Antibacterial tissue patch with excellent adhesion as well as preparation method and application of antibacterial tissue patch

Abstract

The invention relates to an antibacterial tissue patch with excellent adhesion as well as a preparation method and application of the antibacterial tissue patch. The antibacterial tissue patch is formed by alternately coating at least one carrier layer and at least one antibacterial tissue adhesion layer, the carrier layer is a film-forming polymer, the antibacterial tissue adhesion layer is an antibacterial copolymer, and the antibacterial copolymer is poly (N-vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopamine quinone acrylate). The preparation method comprises the following steps: dissolving the film-forming polymer in an organic solvent, coating, and drying to obtain the carrier layer; then preparing an antibacterial tissue adhesion layer, and then coating the tissue adhesion layer with the carrier layer to form a multi-layer alternating structure which is excellent in antibacterial property, high in biocompatibility and suitable for functional composite materials.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to an antibacterial tissue patch with excellent adhesion, a preparation method and application thereof. Background Art

[0002] Bacterial infection has a significant impact in the medical field and poses a serious threat to human health. Medical implant materials will have a 4% to 10% implant failure rate due to bacterial infection in the early stage of implantation. Nowadays, the most common antibacterial method is to use antibacterial agents. However, the use of these compounds has led to the formation of drug-resistant microorganisms. Therefore, the development of medical materials with intrinsic antibacterial properties has become one of the general trends in the medical field.

[0003] Dopamine is a catecholamine neurotransmitter in the human body. Reactive oxygen species (ROS) are produced during the oxidation of catechol groups, and ROS have been shown to be an effective broad-spectrum bactericide in many industrial and biomedical applications. After being oxidized to quinones, catechol groups can react with amine groups to form Schiff base reactions. In the medical field, Schiff bases have antibacterial, bactericidal, anti-tumor and antiviral biological activities.

[0004] In addition, the current means of achieving wound closure or implant fixation in surgical operations mainly use traditional invasive techniques such as sutures, staples, and rivets, which can easily cause secondary injuries and suture leakage, resulting in a long wound healing time, poor healing quality, and many complications. Therefore, tissue adhesives and hydrogels are often used as substitutes to achieve wound adhesion.

[0005] However, tissue adhesives and hydrogels also have the following problems: (1) They may overswell, leading to serious medical complications, such as local nerve compression, and the increase in water content will also lead to a decrease in cross-linking density, affecting the mechanical properties and stability of adhesives and hydrogels; (2) The molecular chain size is large and the tissue surface penetration ability is weak, resulting in unsatisfactory adhesion, insufficient interface adhesion, tissue leakage, air leakage, and other situations, which in turn cause inflammation, wound infection, etc.; (3) The cohesive strength between hydrogels and tissue adhesives and tissues does not match, resulting in premature failure of the hydrogel itself. The above shortcomings limit the use of hydrogels and tissue adhesives. Summary of the invention

[0006] The purpose of the present invention is to provide an antibacterial tissue patch with excellent adhesion and a preparation method and application thereof, which solves the problem of low adhesion and antibacterial properties of tissue patches in the prior art.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is: The first aspect of the present invention provides an antibacterial tissue patch with excellent adhesion, which is formed by alternately coating at least one carrier layer and at least one antibacterial tissue adhesion layer; the carrier layer is a film-forming polymer, the antibacterial tissue adhesion layer is an antibacterial copolymer, and the antibacterial copolymer is poly (N-vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopaminequinone acrylate).

[0008] Furthermore, the film-forming polymer is a mixture of one or more selected from polylactic acid, polyglycolic acid, polycaprolactone and polyhydroxyalkanoate.

[0009] Furthermore, the film-forming polymer is a lactic acid-glycolic acid copolymer.

[0010] Furthermore, the antibacterial copolymer is formed by copolymerizing dopamine, acrylic acid and N-vinyl pyrrolidone and then inducing partial oxidation. The molar ratio of dopamine, acrylic acid and N-vinyl pyrrolidone is 0.6-1.4:1:1.

