Self-repairing peach gum hydrogel based on reversible subject-object and boric acid ester bond, preparation method and application

Abstract

The invention belongs to the technical field of biomedical materials and biomedical engineering, and discloses self-repairing peach gum hydrogel based on reversible subject-object and boric acid ester bonds, a preparation method and application. The preparation method comprises the following steps: modifying boric acid and amantadine on purified peach gum to obtain boric acid-amantadine peach gum; modifying chitosan with carboxymethyl cyclodextrin to obtain beta cyclodextrin-chitosan; boric acid-amantadine peach gum and beta cyclodextrin-chitosan are mixed, and the self-repairing peach gum hydrogel is obtained through the mutual synergistic effect of a subject, a guest and boric acid ester. The hydrogel has multiple functions of excellent biocompatibility, self-repairing, hemostasis, antibiosis and the like, and can be applied to body surface skin wound repairing.

Description

Technical Field

[0001] This invention belongs to the fields of biomedical materials and biomedical engineering technology, and relates to a self-healing gum hydrogel based on host-guest dual borate ester bonds, its preparation method and application. Background Technology

[0002] Traumatic bleeding, especially massive hemorrhage caused by damage to solid organs such as the liver and heart, is one of the leading causes of death in clinical emergency care. Such trauma is characterized by rapid bleeding, difficulty in hemostasis, and a high risk of hemorrhagic shock, placing extremely high demands on the rapid hemostatic performance, biocompatibility, and tissue compatibility of hemostatic materials. Currently, commonly used hemostatic materials include gelatin sponges, chitosan dressings, and hemostatic powders, but these have certain limitations: some materials have insufficient hemostatic rates and are inadequate for arterial bleeding; some synthetic materials have poor biocompatibility, easily triggering inflammatory reactions or tissue rejection; while natural materials have better compatibility, their weak mechanical strength and insufficient adhesion make them prone to detachment from moist tissue surfaces, affecting hemostatic efficacy. Hydrogel-based hemostatic materials, due to their three-dimensional network structure, can rapidly absorb water from the blood, concentrate clotting factors, and promote coagulation through physical obstruction and bioactivity, while also adhering closely to the wound surface, making them a research hotspot in the field of trauma hemostasis. However, existing hydrogels still need improvement in terms of rapid hemostasis efficiency, biocompatibility, and adaptability to complex organ injuries. Developing hydrogel materials that combine high hemostasis efficiency, good biocompatibility, and tissue adhesion is of great significance for improving the success rate of trauma treatment. Summary of the Invention

[0003] The purpose of this invention is to provide a method for preparing and applying a self-healing gum hydrogel based on host-guest boronic acid ester bonds. The hydrogel prepared by this method has excellent hemostatic, self-healing, and antibacterial properties, and also has good biocompatibility.

[0004] In a first aspect, the present invention provides a method for preparing a self-healing gum hydrogel based on reversible host-guest and borate ester bonds, comprising:

[0005] S1. The purified peach gum was added to PBS buffer, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide were added for activation. Then, 3-aminophenylborate salt and adamantane were added for reaction. After the reaction was completed, the mixture was dialyzed and freeze-dried to obtain adamantane-boric acid double-modified peach gum.

[0006] S2. Dissolve sodium carboxymethyl-β-cyclodextrin in distilled water, then add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide for activation, then add chitosan acetate solution for reaction, and freeze-dry after reaction to obtain chitosan / carboxymethyl-β-cyclodextrin.

[0007] S3. Dissolve the adamantane-boric acid dual-modified peach gum in PBS buffer to obtain an adamantane-boric acid dual-modified peach gum solution; dissolve chitosan / carboxymethyl-β-cyclodextrin in acetic acid solution to obtain a chitosan / carboxymethyl-β-cyclodextrin solution; mix the adamantane-boric acid dual-modified peach gum solution with the chitosan / carboxymethyl-β-cyclodextrin solution and adjust the pH to 8.0 to obtain a self-healing peach gum hydrogel.

[0008] In some embodiments of the present invention, the volume concentration of the acetic acid solution is 1% (v / v).

[0009] In some embodiments of the present invention, in step S1, the mass ratio of purified gum arabic, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 1000:(950-970):(470-950).

[0010] In some embodiments of the present invention, in step S1, the molar ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC•HCl) to N-hydroxysuccinimide (NHS) is (1-2):1.

[0011] In some embodiments of the present invention, in step S1, the mass ratio of purified gum arabic, 3-aminophenylborate salt to adamantane is 1000:(260-290):(258-288).

