Tissue adhesive and its preparation method and application

Abstract

The present invention relates to the field of medical material technology, and specifically provides a method for preparing a tissue adhesive. The preparation method of the present invention comprises: (1) mixing an amino acid raw material, acrylic acid, a cross-linking agent, a solvent and an initiator, and obtaining a tissue adhesive precursor through a polymerization reaction; (2) mixing the tissue adhesive precursor and a polybasic amino acid to obtain a tissue adhesive; the amino acid raw material comprises an acidic amino acid and / or a polyacidic amino acid; and the cross-linking agent is prepared by reacting a glycidyl acrylate compound and a polyacidic amino acid. The present invention uses a double-bond modified polyacidic amino acid as a macromolecular cross-linking agent, and realizes a stable covalent single network structure through free radical copolymerization with an acrylic acid monomer, and then forms hydrogen bonds and electrostatic interactions with the polybasic amino acid to obtain a reinforced and toughened non-covalent network, thereby realizing a double-network reinforced tissue adhesive network structure, which effectively improves the anti-swelling performance of the tissue adhesive.

Description

Technical Field

[0001] The present invention relates to the technical field of medical materials, and more particularly to a tissue adhesive and a preparation method and application thereof. Background Art

[0002] Every year, tens of millions of patients suffer from various tissue traumas due to accidents, chronic wounds (such as diabetic ulcers), and surgical procedures. With the advancement of modern medical technology, people have increasingly demanding requirements for postoperative recovery. Tissue adhesives can provide mechanical support for tissues while sealing wounds and preventing leakage, thus avoiding additional trauma and leakage caused by sutures and staples, as well as serious mechanical property mismatches between tissues and fixation devices.

[0003] Tissue adhesives can be used for closed hemostasis, skin tissue regeneration, wound closure and postoperative repair. They have the advantages of good hemostasis, avoiding leakage of liquids and air, and avoiding secondary injuries caused by needle punctures. They are easy to use and greatly reduce patient pain and operation time. Tissue adhesives must not only have sufficient adhesion properties, but also have the flexibility to conform to the underlying tissue and the ability to maintain these properties under humid conditions, as well as a low swelling index to avoid compressing nerves and blood vessels. Ideally, tissue adhesives should also be easy to produce, store and use. Tissue adhesives are usually used in the presence of water and body fluids (such as blood and sweat), so they must be able to achieve strong adhesion under humid conditions.

[0004] However, adhesion in wet conditions remains a significant challenge due to the interfacial hydration layer. Furthermore, the water absorption of tissue adhesives also poses a potential risk of tissue compression. Therefore, it is imperative to research and prepare wet tissue adhesives with anti-swelling properties. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a tissue adhesive and its preparation method and application. The tissue adhesive of the present invention has a covalent and non-covalent double cross-linked network structure, which effectively improves the anti-swelling performance of the tissue adhesive.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for preparing a tissue adhesive, comprising:

[0008] (1) mixing amino acid raw materials, acrylic acid, a cross-linking agent, a solvent, and an initiator to obtain a tissue glue precursor through a polymerization reaction;

[0009] (2) mixing a tissue glue precursor and polybasic amino acid to obtain a tissue adhesive;

[0010] The amino acid raw material includes acidic amino acids and / or polyacidic amino acids;

[0011] The cross-linking agent is prepared by reacting glycidyl acrylate compounds and polyacidic amino acids.

[0012] In the present invention, the step (1) is specifically as follows: mixing the polyacidic amino acid, acrylic acid, a crosslinking agent (polyacidic amino acid containing double bonds), a solvent and an initiator, and obtaining a tissue glue precursor through a polymerization reaction; the polymerization reaction time is 5 to 15 minutes, preferably 5 minutes.

[0013] Polybasic amino acids refer to macromolecules obtained by polymerizing basic amino acids (a few specific amino acids can be called basic amino acids, for example, lysine). Polyacidic amino acids refer to macromolecules obtained by polymerizing acidic amino acids.

[0014] In the present invention, the tissue glue precursor and the polybasic amino acid are mixed for 24 to 36 hours, preferably 24 hours.