[0011] A second aspect of the present invention provides a method for preparing an antibacterial tissue patch with excellent adhesion, comprising the following steps: (1) dissolving the film-forming polymer, coating it, and drying it to obtain a carrier layer; (2) Under a nitrogen atmosphere, acrylic acid, dopamine hydrochloride and a condensing agent are subjected to condensation esterification to prepare dopamine acrylate; (3) adding N-vinyl pyrrolidone and a photoinitiator to the dopamine acrylate, stirring evenly under light-proof conditions, and then polymerizing under ultraviolet light, and then standing for purification to obtain poly(N-vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopaminequinone acrylate); (4) The poly(N-vinylpyrrolidone-acrylic acid-dopamine acrylate-dopaminequinone acrylate) is dissolved, and then coated on the carrier layer and dried, and the coating and drying processes are repeated to obtain the antibacterial tissue patch with excellent adhesion.

[0012] Furthermore, in step (2), the molar ratio of the acrylic acid to the condensation agent is 1:2.

[0013] Furthermore, the condensing agent in step (2) is a mixed solution of EDC and NHS, and the molar ratio of the EDC to the NHS is 1:1.

[0014] Furthermore, the reaction temperature in step (2) is room temperature, and the reaction time is 6 to 18 hours.

[0015] Furthermore, in step (3), the photoinitiator is a mixture of one or more selected from TPO, MBF and BP, and the molar ratio of the dopamine acrylate, the N-vinyl pyrrolidone and the photoinitiator is 1:1:0.04.

[0016] Furthermore, in step (3), the UV polymerization time is 10 to 30 minutes, and the standing time is 24 to 72 hours.

[0017] The third aspect of the present invention provides an application of the above-mentioned antibacterial tissue patch with excellent adhesion in a functional composite material.

[0018] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art: The present invention integrates the carrier layer and the antibacterial tissue adhesion layer, and forms a non-covalent interaction with the tissue by using a polymer material with a catechol group and a quinone group, and the quinone group forms a covalent bond with the amine group on the tissue surface, which can not only adhere to the tissue surface immediately but also provide long-term adhesion. At the same time, it has good biological activities such as antibacterial, bactericidal and anti-tumor. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Hereinafter, some specific embodiments of the present invention will be described in detail in an exemplary and non-limiting manner with reference to the accompanying drawings. The same reference numerals in the accompanying drawings indicate the same or similar components or parts. It should be understood by those skilled in the art that these drawings are not necessarily drawn to scale. In the accompanying drawings: Figure 1 This is an infrared absorption spectrum of the antibacterial tissue patch with excellent adhesion prepared in Example 1 provided in the present invention. DETAILED DESCRIPTION

[0020] The present invention provides an antibacterial tissue patch with excellent adhesion, a preparation method and application thereof. Specifically, the antibacterial tissue patch is formed by alternately coating at least one carrier layer and at least one antibacterial tissue adhesion layer; the carrier layer is a film-forming polymer, and the antibacterial tissue adhesion layer is a copolymer with antibacterial function.

[0021] For the above-mentioned carrier layer, its film-forming polymer is a mixture of one or more selected from polylactic acid, polyglycolic acid, polycaprolactone and polyhydroxyalkanoate. For the above-mentioned antibacterial tissue adhesion layer, it is copolymerized by multiple polymer monomers. In the antibacterial tissue adhesion layer, the component that plays an antibacterial role is a mixture of one or more selected from chitosan, sodium alginate, polylysine, quaternary ammonium salt, imidazole salt, antibiotic, metal / metal oxide, carbon material, dopamine and dopamine quinone; the component that plays an adhesion role is a mixture of one or more selected from succinimide ester, isocyanate, acrylate, maleimide, epoxide, catechol and o-benzoquinone derivatives.

