Biological glue as well as preparation method and application thereof

Abstract

The invention relates to biological glue as well as a preparation method and application thereof. The biological glue comprises the following components: tannic acid, gelatin and polyvinylpyrrolidone, the polyvinylpyrrolidone comprises a mixture of polyvinylpyrrolidone K30, polyvinylpyrrolidone K60 and polyvinylpyrrolidone K90, and the polyvinylpyrrolidone K30 is a mixture of polyvinylpyrrolidone K60 and polyvinylpyrrolidone K90. The biological glue has good biocompatibility and high-strength adhesiveness.

Description

Technical Field

[0001] The present application relates to the field of biomedical materials, and in particular to a bioglue and a preparation method and application thereof. Background Art

[0002] In surgical procedures, medical bioadhesives are expected to become an ideal substitute for clinical sutures due to their strong adhesion, biocompatibility, and ease of operation. However, the current mainstream commercial tissue adhesives have some shortcomings. For example, cyanoacrylate (super glue), as a commercial adhesive with excellent adhesion properties, has potential biotoxicity. Fibrin-based adhesives and polyethylene glycol-based adhesives, such as COSEAL, can form a brittle matrix that is prone to cohesion and failure. The strong adhesion of adhesives mainly comes from the synergistic effect of the matrix: the overall adhesion of the matrix and the interfacial adhesion between the matrix and the biological tissue. The mechanical strength of the adhesive matrix usually affects the adhesion of the body. In addition, poor mechanical strength can lead to the failure of the adhesive matrix, while adhesives with low interfacial adhesion will break at the adhesion boundary (interfacial failure). In recent years, polymer-based adhesives have good biocompatibility, but poor mechanical properties and adhesion properties have hindered their application. Although relevant research has been devoted to improving the above problems, such as enhancing adhesion through cross-linking networks or interfacial interactions, it is still very challenging to simultaneously improve the mechanical properties and adhesion properties of adhesives. For example, a bioabsorbable composite hemostatic material and its preparation method and application are prepared from the following raw material components: polyvinyl alcohol, gelatin and tannic acid, wherein the molecular weight of the polyvinyl alcohol is in the range of 10,000 to 50,000 Da, and the weight ratio of the polyvinyl alcohol, gelatin and tannic acid is 0.2 to 0.4:1 to 1.5:1. However, the bonding strength is still relatively low.

[0003] Therefore, it is imperative to develop a synthetic medical bioglue with good biocompatibility and high-strength adhesive properties. Summary of the invention

[0004] Based on this, one embodiment of the present application provides a bioglue and a preparation method thereof, wherein the bioglue has good biocompatibility and high-strength adhesion.

[0005] The technical solution includes:

[0006] Bio-glue, comprising the following components: tannic acid, gelatin and polyvinyl pyrrolidone;

[0007] The polyvinyl pyrrolidone includes a mixture of polyvinyl pyrrolidone K30, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K90.

[0008] In one embodiment, the mass ratio of the polyvinyl pyrrolidone K90, the polyvinyl pyrrolidone K60 and the polyvinyl pyrrolidone K30 is (4-7):(2-3):(1-3).

[0009] In one embodiment, the mass ratio of the gelatin to the polyvinyl pyrrolidone is 1:(1-8), and the mass ratio of the tannic acid to the polyvinyl pyrrolidone is (1-3):1.

[0010] The method for preparing the bio-glue comprises the following steps:

[0011] The aqueous solution of polyvinyl pyrrolidone is mixed with the aqueous solution of gelatin to prepare solution A; and the aqueous solution of tannic acid is mixed with solution A to prepare bio-glue.

[0012] In one embodiment, the polyvinyl pyrrolidone is mixed with water at a temperature of 60° C.-80° C. and a speed of 500 rpm-1000 rpm to prepare the aqueous solution of polyvinyl pyrrolidone.

[0013] Optionally, in the aqueous solution of polyvinyl pyrrolidone, the concentration of polyvinyl pyrrolidone is 10% w / v-30% w / v.

[0014] In one embodiment, the gelatin is mixed with water at a temperature of 55°C-65°C to prepare an aqueous solution of the gelatin.

[0015] Optionally, the concentration of gelatin in the aqueous solution of gelatin is 2% w / v-10% w / v.

[0016] In one embodiment, the concentration of tannic acid in the aqueous solution of tannic acid is 10% w / v-30% w / v.

[0017] In one embodiment, the conditions for mixing the aqueous solution of tannic acid with the solution A include stirring at a rate of 500 rpm-1000 rpm for 18 h-24 h.

[0018] In one embodiment, after the aqueous solution of tannic acid is mixed with the solution A, the mixture is centrifuged at 8000 rpm-10000 rpm for 5 min-10 min, and the resulting precipitate is washed with a PBS solution and then concentrated.

[0019] The bioglue is used in preparing medical suture, injury repair or hemostasis materials.

