Injectable hydrogel for rapid hemostasis and antibiosis of oral cavity as well as preparation method and application of injectable hydrogel

Abstract

The invention relates to injectable hydrogel for rapid hemostasis and antibiosis of an oral cavity as well as a preparation method and application of the injectable hydrogel, and the injectable hydrogel comprises the following components in percentage by weight: 5-15% of aminated chondroitin sulfate, 5-15% of oxidized dextran, 1-3% of platelet-rich fibrin and 0.5-1% of quercetin modified zeolite imidazole acid framework nanoparticles. According to the invention, Chs-ADH and ODex are adopted as matrix materials, PRF and ZIF-8 (at) QCT are introduced, the hydrogel is endowed with antibacterial, anti-oxidation and immunoregulation functions, and by optimizing the proportion of the components, the obtained hydrogel has good injectability, stability and degradation resistance, so that the hydrogel can still maintain an effective blood coagulation function in a high-humidity and high-enzymatic-activity oral environment.

Description

Technical Field

[0001] The invention relates to the field of biomaterials, and in particular to an injectable hydrogel for rapid oral hemostasis and antibacterial properties, and a preparation method and application thereof. Background Art

[0002] Oral hemostasis refers to the use of rapid and effective means to control bleeding in the oral cavity. Common causes of bleeding include dental surgery, trauma, periodontal disease, and oral ulcers. At present, hemostatic hydrogels are widely studied and used in clinical practice. Their main sources of materials include natural polymer materials and synthetic polymers. Natural polymer materials such as gelatin, chitosan, and alginate have attracted attention due to their good biocompatibility. Synthetic polymer materials such as polylactic acid (PLA) and polyvinyl alcohol (PVA) are widely used due to their controllable degradability and good physical properties. These materials usually adsorb blood and promote coagulation by forming a three-dimensional network structure.

[0003] However, due to the complexity of the oral environment and the increasing frequency of trauma, traditional single hemostatic materials and methods have certain defects and shortcomings.

[0004] (1) Poor adaptability: Although gelatin is derived from animal collagen and has good coagulation and biodegradability, it is easily degraded due to the high humidity and strong enzyme activity in the oral cavity, making it difficult to ensure coagulation function, resulting in a short-lasting hemostatic effect.

[0005] (2) Insufficient injection operability: Some hydrogels are prone to clogging and uneven distribution during injection, especially in hidden areas such as the alveolar socket, which affects the hemostatic effect. For example, chitosan, due to its high viscosity, makes the material not convenient during injection.

[0006] (3) Poor effectiveness of hemostatic effect: Many hydrogel products have a long hemostatic time, low recurrence rate of bleeding, and lack of long-term support.

[0007] (4) Poor biocompatibility: Although synthetic polymer materials have controllable degradability and good physical properties, they have poor compatibility and may cause local inflammation, limiting their safety in oral applications.

[0008] In short, the existing hemostatic materials are not multifunctional enough, lacking anti-oxidation, antibacterial, anti-inflammatory and healing-promoting functions, and are difficult to cope with the complex oral environment. Therefore, it is urgent to improve the existing materials and provide a hemostatic material with composite functions. Summary of the invention

[0009] In view of the above-mentioned deficiencies in the prior art, the present invention proposes an injectable hydrogel for rapid oral hemostasis and antibacterial properties, which can rapidly stop bleeding, resist bacteria, promote healing, and has good biocompatibility.

[0010] The specific technical scheme of the present invention is as follows:

[0011] The first aspect of the present invention is an injectable hydrogel for rapid oral hemostasis and antibacterial, comprising the following components in weight percentage: 5-15% amino chondroitin sulfate (Chs-ADH), 5-15% oxidized dextran (ODex), 1-3% platelet-rich fibrin (PRF), and 0.5-1% quercetin-modified zeolite imidazolate framework nanoparticles (ZIF-8@QCT).