[0015] In the present invention, the preparation method of the cross-linking agent includes: modifying the side groups of polyacidic amino acids with double bonds using a glycidyl acrylate compound; specifically, mixing the polyacidic amino acid and the glycidyl acrylate compound, adjusting the pH to 4.5-5, and reacting at 60-80°C for 12-24 hours to obtain the cross-linking agent; the reaction is preferably carried out under stirring conditions, and the stirring speed is 200-220 r / min; the glycidyl acrylate compound includes glycidyl methacrylate and / or glycidyl acrylate.

[0016] In the present invention, the molar ratio of the polyacidic amino acid to the glycidyl acrylate compound is (1-2):2, preferably 1:2; the polyacidic amino acid is mixed with the glycidyl acrylate compound in the form of a solution; the concentration of the polyacidic amino acid solution (w / v, g / 100 ml) is 10% to 15%.

[0017] In the present invention, the preparation method of the cross-linking agent further comprises: dialyzing the reaction solution in water for 5 days, changing the water every 12 hours; and freeze-drying the solution after the dialysis to obtain a double-bond modified polyacidic amino acid.

[0018] In the present invention, the mass ratio of the initiator, polyacidic amino acid, acrylic acid and cross-linking agent is 1:(10-70):(40-300):(2-20).

[0019] In one embodiment of the present invention, the mass ratio of the initiator, polyacidic amino acid, acrylic acid and cross-linking agent is 1:50:200:10.

[0020] In one embodiment of the present invention, the mass ratio of the initiator, polyacidic amino acid, acrylic acid and cross-linking agent is 1:30:125:5.

[0021] In one embodiment of the present invention, the mass ratio of the initiator, polyacidic amino acid, acrylic acid and cross-linking agent is 1:14:60:4.

[0022] In the present invention, the polymerization is initiated by ultraviolet light; the ultraviolet light has a wavelength of 365 nm and a power of 200 mW / cm 2 .

[0023] In the present invention, the solvent is preferably water, more preferably deionized water; the mass ratio of the initiator to the solvent is 1:(100-600), preferably 1:300 or 1:120 or 1:500.

[0024] In the present invention, the mass ratio of the tissue glue precursor to the polybasic amino acid is 1:(0.01-10), preferably 5:3.

[0025] In the present invention, the polybasic amino acid is mixed with the tissue glue precursor in the form of a solution; more preferably, the tissue glue precursor is immersed in the polybasic amino acid solution;

[0026] The concentration (w / v) of the polybasic amino acid solution is 15% to 20%, preferably 15% to 17%.

[0027] In the present invention, the initiator is preferably a photoinitiator; in one embodiment of the present invention, the initiator is preferably photoinitiator L2959.

[0028] In the present invention, the molecular weight of the polyacidic amino acid is 1100-1500 KDa, preferably 1100 KDa;

[0029] The molecular weight of the polybasic amino acid is 3000-6000 Da, preferably 3500-5000 Da.

[0030] In the present invention, the polyacidic amino acid includes polyglutamic acid and / or polyaspartic acid, preferably polyglutamic acid;

[0031] The polybasic amino acid includes one or more of polylysine, polyarginine, and polyhistidine, preferably polylysine.

[0032] The present invention also provides a tissue adhesive prepared by the preparation method of the tissue adhesive.

[0033] The present invention also provides the tissue adhesive prepared by the preparation method of the tissue adhesive, or the use of the tissue adhesive in preparing biomaterials.

[0034] Compared with other methods, the present invention has the following beneficial technical effects:

[0035] (1) The present invention uses double-bond-modified polyacidic amino acids as macromolecular crosslinkers, which, through free radical copolymerization with acrylic monomers, achieve a stable covalent single network structure. This structure then forms hydrogen bonds and electrostatic interactions with polybasic amino acids to form a reinforced and toughened non-covalent network, thereby achieving a dual-network-reinforced tissue adhesive network structure. The tissue adhesive of the present invention has a covalent and non-covalent dual crosslinked network structure, which effectively improves the anti-swelling properties of the tissue adhesive.

[0036] (2) The tissue adhesive of the present invention has electrostatic interaction, which greatly improves the wet adhesion stability of the tissue adhesive and can achieve long-term stable adhesion in an aqueous environment.

[0037] (3) The double-bond modified polyacidic amino acid used in the present invention is a macromolecular cross-linking agent with degradable properties and excellent biocompatibility.

