1.

[0015] Preferably, the mass ratio of the copolymer formed by the chitosan or chitosan derivative with lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer is 1:5 to 20. More preferably, the mass ratio is 1:8 to 15; most preferably, the mass ratio is 1:10.

[0016] Preferably, the molar mass ratio of the copolymer formed by the metal ion and lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer is (0.001-0.032):(1.5-6) ​​mmol / g; more preferably, the molar mass ratio is (0.005-0.015):(2-4) mmol / g; most preferably, the molar mass ratio is 0.008:3 mmol / g.

[0017] Preferably, the metal ion is selected from one or more of iron ions, aluminum ions, and zinc ions. More preferably, the metal ion is an iron ion. More specifically, the metal ion can participate in the synthesis of hydrogels in the form of various metal salts or metal salt solutions, such as nitrates, hydrochlorides, etc.

[0018] Preferably, the initiator is selected from one or more of ammonium persulfate, potassium persulfate, and sodium persulfate. More preferably, the initiator is ammonium persulfate.

[0019] Preferably, the concentration range of the initiator is 1-5% (w / v). More preferably, the concentration range of the initiator is 1-3% (w / v); most preferably, the concentration of the initiator is 2% (w / v).

[0020] Preferably, the chitosan derivative is selected from one or both of quaternized chitosan and carboxymethyl chitosan.

[0021] Furthermore, the present invention also claims protection for a method for preparing a hydrogel with antibacterial and hemostatic functions, wherein lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer are mixed with chitosan or chitosan derivative-acetic acid solution, and then metal ions and an initiator are added and mixed to obtain the hydrogel.

[0022] Furthermore, as a specific embodiment of the present invention, a method for preparing a hydrogel with antibacterial and hemostatic functions includes the following steps: dissolving chitosan or chitosan derivatives in an acetic acid solution with a concentration of 1-3% (w / v) to obtain a solution A (i.e., chitosan or chitosan derivative-acetic acid solution) with a chitosan concentration of 0-4%; adding lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer to solution A to obtain solution B; adding a metal ion solution and an ammonium persulfate solution with a concentration of 1-5% (w / v) to solution B, mixing thoroughly, and then initiating a free radical polymerization reaction at 60-70°C to obtain a hydrogel with antibacterial and hemostatic functions.

[0023] More preferably, the concentration of chitosan or chitosan derivative in the chitosan or chitosan derivative-acetic acid solution is 3-4%.

[0024] More preferably, an ammonium persulfate solution is added, and after thorough mixing, a free radical polymerization reaction is initiated at 60°C for 2–10 hours.

[0025] Furthermore, the present invention also claims protection for a hydrogel wound dressing with antibacterial and hemostatic functions, which is obtained by soaking the above-mentioned hydrogel with antibacterial and hemostatic functions in a sodium chloride solution.

[0026] More preferably, the soaking time is 1 to 50 minutes. More preferably, the soaking time is 5 to 20 minutes.

[0027] Preferably, the sodium chloride solution is a saturated sodium chloride solution.

[0028] Furthermore, the present invention also claims protection for the use of the above-mentioned hydrogel with antibacterial and hemostatic functions or the above-mentioned hydrogel wound dressing with antibacterial and hemostatic functions in the preparation of wound repair materials.

[0029] Compared with the prior art, the present invention has the following beneficial effects: the hydrogel provided by the present invention has excellent bactericidal and hemostatic effects, good swelling properties, good biocompatibility, and promotes skin tissue healing. It also has a longer antibacterial cycle. The hydrogel developed by the present invention has important application value in the field of wound repair. Attached Figure Description

[0030] Figure 1 The hydrogel wound dressing prepared in Example 6 is shown before and after formation; wherein Figure 1 a represents the state before hydrogel formation; Figure 1 b represents the state after hydrogel formation; Figure 1 c represents the hydrogel before being soaked in saline solution; Figure 1 d represents the hydrogel after being soaked in salt water.

[0031] Figure 2 The infrared spectrum of the hydrogel wound dressing prepared in Example 6.

[0032] Figure 3 Stress-strain curves of hydrogels or hydrogel wound dressings prepared for examples and comparative examples.

[0033] Figure 4 Optical and fluorescence images of L929 cells treated with hydrogel or hydrogel wound dressing extract at a concentration of 500 μg / mL.