[0012] Among them, aminated chondroitin sulfate (Chs-ADH) and oxidized dextran (ODex) are matrix materials with good biocompatibility, anti-degradation and cell adhesion ability, cross-linking to form a hydrogel-based three-dimensional network, platelet-rich fibrin (PRF) can release platelets and production factors, has hemostatic and tissue regeneration effects, and quercetin-modified zeolite imidazolate framework nanoparticles (ZIF-8@QCT) have pH response function and can intelligently release Zn 2+ , providing antibacterial effects and promoting angiogenesis in the oral environment, quercetin has enhanced antibacterial, anti-inflammatory and tissue repair functions; and ZIF-8@QCT can enhance the stability and anti-degradation properties of the hydrogel. The synergistic cooperation of the components can improve the injectability of the hydrogel, the stability of the oral environment, etc., improve the hemostatic effect, anti-inflammatory and antibacterial, promote blood vessel and tissue regeneration, preferably, containing the following weight percentages of raw materials, 10-15% amino chondroitin sulfate, 10-15% oxidized dextran, 1-3% platelet-rich fibrin, 0.5-1% quercetin-modified zeolite imidazole framework nanoparticles.

[0013] Preferably, according to the weight ratio, the amino chondroitin sulfate: oxidized dextran = 1-1.5:1, which has better injectability and better anti-degradation properties.

[0014] Preferably, the aminated chondroitin sulfate is prepared by the following method, including: dissolving sodium chondroitin sulfate and adipic acid diacyl in deionized water to obtain solution I; dissolving 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1-hydroxybenzotriazole hydrate in a dimethyl sulfoxide / water mixture to obtain solution II, then adding solution II dropwise to solution I, adjusting pH to 5.0, reacting at room temperature, and ending the reaction, dripping ethanol for precipitation, dissolving the precipitate in water, dialyzing the solution in pure water, the dialysis molecular weight cutoff is 500Da, and then freeze-drying to obtain aminated chondroitin sulfate. Through modification, a more stable three-dimensional network structure can be formed, with good biocompatibility, anti-degradation and cell adhesion ability, adapting to the high humidity and high enzyme environment of the oral cavity, and providing long-term hemostatic support.

[0015] Further preferably, the dimethyl sulfoxide / water mixture is a mixture of dimethyl sulfoxide and water in a volume ratio of 1:1; sodium chondroitin sulfate: diacyl adipic acid: deionized water: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloric acid: 1-hydroxybenzotriazole hydrate: dimethyl sulfoxide / water mixture = 0.5g: 10g: 100mL: 0.8g: 0.7g: 10mL.

[0016] Preferably, the quercetin-modified zeolite imidazolate framework nanoparticles are prepared by the following method, including: dissolving a soluble zinc salt in deionized water at room temperature to obtain a zinc solution; dissolving 2-methylimidazole and quercetin in methanol to obtain a quercetin solution; then mixing the zinc solution with the quercetin solution, stirring the reaction until the powder is completely dissolved; finally, centrifuging the obtained yellow product, washing it with methanol several times, and vacuum drying it at 30-40°C to obtain the quercetin-modified zeolite imidazolate framework nanoparticles ZIF-8@QCT. Combining the zeolite imidazolate framework with quercetin, on the one hand, the framework structure can realize Zn 2+ The sustained release of quercetin can further enhance the antibacterial, anti-inflammatory and tissue repair functions.

[0017] More preferably, the zinc solution contains Zn 2+ The concentration is 0.08-0.15 mol / L. In the quercetin solution, the concentration of 2-methylimidazole is 0.40-0.5 mol / L, the concentration of quercetin is 1.325 mmol / L, and the volume ratio of the zinc solution to the quercetin solution is 1:2.

[0018] The oxidized dextran (ODex) used in the injectable hydrogel of the present invention is obtained by oxidation of dextran and sodium periodate. In a preferred embodiment, the oxidized dextran is obtained by reacting dextran and sodium periodate at room temperature in the dark, dialyzing through a 8000Da dialysis bag, and freeze-drying. Preferably, the weight ratio of dextran to sodium periodate is 1:1.