[0020] Compared with the traditional technology, this application has the following beneficial effects:

[0021] 1. Excellent adhesion performance

[0022] (1) Hydrogen bond-mediated dynamic cross-linking network: The hydrogen bond interaction between the tannic acid in the bio-glue of the present application and the polyvinyl pyrrolidone mixed with different molecular weights forms a dynamic cross-linking network with better performance. This dynamic cross-linking network has a strong interaction with the tissue surface. This interaction not only enhances the adhesion of the adhesive, but also improves the stability and durability of the adhesion. In addition, a mechanical interlocking effect can be generated through the microscopic rough structure of the tissue surface. This mechanical interlocking effect forms a "lock"-like structure between the adhesive layer and the tissue surface, which is difficult to separate, further synergistically enhancing the adhesion strength of the bio-glue.

[0023] (2) Covalent bond-enhanced interfacial bonding: The tannic acid and gelatin in the bio-glue of the present application can further enhance the bonding force between the bio-glue and biological tissues through a variety of chemical covalent bonds (Schiff base reaction, etc.), thereby improving the durability and stability of adhesion.

[0024] 2. Good biocompatibility: Tannic acid and gelatin are both derived from natural substances, are safe, have good biocompatibility, and reduce irritation and immune response to biological tissues. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0026] Figure 1 Schematic diagram of the working principle of components in bioglue. DETAILED DESCRIPTION

[0027] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below. In the following description, many specific details are set forth to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present application, so the present application is not limited by the specific embodiments disclosed below.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0029] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0030] In this article, "optionally", "optional", and "optional" mean optional, that is, any one of the two parallel solutions of "yes" or "no". If there are multiple "optional" in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent. Unless otherwise specified, the descriptions of "optionally include", "optionally include", etc. in this application, taking "optionally include" as an example, mean "may include or not include".

[0031] Herein, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers in the numerical interval, including the two endpoint integers of the numerical range, and each integer between the two endpoints, is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows broadly including numerical interval types such as percentage intervals, ratio intervals, and ratio intervals.

[0032] At present, various types of medical biological glue, such as fibrin adhesive, BioGlue, cyanoacrylate adhesive, GRF adhesive, polyethylene glycol adhesive, etc., have been used in clinical practice, but they have certain limitations. Although fibrin adhesive has been used for a long time and has a wide range of applications, it is gradually being replaced due to its relatively low bonding strength; semi-synthetic soft tissue adhesives such as BioGlue and GRF adhesives, although the bonding strength meets the use requirements, the aldehyde substances used in the products have potential toxic effects on the human body, so the dosage must be considered when using them. Since the raw materials of various synthetic medical biological glues are gradually replaced by recognized non-toxic raw materials, it is imperative to develop a synthetic, non-toxic medical biological glue with high-strength bonding properties.

[0033] Tannic acid (TA) is a natural polyphenol with pyrogallol groups, which is similar to the catechol groups of mussel protein (dopamine). However, the number of phenolic -OH groups (25 units per TA molecule) in TA is higher than that of dopamine molecules, which enhances the adhesion of tannic acid to almost all inorganic and organic surfaces and improves the mechanical strength. Therefore, TA is considered to be a versatile strong molecular adhesive that can form multi-tooth interactions with polymer chains, resulting in a stable boundary layer between the adhesive and different substrates. Adhesion mediated by dynamic non-covalent interactions (such as hydrogen bonds, hydrophobic interactions, and cation-π interactions) and covalent bonds (such as coordination bonds, Schiff base reactions, and Michael addition reactions, etc.) is used to optimize the above problems for medical bioadhesives to achieve better hemostasis and achieve the best sealing results in clinical applications.

[0034] An embodiment of the present application provides a bio-glue, which includes the following components: tannic acid, gelatin and polyvinyl pyrrolidone of different molecular weights.

[0035] In a preferred specific example, the polyvinyl pyrrolidone with different molecular weights includes a mixture of polyvinyl pyrrolidone K30, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K90.

[0036] Polyvinylpyrrolidone, PVP for short, is a synthetic water-soluble polymer compound. PVP is divided into several grades according to the average molecular weight, and is generally classified by the K value of the Fikentscher method. Its commercial K value is between K15 and K120.

[0037] The present application mainly contains bioadhesive (or bioadhesive) of tannic acid / polyvinyl pyrrolidone of different molecular weights / gelatin ternary complex; wherein tannic acid (TA) can provide cohesive strength for macromolecular networks such as polyvinyl pyrrolidone (PVP); the covalent bond of tannic acid-gelatin improves the mechanical properties of the material. The bioadhesive has good biocompatibility and excellent strong adhesion, and can effectively adhere to different types of tissues to meet different medical requirements such as wound sealing, tissue repair, and liquid blocking.