[0034] Figure 5 The effect of a 100 μg / mL hydrogel or hydrogel wound dressing extract on cell viability.

[0035] Figure 6The effect of a hydrogel or hydrogel wound dressing extract at a concentration of 500 μg / mL on cell viability.

[0036] Figure 7 The effect of a 1000 μg / mL hydrogel or hydrogel wound dressing extract on cell viability.

[0037] Figure 8 The hemolysis rate of hydrogels or hydrogel wound dressings.

[0038] Figure 9 The image shows the inhibitory effect of hydrogels or hydrogel wound dressings on Bacillus subtilis.

[0039] Figure 10 This image shows the inhibitory effect of hydrogels or hydrogel wound dressings on Staphylococcus aureus.

[0040] Figure 11 This image shows the inhibitory effect of hydrogels or hydrogel wound dressings on Escherichia coli.

[0041] Figure 12 The plate colony counts are those of Bacillus subtilis, Staphylococcus aureus, and Escherichia coli cultures after being treated with hydrogel or hydrogel wound dressing.

[0042] Figure 13 To characterize the blood absorption capacity of hydrogels or hydrogel wound dressings, among which... Figure 13 a represents the time before blood is added; Figure 13 b represents the result after blood was added; Figure 13 c represents the result after adding deionized water.

[0043] Figure 14 BCI test results for hydrogels or hydrogel wound dressings.

[0044] Figure 15 APTT test results for hydrogels or hydrogel wound dressings.

[0045] Figure 16 The results of PT experiments for hydrogels or hydrogel wound dressings.

[0046] Figure 17 Optical photographs of a tail-cutting hemostasis experiment using hydrogel wound dressings.

[0047] Figure 18 Blood loss and hemostasis time were measured during a tail amputation hemostasis experiment on hydrogel wound dressings. Detailed Implementation

[0048] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0049] Example 1: Preparation of hydrogels with antibacterial and hemostatic functions

[0050] (1) Dissolve 0.3g of chitosan in 10mL of 2% (w / v) acetic acid solution to obtain solution A;

[0051] (2) Lysine acrylamide and methacryloyloxyethyltrimethylammonium chloride were added to solution A, with a total mass of 3.0 g and a mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride of 9:1, to obtain solution B;

[0052] (3) Add 0.016 mmol of iron ion solution (iron ion solution prepared by iron nitrate nonahydrate) and 1 mL of 2% (w / v) ammonium persulfate solution to solution B, mix thoroughly, and heat at 60°C to initiate free radical polymerization reaction for 6 h to obtain hydrogel with antibacterial and hemostatic functions.

[0053] Example 2

[0054] The difference between this embodiment and embodiment 1 is that in step (2), the mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride is 7:3.

[0055] Example 3

[0056] The difference between this embodiment and embodiment 1 is that in step (2), the mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride is 1:1.

[0057] Example 4

[0058] The difference between this embodiment and embodiment 1 is that: in step (2), the mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride is 1:1; in step (3), the hydrogel wound dressing with antibacterial and hemostatic functions is obtained by soaking in saturated sodium chloride solution for 10 minutes.

[0059] Example 5

[0060] The difference between this embodiment and embodiment 1 is that: in step (2), the mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride is 1:1; in step (3), 0.004 mmol of iron ion solution is added to solution B; after the reaction is completed, it is soaked in saturated sodium chloride solution for 10 min to obtain a hydrogel wound dressing with antibacterial and hemostatic functions.

[0061] Example 6

[0062] The difference between this embodiment and embodiment 1 is that: in step (2), the mass ratio of lysine acrylamide to methacryloyloxyethyltrimethylammonium chloride is 1:1; in step (3), 0.008 mmol of iron ion solution is added to solution B; after the reaction is completed, it is soaked in saturated sodium chloride solution for 10 min to obtain a hydrogel wound dressing with antibacterial and hemostatic functions.

[0063] Example 7

[0064] The difference between this embodiment and embodiment 1 is that: in step (2), the total mass of lysine acrylamide and methacryloyloxyethyltrimethylammonium chloride is 1.5g; in step (3), an aluminum ion solution is used, and the aluminum ion solution is 0.032mmol.

[0065] Example 8

[0066] The difference between this embodiment and embodiment 1 is that: in step (2), the total mass of lysine acrylamide and methacryloyloxyethyltrimethylammonium chloride is 6g; in step (3), a zinc ion solution is used, and the zinc ion solution is 0.001mmol.