[0022] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] Embodiment 1: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof, comprising the following steps: (1) Preparation of carrier layer 3 g of lactide-glycolide copolymer (the copolymer contains 50% lactide and 50% glycolide, Mw=100 kDa) solid was dissolved in 30 mL of dichloromethane to prepare a 10% w / v solution; Subsequently, a 10% w / v lactide-co-glycolide copolymer solution was coated on a polyethylene plate using a film scraper, and the coating thickness was controlled to be about 10 μm, and the layer was not peeled off from the polyethylene plate. After coating, the layer was placed in a vacuum drying oven and dried at 30° C. for 30 minutes to obtain a carrier layer. (2) Preparation of antibacterial tissue adhesion layer Add the purified acrylic acid monomer (10 g, 0.139 mol) by vacuum distillation to 50 mL DMSO solution, deoxygenate with N2 for 30 min, maintain N2 atmosphere, add dopamine hydrochloride (26.5 g, 0.139 mol) in DMSO (10 mL) under magnetic stirring, stir for 10 min to fully mix the solution, and finally slowly add EDC (26.65, 0.139 mol) in DMSO (70 mL) and NHS (16 g, 0.139 mol) in DMSO (30 mL). Keep the reaction at 25 ° C for 6 h to obtain dopamine acrylate solution; Then, N-vinyl pyrrolidone (15.45 g, 0.139 mol) and a photoinitiator (4% wt acrylic acid and vinyl pyrrolidone, 1.02 g) were added to the aforementioned dopamine acrylate solution, stirred and mixed evenly under light-proof conditions, and polymerized under ultraviolet light for 10 min; The reaction solution was then placed in a fume hood at 50°C for 48 hours to induce oxidation of some catechol groups. After the reaction was completed, the reaction solution was slowly dripped into isopropanol, and a grayish white precipitate was precipitated. Subsequently, it was washed three times with isopropanol / ether (volume ratio of 2:1) and ether, respectively, and dried in a fume hood to obtain a poly(vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopamine quinone acrylate) tetrapolymer, that is, an antibacterial tissue adhesion layer was prepared; (3) Preparation of tissue patches The poly(vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopamine quinone acrylate) quaternary copolymer prepared in step (2) is dissolved in a mixed solvent of dichloromethane / methanol (volume ratio of 2:1) to prepare a 10% w / v solution; The copolymer solution is coated on a polyethylene plate using a film scraper, the coating thickness is controlled to be about 10 µm, and the layer is not peeled off from the polyethylene plate. After coating, the plate is placed in a vacuum drying oven and dried at 30°C for 30 minutes. Then, a layer of the 10% w / v carrier layer solution prepared in step (1) is cast on the antibacterial tissue adhesion layer and subjected to the same drying treatment. Subsequently, the 10% w / v antibacterial tissue adhesion layer prepared in the aforementioned step (2) and the carrier layer solution prepared in the aforementioned step (1) are repeatedly coated and dried, thereby obtaining an antibacterial tissue patch with a multi-layer alternating structure, the thickness of which is controlled to be 40-50 µm.

[0024] Embodiment 2: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The difference between this embodiment and embodiment 1 is only that the molar ratio of acrylic acid monomer to dopamine hydrochloride in step (2) is 1:0.6.

[0025] Embodiment 3: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The only difference between the embodiment 1 and the embodiment 1 is that the molar ratio of acrylic acid monomer to dopamine hydrochloride in step (2) is 1:0.8.

[0026] Embodiment 4: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The difference between this embodiment and embodiment 1 is only that the molar ratio of acrylic acid monomer to dopamine hydrochloride in step (2) is 1:1.2.

[0027] Embodiment 5: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The difference between this embodiment and embodiment 1 is only that the molar ratio of acrylic acid monomer to dopamine hydrochloride in step (2) is 1:1.4.

[0028] Embodiment 6: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The difference between this embodiment and embodiment 1 is only that the reaction solution is left to stand for 24 hours in step (2).

[0029] Embodiment 7: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The only difference between this embodiment and embodiment 1 is that the reaction solution is left to stand for 36 hours in step (2).

[0030] Embodiment 8: This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The only difference between this embodiment and embodiment 1 is that the reaction solution is left to stand for 60 hours in step (2).

[0031] Example 9 This embodiment provides an antibacterial tissue patch with excellent adhesion and a preparation method thereof. The only difference between this embodiment and embodiment 1 is that the reaction solution is left to stand for 72 hours in step (2).

[0032] Comparative Example 1 This embodiment provides an antibacterial tissue patch and a preparation method thereof, which is different from Embodiment 1 only in that dopamine hydrochloride is not used in step (2) to prepare the antibacterial tissue adhesion layer.

[0033] Comparative Example 2 This embodiment provides an antibacterial tissue patch and a preparation method thereof, which is different from Embodiment 1 only in that the reaction solution is left to stand for 120 hours in step (2).