[0038] (4) The technical solution of the present invention has excellent antibacterial properties and can effectively prevent tissue infection. DETAILED DESCRIPTION

[0039] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0040] To further illustrate the present invention, the following examples are provided for detailed description. The raw materials used in the following examples are all commercially available products. The photoinitiator L2959 used in the examples is primarily composed of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone; polyglutamic acid (Mw = 1100 kDa) and polylysine (Mw = 3500-5000 Da) purchased from Nanjing Xuankai Biotechnology Co., Ltd.

[0041] Example 1

[0042] (1) preparing double-bond modified polyglutamic acid, specifically comprising the following steps:

[0043] (a) Dissolve polyglutamic acid in deionized water to a concentration of 10% w / v;

[0044] (b) adding glycidyl methacrylate to the polyglutamic acid solution, and adjusting the pH of the mixed solution to 4.5, wherein the molar ratio of glycidyl methacrylate to polyglutamic acid is 1:2;

[0045] (c) stirring at 60° C. and 200 rpm for 12 h to obtain a reaction solution;

[0046] (d) The reaction solution was dialyzed in deionized water for 5 days, with the water changed every 12 h;

[0047] (e) After dialysis, the solution is freeze-dried to obtain double-bond modified polyglutamic acid.

[0048] (2) Adding polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 to deionized water to obtain a mixed solution, wherein the ratio of polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 is as follows: by weight, 5 parts of polyglutamic acid, 20 parts of acrylic acid, 1 part of double-bond modified polyglutamic acid, 0.1 part of photoinitiator L2959 and 50 parts of deionized water are weighed respectively.

[0049] (3) Spreading the mixed solution obtained in step (2) in a mold, and initiating polymerization with ultraviolet light to obtain a tissue glue precursor, wherein the ultraviolet light wavelength is 365nm and the power is 200mw / cm 2 , the irradiation time is 5min.

[0050] (4) 5 g of tissue glue precursor was soaked in 20 ml of polylysine solution (15% w / v, polylysine content was 3 g) for 24 h to obtain a wet tissue adhesive with anti-swelling properties.

[0051] Example 2

[0052] (1) preparing double-bond modified polyglutamic acid, specifically comprising the following steps:

[0053] (a) Dissolve polyglutamic acid in deionized water at a concentration of 12% w / v;

[0054] (b) adding glycidyl methacrylate and adjusting the pH of the mixed solution to 4.5, wherein the molar ratio of glycidyl methacrylate to polyglutamic acid is 1:2;

[0055] (c) stirring at 60° C. and 200 rpm for 12 h to obtain a reaction solution;

[0056] (d) The solution was dialyzed in deionized water for 5 days, with the water changed every 12 h;

[0057] (e) After dialysis, the solution is freeze-dried to obtain double-bond modified polyglutamic acid.

[0058] (2) Adding polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 to deionized water to obtain a mixed solution, wherein the ratio of polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 is as follows: 6 parts of polyglutamic acid, 25 parts of acrylic acid, 1 part of double-bond modified polyglutamic acid, 0.2 parts of photoinitiator L2959 and 60 parts of deionized water are weighed respectively.

[0059] (3) Spreading the mixed solution obtained in step (2) in a mold, and initiating polymerization with ultraviolet light to obtain a tissue glue precursor, wherein the ultraviolet light wavelength is 365nm and the power is 200mw / cm 2 , the irradiation time is 5min.

[0060] (4) 5 g of tissue glue precursor was soaked in 20 ml of polylysine solution (17% w / v, polylysine content was 3.4 g) for 24 h to obtain a wet tissue adhesive with anti-swelling properties.

[0061] Example 3

[0062] (1) preparing double-bond modified polyglutamic acid, specifically comprising the following steps:

[0063] (a) Dissolve polyglutamic acid in deionized water to a concentration of 15% w / v;

[0064] (b) adding glycidyl methacrylate and adjusting the pH of the mixed solution to 4.5, wherein the molar ratio of glycidyl methacrylate to polyglutamic acid is 1:2;

[0065] (c) stirring at 60° C. and 200 rpm for 12 h to obtain a reaction solution;

[0066] (d) The solution was dialyzed in deionized water for 5 days, with the water changed every 12 h;

[0067] (e) After dialysis, the solution is freeze-dried to obtain double-bond modified polyglutamic acid.