[0067] Comparative Example 1

[0068] The difference between this comparative example and Example 3 is that: in step (1), the chitosan content is 0 g; in step (2), 0 mmol of iron ion solution is added to solution B.

[0069] Comparative Example 2

[0070] The difference between this comparative example and Example 3 is that in step (2), 0 mmol of iron ion solution is added to solution B.

[0071] Comparative Example 3

[0072] The difference between this comparative example and Example 3 is that in step (1), the amount of chitosan is 0g.

[0073] Comparative Example 4

[0074] The difference between this comparative example and Example 1 is that in step (2), only lysine acrylamide is added, and methacryloyloxyethyltrimethylammonium chloride is not added.

[0075] Test Example 1: Characterization of hydrogel wound dressings with antibacterial and hemostatic functions

[0076] like Figure 1 The images show the hydrogel wound dressing prepared in Example 6 of this invention before and after formation; wherein, Figure 1 a represents the state before hydrogel formation; Figure 1 b represents the state after hydrogel formation; Figure 1 c represents the hydrogel before being soaked in saline solution; Figure 1 d represents the hydrogel after being soaked in saline solution; Figure 2 Infrared spectrum of the hydrogel wound dressing prepared in Example 6. Figure 1 and Figure 2 It can be seen that the hydrogel and hydrogel wound dressing in Example 6 of the present invention were successfully prepared.

[0077] Test Example 2: Mechanical Properties and Moisture Content Testing of Hydrogels or Hydrogel Wound Dressings with Antibacterial and Hemostatic Functions

[0078] The mechanical property testing methods for the hydrogels or hydrogel wound dressings prepared in the above embodiments and comparative examples are as follows: Tensile tests were performed on the samples using a universal testing machine at a tensile rate of 100 mm / min and a room temperature. Five samples were tested for each sample, and the average value was used to calculate the tensile strength and elongation at break. The moisture content testing method is as follows: The freshly prepared hydrogel was weighed (M1), and then dried in a 60℃ oven until its mass became constant. The mass M2 after drying was measured, and the moisture content was calculated using the formula: Q = (M1 - M2) / M1 × 100%. The test results are shown in Table 1 below. Figure 3 As shown, Figure 3 Stress-strain curves of hydrogels or hydrogel wound dressings prepared for examples and comparative examples.

[0079] Table 1

[0080]

[0081]

[0082] As shown in Table 1, the hydrogels prepared in Comparative Examples 1, 3, and 4 showed no relevant mechanical properties. Regarding mechanical properties, the tensile strength and elongation at break of Examples 5 and 6 were significantly better than those of the other examples.

[0083] Test Example 3: Biocompatibility of hydrogels with antibacterial and hemostatic functions

[0084] The cytotoxic effects of the hydrogel wound dressings prepared in Comparative Examples 1-2 and Examples 1-6 were evaluated using a CCK-8 assay kit and a live-dead staining kit. The test methods were as follows: (1) CCK-8 assay: Mouse fibroblast L929 cells were seeded in 96-well plates, and the material extracts (concentrations of 100 μg / mL, 500 μg / mL, and 1000 μg / mL, respectively) were co-cultured with the cells for 24 h, 48 h, and 72 h. Cell proliferation was detected by the CCK-8 assay. (2) Hemolysis rate: A certain amount of hydrogel samples prepared in Comparative Examples 1-2 and Examples 1-6 were added to 2.5% (v / v) red blood cell suspension. 0.05 mL of red blood cells were added to 2 mL of deionized water and 2 mL of PBS solution, respectively. The samples were cultured at 37°C and 100 rpm for 1 h, and the absorbance was measured to calculate the hemolysis rate. Each group was tested in triplicate.

[0085] Figure 4 Optical and fluorescence images of L929 cells treated with a hydrogel or hydrogel wound dressing extract at a concentration of 500 μg / mL. Figures 5-7 The effects of hydrogel or hydrogel wound dressing extracts at concentrations of 100 μg / mL, 500 μg / mL, and 1000 μg / mL on cell viability were investigated. Figures 4-7 It is evident that the hydrogels or hydrogel wound dressings prepared in the examples and comparative examples did not exhibit significant cytotoxicity.