[0019] Platelet-rich fibrin can be a commercially available freeze-dried powder, which is usually obtained by centrifuging whole blood, then taking out the middle PRF gel layer, freeze-drying and grinding it to obtain freeze-dried platelet-rich fibrin powder.

[0020] In another aspect of the present invention, a method for preparing the injectable hydrogel is provided, comprising the steps of: dissolving the aminated chondroitin sulfate in a first solvent to obtain an aminated chondroitin sulfate hydrogel solution, and then adding platelet-rich fibrin lyophilized powder to the aminated chondroitin sulfate hydrogel solution in an ice bath environment to obtain a first hydrogel solution;

[0021] adding oxidized dextran to a second solvent to obtain an oxidized dextran solution, and then adding quercetin-modified zeolite imidazolate framework nanoparticles to the oxidized dextran solution for dispersion to obtain a second hydrogel solution;

[0022] The first hydrogel solution and the second hydrogel solution are mixed, and the pH is adjusted to 7-8 to obtain the injectable hydrogel.

[0023] The first solvent and the second solvent are common solvents that can dissolve oxidized dextran or amino chondroitin sulfate to form a hydrogel. Preferably, the first solvent and the second solvent are any one of ultrapure water, PBS buffer, and physiological saline.

[0024] In one aspect, the present invention further provides a wound dressing, comprising the injectable hydrogel or the injectable hydrogel prepared by the preparation method, which is particularly suitable for oral hemostatic dressing.

[0025] The solution of the present invention has the following beneficial effects:

[0026] (1) Improving environmental adaptability: The present invention uses aminated chondroitin sulfate (Chs-ADH) and ODex as matrix materials, and introduces zeolite imidazole acid framework nanoparticles to enhance the stability and anti-degradation properties of the hydrogel, so that it can still maintain effective coagulation function in the oral environment of high humidity and high enzyme activity, thus solving the problem of poor adaptability of existing hydrogels.

[0027] (2) Improve injection operability and distribution uniformity: Through the appropriate ratio of aminated chondroitin sulfate (Chs-ADH) and ODex, dynamic chemical bonds are cross-linked to form a three-dimensional network structure, which enhances the fluidity of the hydrogel, making it less likely to be blocked during injection and able to be evenly distributed in the complex oral structure, ensuring efficient hemostatic coverage.

[0028] (3) Enhance the sustainability of hemostatic effect: The preparation method of the present invention loads platelet-rich fibrin (PRF) into the Chs-ADH matrix. PRF continuously releases platelets and growth factors, provides long-term hemostatic support and promotes tissue regeneration, and is suitable for sustained hemostasis needs after oral surgery.

[0029] (4) Improving biological functional diversity: By introducing quercetin-modified zeolite imidazolate framework (ZIF-8@QCT), QCT can be gradually released, giving the hydrogel antibacterial, antioxidant and immunomodulatory functions and improving its healing ability. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is an image of the injectable hydrogel used for oral hemostasis in Example 7 of the present invention;

[0031] Figure 2A schematic diagram of a PRF extraction position according to an embodiment of the present invention. DETAILED DESCRIPTION

[0032] The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0033] The aminated chondroitin sulfate, PRF freeze-dried powder, quercetin-modified zeolite imidazolate framework nanoparticles, and oxidized dextran used in the following examples were prepared by the following method.

[0034] (1) Preparation of aminated chondroitin sulfate Chs-ADH:

[0035] Weigh 0.5g of sodium chondroitin sulfate (Chs) and 10g of diacyl adipic acid (ADH), dissolve them in 100mL of deionized water and stir evenly. Weigh 0.8g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 0.7g of 1-hydroxybenzotriazole hydrate (HOBT), dissolve them in a mixture of dimethyl sulfoxide / water (v / v=1:1, 5ml each), add the mixture dropwise to the above solution, adjust the pH to 5.0 with dilute hydrochloric acid, and react at room temperature for 1 day. After the reaction is completed, add ethanol dropwise for precipitation, dissolve the precipitate in water, dialyze the solution in pure water (molecular weight cutoff: 500Da) for 3-5 days, and freeze-dry for 48h to obtain amino chondroitin sulfate, recorded as Chs-ADH.