[0012] In some embodiments of the present invention, in step S1, the activation time is 15 to 30 minutes, and the reaction time is 8 to 12 hours.

[0013] In some embodiments of the present invention, in step S2, the molar ratio of sodium carboxymethyl-β-cyclodextrin, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 250:(300-330):(160-300).

[0014] In some embodiments of the present invention, in step S2, the mass ratio of sodium carboxymethyl-β-cyclodextrin salt to chitosan in the chitosan acetate solution is 1:1.

[0015] In some embodiments of the present invention, in step S3, the mass ratio of adamantane-boric acid double-modified peach gum to chitosan / carboxymethyl-β-cyclodextrin is 25:(20-30).

[0016] In some embodiments of the present invention, in step S3, the ratio of the adamantane-boric acid double-modified peach gum to PBS buffer is 25 mg: (200-300) μL; the ratio of the chitosan / carboxymethyl-β-cyclodextrin to acetic acid solution is 25 mg: (200-300) μL.

[0017] In a second aspect, the present invention provides a self-healing gum hydrogel based on reversible host-guest and borate ester bonds prepared by the above-described preparation method.

[0018] A third aspect of the present invention provides the application of the above-mentioned self-healing gum hydrogel based on reversible host-guest and borate ester bonds in the preparation of biological tissue hemostatic materials or biological tissue adhesion materials.

[0019] Compared with existing technologies, this invention is the first to combine reversible host-guest interactions and borate ester bonds, and synthesizes a self-healing gum hydrogel based on host-guest and borate ester bonds in one step through a simple and mild amide reaction, thus realizing its multifunctional application. Attached Figure Description

[0020] To more clearly illustrate the technical solutions of the present invention, the accompanying drawings used in the description of the embodiments or prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0021] Figure 1 This is a schematic diagram of the self-healing peach gum hydrogel prepared in Example 1; wherein, Figure A is a schematic diagram of the hydrogel being cut open; Figure B is a schematic diagram of the healing of the moisturizing gel after 40 seconds; and Figure C is a schematic diagram of the self-healing gel being lifted with tweezers to verify its self-healing effect.

[0022] Figure 2 This is a schematic diagram of the hemostatic effect of the self-healing gum hydrogel prepared in Example 1 on the liver. In Figure A, the liver wound without the use of the hydrogel is shown; and in Figure B, the liver wound with the use of the hydrogel is shown.

[0023] Figure 3 Figure A shows the antibacterial effect of the self-healing gum hydrogel prepared in Example 2. Figure B shows the coating culture results after co-incubation of the hydrogel with Escherichia coli; Figure C shows the coating culture results after co-incubation of the hydrogel with Escherichia coli without using the hydrogel; Figure D shows the coating culture results after co-incubation of the hydrogel with Staphylococcus aureus without using the hydrogel. Detailed Implementation

[0024] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0025] Preparation of purified peach gum in this embodiment of the invention:

[0026] Dissolve 10g of natural peach gum in 500mL of distilled water, add 16g of sodium hydroxide, stir in a water bath at 60℃ for 2h, and filter under reduced pressure; add 33mL of concentrated hydrochloric acid (0.012 mol / mL) to the filtrate, dialyze with distilled water for 3 days, and freeze dry to obtain purified peach gum.

[0027] Example 1

[0028] 1) Weigh 1000 mg of purified peach gum and dissolve it in 100 mL of PBS buffer (pH 7.4). Add 950 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC•HCl) and 470 mg of N-hydroxysuccinimide (NHS) to activate the peach gum solution for 15 min. Then add 260 mg of 3-aminophenylborate and 258 mg of adamantane and react for 12 h. Dialyze for 3 days and freeze-dry to obtain borate-adamantane double-modified peach gum.

[0029] 2) Dissolve 250 mg of chitosan in 20 mL of 1% (v / v) acetic acid solution; dissolve 250 mg of sodium carboxymethyl-β-cyclodextrin in 21 mL of distilled water; add 300 mg of EDC•HCl and 160 mg of NHS to the sodium carboxymethyl-β-cyclodextrin aqueous solution, activate for 15 min, then add chitosan acetic acid solution, mix well, react for 8 h, freeze dry to obtain chitosan / carboxymethyl-β-cyclodextrin.

[0030] 3) Take 25 mg of adamantane-boric acid double-modified peach gum and dissolve it in 200 µL of PBS buffer (pH 7.4); take 20 mg of chitosan / carboxymethyl-β-cyclodextrin and dissolve it in 200 μL of acetic acid; mix the two solutions and adjust the pH to 8.0 to obtain a self-healing peach gum hydrogel based on host-guest dual borate ester bond.