[0086] Figure 8 For hemolysis rate of hydrogels or hydrogel wound dressings, such as Figure 8 As shown, the hemocompatibility of hydrogel wound dressings was evaluated by hemolysis test. The hemolysis rate of hydrogel or hydrogel wound dressing was less than 5%, and it did not cause a hemolytic reaction, thus exhibiting excellent blood compatibility.

[0087] Test Example 4: Antibacterial Experiment of Hydrogels with Antibacterial and Hemostatic Functions

[0088] The antibacterial properties of the hydrogel wound dressings from Comparative Examples 1-2 and Examples 1-6 were tested. The test method was as follows: A certain amount of sterilized hydrogel samples from Comparative Examples 1-2 and Examples 1-6 were added to 5 mL of bacterial solution. A blank control group was set up. The samples were incubated at 100 rpm and 37℃ for a period of time. The OD values ​​of the bacterial solution were measured at 2 h, 6 h, 12 h, and 24 h to obtain the bacterial growth curves. 100 μL of the bacterial solution treated with hydrogel for 24 h was further incubated on LB solid medium for 12 h. Images were taken, and the number of colonies was counted using the plate count method to calculate the inhibition rate. Each group was tested in triplicate. The test and calculation results are shown in Table 2. The calculated antibacterial rate reached 99%.

[0089] Table 2

[0090]

[0091] Figures 9-11 The images show the inhibitory effects of hydrogels or hydrogel wound dressings on Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, respectively. Figure 9 In the control group, the OD values ​​of the bacterial solutions treated with hydrogel or hydrogel dressings were generally lower. Figure 10 and Figure 11 In the study, only a portion of the bacterial solutions treated with hydrogel or hydrogel dressings showed lower OD values ​​than the control group. This indicates that the antibacterial ability of hydrogels or hydrogel dressings is related to their composition and preparation process. Figure 12 This image shows the plate colony counts after culturing Bacillus subtilis, Staphylococcus aureus, and Escherichia coli cultures treated with hydrogel or hydrogel wound dressings. Figure 12 As shown in Table 2, the antibacterial ability of hydrogels or hydrogel dressings is related to their composition and preparation process. Hydrogel dressings prepared with better composition and preparation process have good inhibitory effects on Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. In addition, the data above also show that after hydrogels are soaked in sodium chloride solution, the polymer chains are induced to become more entangled and cross-linked. Under the same volume, hydrogels soaked in sodium chloride solution contain more amino, quaternary ammonium groups and cationic groups, which is beneficial to improving their antibacterial properties.

[0092] Test Example 5: Hemostasis Experiment of Hydrogels with Antibacterial and Hemostatic Functions

[0093] The hemostatic properties of the hydrogel wound dressings prepared in Comparative Examples 1-2 and Examples 1-6 were evaluated using BCI, APTT, PT tests, and tail amputation hemostasis tests. The test methods are as follows:

[0094] (1) BCI: 0.1M calcium chloride solution was added to anticoagulated rabbit whole blood at a ratio of 1:10 (v / v) to obtain activated blood. Then, 20 μL of activated blood was quickly dropped onto the surface of hydrogel or hydrogel wound dressing samples prepared in Comparative Examples 1-2 and Examples 1-6 (or blank control). After 5 min, 25 mL of deionized water was slowly added. After 10 min, the image was taken. Finally, the hemoglobin solution released by the uncoagulated red blood cells was collected, and its OD value at 540 nm was measured to calculate BCI. Each group was tested in triplicate.

[0095] (2) APTT: Take 0.1 ml of the plasma to be tested, add 5 mg of the hydrogel or hydrogel wound dressing sample prepared in Comparative Examples 1-2 and Examples 1-6, and 0.1 ml of APTT reagent preheated to 37°C, and incubate at 37°C for 5 min. Add 0.1 ml of 0.025 mol / L CaCl2 solution preheated to 37°C, and record the APTT coagulation time. Perform three parallel experiments for each group.

[0096] (3) PT: Take 0.1 ml of the plasma to be tested, incubate at 37°C for 3 min, add 5 mg of hydrogel or hydrogel wound dressing samples prepared in Comparative Examples 1-2 and Examples 1-6, and 0.2 ml of PT reagent preheated at 37°C. Record the coagulation time, which is the PT value. Each group is tested in 3 parallel experiments.