[0034] In the present invention, the antibacterial tissue patches prepared in the above-mentioned embodiments and comparative examples are tested and characterized for their performance. The specific test methods and results of each performance are as follows: (1) Adhesion durability test According to YY / T 0729.3-2009 "Test Method for Adhesive Properties of Tissue Adhesives Part 3: Tensile Strength", the above Examples 1 to 9 and Comparative Example 1 were tested for adhesive strength. The specific description is as follows: Using cyanoacrylate adhesive, a 2.5cm*2.5cm tissue patch and fresh pig skin were bonded to the test substrate respectively. After the cyanoacrylate adhesive was cured, the tissue patch and pig skin were bonded together, and a force of 2N was applied to the bonding area for 5 minutes. The test sample was pulled and separated until it was destroyed using a universal testing machine. The test speed was a constant speed of 10mm / min. The maximum load when the tissue patch and pig skin were destroyed was the bonding strength.

[0035] Table 1 Comparison of bonding strength of various embodiments and comparative examples serial number Composition and reaction time of antibacterial tissue adhesion layer in antibacterial tissue patch Adhesion strength (N) Example 1 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; and the reaction solution in step (2) is allowed to stand for 48 hours. 4.57 Example 2 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:0.6; the reaction solution in step (2) is allowed to stand for 48 hours. 4.04 Example 3 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:0.8; the reaction solution is allowed to stand for 48 hours in step (2). 4.27 Example 4 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1.2; the reaction solution in step (2) is allowed to stand for 48 hours. 4.33 Example 5 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1.4; the reaction solution in step (2) is allowed to stand for 48 hours. 4.28 Example 6 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; the reaction solution in step (2) is allowed to stand for 24 hours. 4.42 Example 7 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; the reaction solution in step (2) is allowed to stand for 36 hours. 4.46 Example 8 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; and the reaction solution in step (2) is allowed to stand for 60 hours. 3.45 Example 9 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; and the reaction solution in step (2) is allowed to stand for 72 hours. 3.01 Comparative Example 1 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:0. 3.96 Comparative Example 2 The molar ratio of acrylic acid monomer to dopamine hydrochloride is 1:1; the reaction solution in step (2) is allowed to stand for 120 hours. 2.34 From the test results in Table 1, it can be seen that the bonding strength increases with the increase in the proportion of dopamine hydrochloride. When the dopamine hydrochloride content reaches a certain proportion, the effect on the bonding strength is not particularly obvious; as the reaction solution is left standing for an increasing time, the bonding strength first increases and then decreases. At the beginning of the increase in oxidation time, there is a lot of dopamine that is not oxidized, and forms immediate adhesion with the tissue surface through non-covalent action, while dopamine quinone slowly forms covalent bonds with amine and thiol groups to provide long-term adhesion. As the oxidation time becomes longer, dopamine is oxidized into dopamine quinone, and the immediate adhesion formed by the dopamine group and the tissue surface decreases. As more tissue fluid fills the space between the patch and the tissue, under wet conditions, the formation of hydrogen bonds and chemical bonds within the dopamine quinone molecule is accelerated, resulting in a decrease in bonding strength.

[0036] (2) Evaluation of antibacterial properties The antibacterial properties of the tissue patches of each embodiment and comparative example were tested by the inhibition zone method. After ultraviolet irradiation sterilization on a clean bench, 100 μL of Escherichia coli and Staphylococcus aureus suspension (1*10 8 CFU / mL) onto LB medium and spread it evenly using a spreading rod to ensure that the colonies completely cover the plate counting agar. Then stick a tissue patch with a diameter of 10 mm on the medium and culture it in a 37°C biochemical incubator for 24 h. After 24 h of culture, take out the medium to observe the bacterial growth and record the diameter of the inhibition zone.

[0037] Table 2 Results of the antibacterial zone diameters of tissue patches prepared in various embodiments and comparative examples As can be seen from Table 2, no inhibition zone appears in the tissue patch of comparative example 1, indicating that it has no antibacterial property. After the dopamine component is introduced, the inhibition zone begins to appear, and with the increase of dopamine and dopamine quinone, the inhibition zone becomes more and more obvious, and the diameter of the inhibition zone becomes larger and larger. It can be seen that the antibacterial tissue patch prepared in the present invention has good antibacterial property.