[0036] (2) PRF preparation

[0037] 8 mL of whole blood was collected and centrifuged (3000 rpm, 10 min). After centrifugation, the blood components were separated into layers, such as Figure 2 As shown, the upper layer: cell-free plasma (PPP) - mainly platelets and fibrin; the middle layer: platelet-rich fibrin (PRF); the lower layer: red blood cells. After centrifugation, use a syringe to gently draw out the plasma layer, and then use sterile tools (such as tweezers or a scraper) to carefully remove the middle PRF gel layer to avoid destroying the fibrin network structure, and then freeze-dry and grind to make PRF freeze-dried powder.

[0038] (3) Synthesis of quercetin-modified zeolitic imidazolate framework nanoparticles ZIF-8@QCT

[0039] At room temperature, 300 mg of zinc nitrate (Zn(NO3)2·6H2O) was dissolved in 10 mL of deionized water, and 660 mg of 2-methylimidazole and 8 mg of quercetin were dissolved in 20 mL of methanol solution. The zinc solution was then mixed with the quercetin solution and reacted for 30 minutes under magnetic stirring until the powder was completely dissolved. Finally, the yellow product obtained after centrifugation at 10,000 rpm for 15 minutes was washed several times with methanol and dried overnight in a vacuum atmosphere at 37°C to obtain ZIF-8@QCT.

[0040] (4) Preparation of oxidized dextran ODex:

[0041] Weigh 5 g of dextran and dissolve it in 250 mL of water. After dissolving in a 60°C water bath, cool to room temperature, add 5 g of sodium periodate, react for 3.5 h in the dark, add 3 mL of ethylene glycol to terminate the reaction, continue stirring for 1 h, dialyze with an 8000 dialysis bag, and collect by lyophilization.

[0042] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0043] Example 1

[0044] 0.05 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.01 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0045] 0.05 g of ODex was added to 0.5 mL of deionized water, and 0.005 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0046] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0047] Example 2

[0048] 0.05 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath environment to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0049] 0.05 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0050] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0051] Example 3

[0052] 0.1 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0053] 0.05 g of ODex was added to 0.5 mL of deionized water, and 0.005 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0054] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0055] Example 4

[0056] 0.15 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.01 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0057] 0.05 g of ODex was added to 0.5 mL of deionized water, and 0.005 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0058] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0059] Example 5

[0060] 0.1 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0061] 0.1 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0062] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0063] Example 6

[0064] 0.1 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0065] 0.15 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0066] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0067] Example 7

[0068] 0.15 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0069] 0.1 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0070] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0071] Example 8

[0072] 0.15 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.03 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0073] 0.15 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0074] The Chs-ADH@PRF hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex@ZIF-8@QCT.

[0075] Comparative Example 1

[0076] Dissolve 0.1 g of Chs-ADH in 0.5 mL of deionized water to obtain a first gel solution—Chs-ADH hydrogel solution;

[0077] 0.15 g of ODex was added to 0.5 mL of deionized water, and 0.01 g of ZIF-8@QCT nanoparticles were added to the above solution and stirred for 10 minutes to obtain the second gel solution - ODex@ZIF-8@QCT hydrogel solution;

[0078] The Chs-ADH hydrogel solution was mixed with the ODex@ZIF-8@QCT hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH / ODex@ZIF-8@QCT.