[0068] (2) Adding polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 to deionized water to obtain a mixed solution, wherein the ratio of polyglutamic acid, acrylic acid, double-bond modified polyglutamic acid and photoinitiator L2959 is as follows: by weight, weigh 7 parts of polyglutamic acid, 30 parts of acrylic acid, 2 parts of double-bond modified polyglutamic acid, 0.5 parts of photoinitiator L2959 and 60 parts of deionized water respectively.

[0069] (3) Spreading the mixed solution obtained in step (2) in a mold, and initiating polymerization with ultraviolet light to obtain a tissue glue precursor, wherein the ultraviolet light wavelength is 365nm and the power is 200mw / cm 2 , the irradiation time is 5min.

[0070] (4) 5 g of tissue glue precursor was soaked in 20 ml of polylysine solution (17% w / v, polylysine content was 3.4 g) for 24 h to obtain a wet tissue adhesive with anti-swelling properties.

[0071] Test Example 1

[0072] The performance tests of Examples 1, 2 and 3 prepared by the present invention were carried out using the following method:

[0073] 1) Bond strength test:

[0074] The tissue adhesive obtained in the example was applied between two overlapping wet pig skins with an overlapping area of ​​1*1 cm 2 Gently press with a force of about 1 kPa for 5 seconds. Use a universal tensile testing machine to stretch at a speed of 50 mm / min to test the bond strength.

[0075] 2) Volume swelling rate test:

[0076] The obtained tissue adhesive was placed in deionized water and weighed every 12 hours until it stabilized. The volume swelling rate was calculated as follows:

[0077] SR(%)=(Wb-Wa) / Wa

[0078] Where SR represents the volume swelling ratio (%), Wb is the mass of tissue glue after swelling stabilization, and Wa is the initial mass before swelling.

[0079] The specific test results are shown in Table 1.

[0080] Table 1 Performance test table

[0081] Performance Characterization Example 1 Example 2 Example 3 Bond strength (kPa) 150.21 165.35 164.02 Volume swelling rate (%) 120.58 115.08 118.25

[0082] As can be seen from Table 1, the wet tissue adhesives prepared in Examples 1-3 of the present invention have excellent tissue adhesion and anti-swelling properties.

[0083] The above description of the disclosed embodiments will enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is to be construed in the widest manner consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a tissue adhesive, characterized in that: include: (1) Mixing polyacidic amino acid, acrylic acid, a cross-linking agent, a solvent and an initiator to obtain a tissue glue precursor through a polymerization reaction; (2) mixing the tissue glue precursor and the polybasic amino acid to obtain a tissue adhesive; The cross-linking agent is prepared by reacting glycidyl acrylate compounds and polyacidic amino acids; The preparation method of the cross-linking agent comprises: The polyacidic amino acid and glycidyl acrylate compound are mixed, the pH is adjusted to 4.5-5, and the mixture is reacted at 60-80°C for 12-24 hours to obtain a cross-linking agent; The glycidyl acrylate compound includes glycidyl methacrylate and / or glycidyl acrylate; The mass ratio of the initiator, polyacidic amino acid, acrylic acid and cross-linking agent is 1: (10-70): (40-300): (2-20); The mass ratio of the tissue glue precursor to the polybasic amino acid is 1: (0.01-10); The mass ratio of the initiator to the solvent is 1:(100~600).

2. The method for preparing the tissue adhesive according to claim 1, wherein: The polymerization reaction time is 5 to 15 min; The tissue glue precursor and the polybasic amino acid are mixed for 24 to 36 hours.

3. The method for preparing the tissue adhesive according to claim 1, wherein: The molecular weight of the polyacidic amino acid is 1100-1500 KDa; The molecular weight of the polybasic amino acid is 3000-6000 Da.

4. The method for preparing the tissue adhesive according to claim 1, wherein: The polymerization is ultraviolet light initiated polymerization.

5. The method for preparing the tissue adhesive according to claim 1, wherein: The polyacidic amino acid comprises polyglutamic acid and / or polyaspartic acid; The polybasic amino acid includes one or more of polylysine, polyarginine and polyhistidine.

6. The tissue adhesive prepared by the method for preparing the tissue adhesive according to any one of claims 1 to 5.

7. Use of the tissue adhesive according to claim 6 in preparing biomaterials.