[0031] Example 2

[0032] 1) Weigh 1000 mg of purified peach gum and dissolve it in 100 mL of PBS buffer (pH 7.4). Add 962 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC•HCl) and 577 mg of N-hydroxysuccinimide (NHS) to activate the peach gum solution for 15 min. Then add 271 mg of 3-aminophenylborate and 260 mg of adamantane and react for 12 h. Dialyze for 3 days and freeze-dry to obtain borate-adamantane double-modified peach gum.

[0033] 2) Dissolve 250 mg of chitosan in 25 mL of 1% (v / v) acetic acid solution; dissolve 250 mg of sodium carboxymethyl-β-cyclodextrin in 25 mL of distilled water; add 320 mg of EDC•HCl and 198 mg of NHS to the sodium carboxymethyl-β-cyclodextrin aqueous solution, activate for 15 min, then add chitosan acetic acid solution, mix well, react for 8 h, freeze dry to obtain chitosan / carboxymethyl-β-cyclodextrin.

[0034] 3) Take 25 mg of adamantane-boric acid double-modified peach gum and dissolve it in 250 µL of PBS buffer (pH 7.4); take 25 mg of chitosan / carboxymethyl-β-cyclodextrin and dissolve it in 250 μL of acetic acid; mix the two solutions and adjust the pH to 8.0 to obtain a self-healing peach gum hydrogel based on host-guest dual borate ester bond.

[0035] Example 3

[0036] 1) Weigh 1000 mg of purified peach gum and dissolve it in 100 mL of PBS buffer (pH 7.4). Add 970 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC•HCl) and 950 mg of N-hydroxysuccinimide (NHS) to activate the peach gum solution for 15 min. Then add 290 mg of 3-aminophenylborate and 288 mg of adamantane and react for 12 h. Dialyze for 3 days and freeze-dry to obtain borate-adamantane double-modified peach gum.

[0037] 2) Dissolve 250 mg of chitosan in 25 mL of 1% (v / v) acetic acid solution; dissolve 250 mg of sodium carboxymethyl-β-cyclodextrin in 25 mL of distilled water; add 330 mg of EDC•HCl and 300 mg of NHS to the sodium carboxymethyl-β-cyclodextrin aqueous solution, activate for 15 min, then add chitosan acetic acid solution, mix well, react for 8 h, freeze dry to obtain chitosan / carboxymethyl-β-cyclodextrin.

[0038] 3) Take 25 mg of adamantane-boric acid double-modified peach gum and dissolve it in 300 µL of PBS buffer (pH 7.4); take 30 mg of chitosan / carboxymethyl-β-cyclodextrin and dissolve it in 300 μL of acetic acid; mix the two solutions and adjust the pH to 8.0 to obtain a self-healing peach gum hydrogel based on host-guest dual borate ester bond.

[0039] Test Example 1

[0040] The self-healing ability of gum arabic hydrogel based on host-guest dual borate ester bonds was verified, and the results are as follows: Figure 1 As shown.

[0041] The self-healing gum hydrogel based on host-guest and borate ester bonds obtained in Example 1 was cut into two equal parts using a scalpel, as follows: Figure 1 As shown in Figure A; then the separate self-healing gum hydrogels based on host-guest and borate ester bonds are assembled together, and after about 40 seconds, as shown in Figure A; Figure 1 As shown in Figure B, the separated gum arabic undergoes self-healing; then, the self-healed gum arabic hydrogel based on host-guest and borate ester bonds is lifted with tweezers, as shown in Figure B. Figure 1 As shown in Figure C, the healed gum hydrogel based on host-guest boronic acid ester bonds remained intact after being lifted, indicating that self-repair occurred.

[0042] Test Example 2

[0043] The hemostatic ability of gum arabic hydrogel based on host-guest dual borate ester bonds was verified, and the results are as follows: Figure 2 As shown.

[0044] Eight-week-old female SD rats weighing 209g were anesthetized with 20% urethane. The abdominal cavity was opened with dissecting scissors, and the left lobe of the liver was pulled out with forceps. A filter paper was placed underneath to monitor the amount of bleeding. A 1 cm long and 2 mm deep incision was made on the left lobe with a scalpel. (The control group was as follows...) Figure 2 As shown in Figure A, bleeding continued for one minute, with bloodstains spreading continuously on the filter paper; after the wound was plugged with the self-healing gum arabic hydrogel based on host-guest dual borate ester bonds obtained in Example 1, the hemostatic effect was as follows: Figure 2 As shown in Figure B, the liver bleeding stopped within 70 seconds, and there was no blood spreading on the filter paper, indicating that the liver wound bleeding was low and the hemostasis was successful when using the self-healing gum hydrogel based on host-guest boronic acid ester bonds.