[0079] Comparative Example 2

[0080] 0.15 g of Chs-ADH was dissolved in 0.5 mL of deionized water, and 0.01 g of PRF lyophilized powder was added to the Chs-ADH hydrogel solution in an ice bath to obtain the first gel solution - Chs-ADH@PRF hydrogel solution;

[0081] 0.15 g of ODex was added to 0.5 mL of deionized water and stirred for 10 minutes to obtain the second gel solution—ODex hydrogel solution;

[0082] The Chs-ADH@PRF hydrogel solution was mixed with the ODex hydrogel solution, and the pH was adjusted to 7-8 to obtain the injectable hydrogel Chs-ADH@PRF / ODex.

[0083] Comparative Example 3

[0084] Absorbent gelatin sponge produced by Jiangxi Xiangen Medical Technology Development Co., Ltd. (specification: type B 20mm×20mm×50mm).

[0085] The components of the hydrogel samples of the above examples and comparative examples are shown in Table 1.

[0086] Table 1 Composition of hydrogels in different embodiments and comparative examples

[0087]

[0088] Performance Test:

[0089] 1. Viscosity test

[0090] Test method: The hydrogel was injected onto the test sample stage of the rotational rheometer, the test temperature was set to 37°C, the shear frequency was set to 1 Hz, and the viscosity of the liquid was tested using a stainless steel lamina. The results are shown in Table 2.

[0091] Table 2 Viscosity test results

[0092]

[0093] Result explanation: It can be seen from Table 2 that with the increase of Chs-ADH and ODex content, the viscosity of the hydrogel gradually increases. A hydrogel with too low viscosity may flow too fast in the body, affecting its durability and drug release control in the body. Therefore, it is better when the content of Chs-ADH and ODex is greater than or equal to 10%, but too much content will cause the viscosity to be too high and the hydrogel will block the injection needle, resulting in difficulty in injection. Therefore, preferably, the content of Chs-ADH and ODex is more suitable at 10-15%, and optimally, Chs-ADH: ODex = 1-1.5: 1 (weight ratio), and the viscosity obtained is more suitable. Optimally, the hydrogel synthesized in the ratio of 15% Chs-ADH to 10% ODex in Example 7 is the most stable and smooth to inject, with the attached Figure 1 Schematic diagram of hydrogel injection in Example 7.

[0094] 2. Degradation performance test

[0095] Test method: First, freeze-dry the Chs-ADH@PRF / ODex@ZIF-8@QCT microgel assembly prepared in Example 7 at -80°C. Weigh and record the initial weight of the hydrogel after freeze-drying (the gelatin sponge is consistent with its mass), recorded as W0. Place the prepared freeze-dried hydrogel sample and gelatin sponge in a 24-well plate, add a fixed volume of collagenase solution with a pipette to completely immerse the hydrogel sample, and place it in a 37°C constant temperature shaker. Take out the samples after an interval of 12 hours, discard the excess degradation solution, freeze-dry, weigh, and record the weight, recorded as W t The degradation rate is calculated according to the following formula:

[0096]

[0097] Grouping Degradation rate (%) Example 1 61.20±2.73 Example 2 56.25±2.09 Example 3 59.42±2.52 Example 4 58.85±2.35 Example 5 53.89±1.49 Example 6 51.29±2.77 Example 7 49.16±2.07 Example 8 48.72±2.84 Comparative Example 1 52.23±1.51 Comparative Example 2 59.12±2.87 Comparative Example 3 62.51±1.78

[0098] Results show that in the hydrogel system of the present invention, the amount of Chs-ADH and ODex affects the degradation performance by affecting the cross-linked network structure of the hydrogel network. The more Chs-ADH and ODex content, the better the anti-degradation property of the gel; at the same time, the content of ZIF-8@QCT and PRF also has a great influence on the anti-degradation property of the hydrogel. On the whole, in Examples 5-8 of the present invention, when there are 10-15% Chs-ADH, 10-15% ODex, 1% ZIF-8@QCT, and 3% PRF, the hydrogel has good stability and anti-degradation performance relative to commercially available gelatin sponges, and can better adapt to the complex oral environment.