[0097] (4) Tail amputation hemostasis experiment: Anesthetized mice were fixed on a sterilized foam board, and the tail tip (50% of the total tail length) was cut off, allowing blood to flow onto pre-weighed filter paper. A 6mm diameter sample of the hydrogel wound dressing from Example 6 was then applied to the bleeding site with gentle pressure, and a photograph was taken every 60 seconds. The untreated group served as the control group. Time was recorded from the application of the dressing to the amputated tail until bleeding stopped. Blood loss was calculated, and each group underwent three parallel experiments.

[0098] Figure 13 An in vitro coagulation effect diagram of hydrogel or hydrogel wound dressing. Figures 14-16 This refers to the results of in vitro coagulation performance tests on hydrogels or hydrogel wound dressings. Figures 13-16 It is known that blood coagulation is better in the presence of hydrogel or hydrogel wound dressing than in the natural state.

[0099] Figure 17 This image shows the hemostatic effect of a hydrogel wound dressing after tail cutting. Figure 18 The hemostatic properties of the hydrogel wound dressing. (By...) Figures 17-18 It is known that hydrogel wound dressings can reduce blood loss and shorten the hemostasis time, thus demonstrating that hydrogel wound dressings have hemostatic properties.

[0100] The foregoing examples are merely illustrative, used to explain some features of the method described in this invention. The appended claims are intended to claim the broadest possible scope, and the embodiments presented herein are demonstrated by the applicant's actual experimental results. Therefore, the applicant intends that the appended claims are not limited by the selection of examples illustrating the features of the invention. Some numerical ranges used in the claims also include sub-ranges within them, and variations within these ranges should also be interpreted as being covered by the appended claims where possible.

Claims

1. A hydrogel with antibacterial and hemostatic functions, characterized in that, The hydrogel is prepared from the following raw materials: chitosan or chitosan derivative, lysine acrylamide monomer, methacryloyloxyethyltrimethylammonium chloride monomer, metal ions, and initiator; the mass ratio of lysine acrylamide monomer to methacryloyloxyethyltrimethylammonium chloride monomer is 1 to 9:

1. The metal ion is selected from one or more of iron ions, aluminum ions, and zinc ions.

2. The hydrogel according to claim 1, characterized in that, The mass ratio of the lysine acrylamide monomer to the methacryloyloxyethyltrimethylammonium chloride monomer is 1 to 3:

1.

3. The hydrogel according to claim 2, characterized in that, The mass ratio of the lysine acrylamide monomer to the methacryloyloxyethyltrimethylammonium chloride monomer is 1:

1.

4. The hydrogel according to claim 1, characterized in that, The mass ratio of the copolymer formed by the chitosan or chitosan derivative with lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer is 1:5 to 20.

5. The hydrogel according to claim 1, characterized in that, The molar mass ratio of the copolymer formed by the metal ion with lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer is (0.001-0.032):(1.5-6).

6. The hydrogel according to claim 5, characterized in that, The molar mass ratio of the copolymer formed by the metal ion with lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer is (0.005-0.015):(2-4).

7. The hydrogel according to claim 6, characterized in that, The molar mass ratio of the copolymer formed by the metal ions, lysine acrylamide monomer, and methacryloyloxyethyltrimethylammonium chloride monomer is 0.008:

3.

8. The hydrogel according to any one of claims 1 to 7, characterized in that, The initiator is selected from one or more of ammonium persulfate, potassium persulfate, and sodium persulfate.

9. The hydrogel according to any one of claims 1 to 7, characterized in that, The concentration range of the initiator is 1–5% (w / v).

10. A method for preparing the hydrogel with antibacterial and hemostatic functions according to any one of claims 1 to 9, characterized in that, Lysine acrylamide monomer and methacryloyloxyethyltrimethylammonium chloride monomer are mixed with chitosan or chitosan derivative-acetic acid solution, and then metal ions and initiators are added and mixed to obtain the hydrogel with antibacterial and hemostatic functions.

11. A hydrogel wound dressing with antibacterial and hemostatic functions, characterized in that, The hydrogel with antibacterial and hemostatic functions described in any one of claims 1 to 9 is soaked in sodium chloride solution to obtain the hydrogel wound dressing with antibacterial and hemostatic functions.

12. The application of the hydrogel with antibacterial and hemostatic functions as described in any one of claims 1 to 9, or the hydrogel wound dressing with antibacterial and hemostatic functions as described in claim 11, in the preparation of wound repair materials.

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