[0045] Test Example 3

[0046] The antibacterial activity of the self-healing gum hydrogel based on host-guest and borate ester bonds was tested, and the results are as follows: Figure 3 As shown.

[0047] 100 μL of *E. coli* and *Staphylococcus aureus* bacterial suspension cultured to an optical density of 0.7 was diluted 100,000 times and plated on agar plates. The mixture was incubated at 37°C for 24 hours. The self-healing gum arabic hydrogel based on host-guest and borate ester bonds obtained in Example 2 was co-cultured with 100 μL of *E. coli* and *Staphylococcus aureus* bacterial suspension cultured to an optical density of 0.7 for 1 hour. The co-cultured bacterial suspension was diluted 100,000 times and plated on agar plates. The mixture was then incubated at 37°C for 24 hours. Figure 3 As shown in Figures A and C, no colonies of *E. coli* and *Staphylococcus aureus* co-cultured with the host-guest boronic acid ester self-healing gum hydrogel grew on the plates, while a large number of colonies of *E. coli* and *Staphylococcus aureus* without co-culture grew on the plates. This indicates that the host-guest boronic acid ester self-healing gum hydrogel can kill the co-cultured *E. coli* and *Staphylococcus aureus*, and has antibacterial properties.

[0048] While some embodiments of the present general inventive concept have been shown and described, those skilled in the art will understand that changes may be made to these embodiments without departing from the principles and spirit of the present general inventive concept, the scope of which is defined by the claims and their equivalents.

Claims

1. A method for preparing self-healing gum hydrogel based on reversible host-guest and borate ester bonds, characterized in that, The preparation method includes the following steps: S1. The purified peach gum was added to PBS buffer, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide were added for activation. Then, 3-aminophenylborate salt and adamantane were added for reaction. After the reaction was completed, the mixture was dialyzed and freeze-dried to obtain adamantane-boric acid double-modified peach gum. S2. Dissolve sodium carboxymethyl-β-cyclodextrin in distilled water, then add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide for activation, then add chitosan acetate solution for reaction, and freeze-dry after reaction to obtain chitosan / carboxymethyl-β-cyclodextrin. S3. Dissolve the adamantane-boric acid dual-modified peach gum in PBS buffer to obtain an adamantane-boric acid dual-modified peach gum solution; dissolve chitosan / carboxymethyl-β-cyclodextrin in acetic acid solution to obtain a chitosan / carboxymethyl-β-cyclodextrin solution; mix the adamantane-boric acid dual-modified peach gum solution with the chitosan / carboxymethyl-β-cyclodextrin solution and adjust the pH to 8.0 to obtain a self-healing peach gum hydrogel.

2. The preparation method according to claim 1, characterized in that, In step S1, the mass ratio of purified peach gum, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 1000:(950-970):(470-950).

3. The preparation method according to claim 1, characterized in that, In step S1, the mass ratio of purified gum arabic, 3-aminophenylborate salt to adamantane is 1000:(260-290):(258-288).

4. The preparation method according to claim 1, characterized in that, In step S1, the activation time is 15 to 30 minutes, and the reaction time is 8 to 12 hours.

5. The preparation method according to claim 1, characterized in that, In step S2, the molar ratio of sodium carboxymethyl-β-cyclodextrin, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and N-hydroxysuccinimide is 250:(300-330):(160-300).

6. The preparation method according to claim 1, characterized in that, In step S2, the mass ratio of sodium carboxymethyl-β-cyclodextrin salt to chitosan in the chitosan acetate solution is 1:

1.

7. The preparation method according to claim 1, characterized in that, In step S3, the mass ratio of adamantane-boric acid double-modified peach gum to chitosan / carboxymethyl-β-cyclodextrin is 25:(20-30).

8. The preparation method according to claim 7, characterized in that, In step S3, the ratio of the adamantane-boric acid double-modified peach gum to PBS buffer is 25 mg: (200-300) μL; the ratio of the chitosan / carboxymethyl-β-cyclodextrin to acetic acid solution is 25 mg: (200-300) μL.

9. The self-healing gum hydrogel based on reversible host-guest and borate ester bonds prepared by any one of the preparation methods according to claims 1-8.

10. The application of the self-healing gum hydrogel based on reversible host-guest and borate ester bonds as described in claim 9 in the preparation of biological tissue hemostatic materials or biological tissue adhesion materials.

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