[0038] In addition, the present invention also uses ATR-FTIR to perform infrared testing on the antibacterial tissue patch prepared in Example 1, and its infrared absorption spectrum is as follows: Figure 1 The infrared characteristic absorption peaks corresponding to the antibacterial tissue adhesion layer are as follows: 2941.85cm -1 It is the characteristic absorption peak of acrylic acid CH; 1653.70cm -1 , 1730.47cm -1 It is the characteristic absorption peak of vinyl pyrrolidone C=O; 1642.57cm -1 It is the characteristic absorption peak of dopamine quinone C=O, 1550.51 cm -1 , 1495.38 cm -1 , 1476.24 cm -1 It is the characteristic absorption peak of benzene ring C=C.

[0039] In summary, the present invention has developed an antibacterial tissue patch with excellent adhesion, and its preparation method and application. By using a polymer material with catechol groups and quinone groups, a non-covalent interaction is formed with the tissue, and the quinone groups form covalent bonds with the amine groups on the tissue surface, which can not only adhere to the tissue surface immediately, but also provide long-term adhesion. At the same time, it has good biological activities such as antibacterial, bactericidal, and anti-tumor.

[0040] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with this technology to understand the contents of the present invention and implement them accordingly. They cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. An antibacterial tissue patch with excellent adhesion, characterized in that: The method is formed by alternately coating at least one carrier layer and at least one antibacterial tissue adhesion layer; the carrier layer is a film-forming polymer. The antibacterial tissue adhesion layer is an antibacterial copolymer, and the antibacterial copolymer is poly(N-vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopamine quinone acrylate).

2. The antibacterial tissue patch with excellent adhesion according to claim 1, characterized in that: The film-forming polymer is a mixture of one or more selected from polylactic acid, polyglycolic acid, polycaprolactone and polyhydroxyalkanoate.

3. An antibacterial tissue patch with excellent adhesion according to claim 1, characterized in that: The antibacterial copolymer is formed by copolymerizing dopamine, acrylic acid and N-vinyl pyrrolidone and then inducing partial oxidation. The molar ratio of dopamine, acrylic acid and N-vinyl pyrrolidone is 0.6-1.4:1:

1.

4. A method for preparing an antibacterial tissue patch with excellent adhesion according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) dissolving the film-forming polymer, coating it, and drying it to obtain a carrier layer; (2) Under a nitrogen atmosphere, acrylic acid, dopamine hydrochloride and a condensing agent are subjected to condensation esterification to prepare dopamine acrylate; (3) adding N-vinyl pyrrolidone and a photoinitiator to the dopamine acrylate, stirring evenly under light-proof conditions, and then polymerizing under ultraviolet light, and then standing for purification to obtain poly(N-vinyl pyrrolidone-acrylic acid-dopamine acrylate-dopaminequinone acrylate); (4) The poly(N-vinylpyrrolidone-acrylic acid-dopamine acrylate-dopaminequinone acrylate) is dissolved, and then coated on the carrier layer and dried, and the coating and drying processes are repeated to obtain the antibacterial tissue patch with excellent adhesion.

5. The method for preparing an antibacterial tissue patch with excellent adhesion according to claim 4, characterized in that: The molar ratio of the acrylic acid to the condensing agent in step (2) is 1:

2.

6. The method for preparing an antibacterial tissue patch with excellent adhesion according to claim 4, characterized in that: The condensing agent in step (2) is a mixed solution of EDC and NHS, and the molar ratio of the EDC to the NHS is 1:

1.

7. The method for preparing an antibacterial tissue patch with excellent adhesion according to claim 4, characterized in that: The reaction temperature in step (2) is room temperature, and the reaction time is 6 to 18 hours.

8. The method for preparing an antibacterial tissue patch with excellent adhesion according to claim 4, characterized in that: In step (3), the photoinitiator is a mixture of one or more selected from TPO, MBF and BP, and the molar ratio of the dopamine acrylate, the N-vinyl pyrrolidone and the photoinitiator is 1:1:0.

04.

9. The method for preparing an antibacterial tissue patch with excellent adhesion according to claim 4, characterized in that: The UV polymerization time in step (3) is 10 to 30 minutes, and the standing time is 24 to 72 hours.

10. Use of the antibacterial tissue patch with excellent adhesion according to any one of claims 1 to 3 in a functional composite material.

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