[0099] 3. Antioxidant capacity test

[0100] Test method: The antioxidant performance of different embodiments and comparative examples was evaluated by scavenging 1,1-diphenyl-2-pyridylhydrazide (DPPH) free radicals. The mixture of different embodiments and comparative examples and DPPH reagent was stirred and incubated in the dark for 30 minutes, and the remaining DPPH was analyzed by UV-visible spectroscopy. The scavenging rate of DPPH was determined as follows:

[0101] D(%)=[[A 空白 -(A 测定 -A 对照 )] / A 空白 ]×100%

[0102] Table 4 DPPH free radical scavenging rate (%)

[0103]

[0104] Results: As shown in Table 4, the efficiency of scavenging free radicals increases with the increase of ZIF-8@QCT, indicating that ZIF-8@QCT can effectively scavenge free radicals and has a good antioxidant effect. In addition, it can be seen that PRF can promote ZIF-8@QCT to exert its antioxidant effect to a certain extent, and has a certain synergistic effect with ZIF-8@QCT.

[0105] 4. In vitro coagulation test

[0106] Test method: The in vitro whole blood coagulation index was used to study the coagulation-promoting effect of the hemostatic material. 100 μL of hydrogel was placed in a clean EP tube. 100 μL of anticoagulated blood and 20 μL of CaCl2 (0.2 mol / L) solution were added in sequence and mixed evenly. Then, the coagulation degree of the blood in the centrifuge tube was observed at 37°C.

[0107] Table 5 In vitro whole blood coagulation index

[0108]

[0109]

[0110] Results: As can be seen from Table 5, the addition of PRF has a very obvious coagulation effect. When PRF comes into contact with blood, the growth factors in platelets are activated and released, which helps promote local tissue repair and regeneration. The release of platelet factors enhances the responsiveness of PRF to blood and damaged tissues to a certain extent; PRF contains a high concentration of fibrin, which can quickly form a mesh structure, which helps local blood coagulation and hemostasis. In addition, the content of Chs-ADH and ODex also has a great influence on the coagulation index, which may be because the higher content of Chs-ADH and ODex can form a more dense and stable three-dimensional network structure, which helps to capture blood cells, platelets and coagulation factors, promote their aggregation and stability, and thus accelerate the occurrence of coagulation.

[0111] 5. In vitro antibacterial test

[0112] Test method: Porphyromonas gingivalis, Escherichia coli and Staphylococcus aureus were revived and cultured to the logarithmic growth phase, centrifuged, and the bacteria were collected and resuspended in physiological saline. The final bacterial solution concentration was 1×10 8 CFU / mL. After adding 500μL of the corresponding hydrogel solution to a 24-well plate, UV light was used to solidify the solution. 500μL of saline was added to the control group. Then, 100μL of the diluted bacterial suspension and 1mL of sterile saline were added to each well, incubated at 37°C for 24 hours, and then spread on BA medium or LB agar plates after gradient dilution and counted.

[0113] Table 6 In vitro antibacterial rate (%)

[0114]

[0115]

[0116] The results show that ZIF-8@QCT is the main antibacterial component, and the higher the addition amount, the stronger the antibacterial effect. Chs-ADH@PRF and ODex are polysaccharides and bioactive factors, which have no obvious effect on killing bacteria.

[0117] In summary, it can be seen that the hydrogel of the embodiment of the present invention uses Chs-ADH and ODex as matrix materials, introduces PRF and ZIF-8@QCT, endows the hydrogel with antibacterial, antioxidant and immunomodulatory functions, and optimizes the distribution ratio of each group to obtain a hydrogel with good injectability, stability and anti-degradation properties, so that it can still maintain effective coagulation function in the oral environment with high humidity and high enzyme activity. It is particularly suitable for oral wounds.

[0118] The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, all possible combinations of the technical features in the above-mentioned embodiments are not described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims.

Claims

1. An injectable hydrogel for rapid oral hemostasis and antibacterial properties, characterized in that: The invention comprises the following components in weight percentage: 5-15% of amino chondroitin sulfate, 5-15% of oxidized dextran, 1-3% of platelet-rich fibrin, and 0.5-1% of quercetin-modified zeolite imidazole acid framework nanoparticles.

2. The injectable hydrogel according to claim 1, characterized in that The invention comprises the following raw materials in weight percentage: 10-15% of amino chondroitin sulfate, 10-15% of oxidized dextran, 1-3% of platelet-rich fibrin, and 0.5-1% of quercetin-modified zeolite imidazole acid framework nanoparticles.

3. The injectable hydrogel according to any one of claims 1 to 2, characterized in that According to the weight ratio, the amino chondroitin sulfate: oxidized dextran = 1-1.5:

1.

4. The injectable hydrogel according to claim 1, characterized in that The quercetin-modified zeolite imidazolate framework nanoparticles are prepared by the following method, including: dissolving a soluble zinc salt in deionized water at room temperature to obtain a zinc solution; dissolving 2-methylimidazole and quercetin in methanol to obtain a quercetin solution; then mixing the zinc solution with the quercetin solution, stirring and reacting until the powder is completely dissolved; finally, centrifuging the obtained yellow product, washing it with methanol for several times, and vacuum drying it at 30-40° C. to obtain the quercetin-modified zeolite imidazolate framework nanoparticles.

5. The injectable hydrogel according to claim 4, characterized in that The zinc solution contains Zn 2+ The concentration is 0.08-0.15 mol / L. In the quercetin solution, the concentration of 2-methylimidazole is 0.40-0.50 mol / L, the concentration of quercetin is 1.325 mmol / L, and the volume ratio of the zinc solution to the quercetin solution is 1:

2.

6. The injectable hydrogel according to claim 1, characterized in that The aminated chondroitin sulfate is prepared by the following method, including: dissolving sodium chondroitin sulfate and diacyl adipic acid in deionized water to obtain solution I; dissolving 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 1-hydroxybenzotriazole hydrate in a dimethyl sulfoxide / water mixture to obtain solution II; then dropping solution II into solution I, adjusting the pH to 5.0, and reacting at room temperature; after the reaction is completed, dropping ethanol to precipitate, dissolving the precipitate in water, dialyzing the solution in pure water, the dialysis molecular weight cutoff is 500Da, and then freeze-drying to obtain the aminated chondroitin sulfate.

7. The injectable hydrogel according to claim 6, characterized in that The dimethyl sulfoxide / water mixture is a mixture of dimethyl sulfoxide and water in a volume ratio of 1:1; sodium chondroitin sulfate: diacyl adipic acid: deionized water: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloric acid: 1-hydroxybenzotriazole hydrate: dimethyl sulfoxide / water mixture = 0.5g: 10g: 100mL: 0.8g: 0.7g: 10mL.

8. The method for preparing an injectable hydrogel according to any one of claims 1 to 7, characterized in that: include: Dissolving the aminated chondroitin sulfate in a first solvent to obtain an aminated chondroitin sulfate hydrogel solution, and then adding platelet-rich fibrin lyophilized powder to the aminated chondroitin sulfate hydrogel solution in an ice bath environment to obtain a first hydrogel solution; adding oxidized dextran to a second solvent to obtain an oxidized dextran solution, and then adding quercetin-modified zeolite imidazolate framework nanoparticles to the oxidized dextran solution for dispersion to obtain a second hydrogel solution; The first hydrogel solution and the second hydrogel solution are mixed, and the pH is adjusted to 7-8 to obtain the injectable hydrogel.

9. The preparation method according to claim 8, characterized in that: The first solvent and the second solvent are any one of ultrapure water, PBS buffer, and physiological saline.

10. A wound dressing, characterized in that: The invention comprises the injectable hydrogel according to any one of claims 1 to 7, or the injectable hydrogel prepared by the preparation method according to any one of claims 8 to 9.

Images