[0038] Compared with the glue material formed by tannic acid / gelatin / polyvinyl alcohol or tannic acid / gelatin / Pluronic, the biological glue of the present application has better adhesion.

[0039] In a specific example, the bioglue has an interpenetrating network structure.

[0040] Interpenetrating network (IPN) hydrogel is a three-dimensional network structure formed by two or more different polymer networks interpenetrating and entangled with each other through physical or chemical means. This structure gives IPN hydrogel unique performance characteristics and application potential.

[0041] In this application, if Figure 1 As shown in Figure 1, on the one hand, tannic acid contains multiple hydroxyl (-OH) groups, while polyvinyl pyrrolidone contains a large number of carbonyl groups (C=O). Since the hydrogen atoms of the hydroxyl groups can form hydrogen bond interactions with the oxygen atoms of the carbonyl groups, this interaction establishes a dynamic cross-linking network between tannic acid and polyvinyl pyrrolidone; on the other hand, gelatin is a natural polymer material with multiple functional groups, such as amino groups and carboxyl groups. Therefore, the phenolic hydroxyl groups in tannic acid can react chemically with the functional groups in gelatin to form covalent bonds (Schiff base reaction, etc.). The researchers unexpectedly found that the use of a mixture of polyvinyl pyrrolidone with different molecular weights can further significantly enhance the adhesion properties of the entire interpenetrating network system. This may be because the mixture of polyvinyl pyrrolidone with different molecular weights (such as polyvinyl pyrrolidone K90, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K30) can directly affect the forces between polymer molecules based on their specific non-uniform molecular weights and molecular segments, thereby significantly improving the adhesion properties of the material.

[0042] In a specific example, the mass ratio of polyvinyl pyrrolidone K90, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K30 is (4-7):(2-3):(1-3).

[0043] In a specific example, the mass ratio of polyvinyl pyrrolidone K90, the polyvinyl pyrrolidone K60 and the polyvinyl pyrrolidone K30 may be (5-7):(2-3):(1-2).

[0044] In a specific example, the mass ratio of polyvinyl pyrrolidone K90, the polyvinyl pyrrolidone K60 and the polyvinyl pyrrolidone K30 can be selected to be 7:2:1.

[0045] In a specific example, the mass ratio of gelatin to polyvinyl pyrrolidone is 1:(1-8). In a specific example, the mass ratio of gelatin to polyvinyl pyrrolidone can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, or a range between any two of the above ratios.

[0046] In a specific example, the mass ratio of tannic acid to polyvinyl pyrrolidone is (1-3): 1. In a specific example, the mass ratio of tannic acid to polyvinyl pyrrolidone can be 1:1, 2:1, 3:1, or a range between any two of the above ratios.

[0047] An embodiment of the present application provides a method for preparing the bio-glue, which has the advantages of simple process, good adhesion of the prepared bio-glue, etc., and includes the following steps S10 and S20:

[0048] Step S10: Mixing an aqueous solution of polyvinyl pyrrolidone with an aqueous solution of gelatin to prepare solution A.

[0049] In a specific example, polyvinyl pyrrolidone of different molecular weights is mixed with water, and an aqueous solution of polyvinyl pyrrolidone is prepared at a temperature of 60°C-80°C and a rate of 500rpm-1000rpm. Optionally, the temperature is 60°C, 65°C, 70°C, 75°C or 80°C, and optionally, the rate is 500rpm, 600rpm, 700rpm, 800rpm, 900rpm or 1000rpm.

[0050] In a specific example, the concentration of polyvinyl pyrrolidone in the aqueous solution of polyvinyl pyrrolidone is 10% w / v-30% w / v. In a specific example, the concentration of polyvinyl pyrrolidone in the aqueous solution of polyvinyl pyrrolidone can be selected as 10% w / v, 15% w / v, 20% w / v, 25% w / v, 30% w / v, or a range between any two of the above values.

[0051] In a specific example, gelatin is mixed with water to prepare an aqueous solution of gelatin at a temperature of 55°C-65°C, optionally, the temperature is 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C or 65°C.

[0052] In a specific example, the concentration of gelatin in the aqueous solution of gelatin is 2% w / v-10% w / v. In a specific example, the concentration of gelatin in the aqueous solution of gelatin can be 2% w / v, 2.5% w / v, 3% w / v, 4% w / v, 5% w / v, 6% w / v, 7% w / v, 8% w / v, 9% w / v, 10% w / v, or a range between any two of the above values.

[0053] Step S20: Mixing the aqueous solution of tannic acid with solution A to prepare bio-glue.

[0054] In a specific example, tannic acid is mixed with water and stirred at room temperature to prepare an aqueous solution of tannic acid.

[0055] In a specific example, the concentration of tannic acid in the aqueous solution of tannic acid is 10% w / v-30% w / v. In a specific example, the concentration of tannic acid in the aqueous solution of tannic acid can be selected as 10% w / v, 15% w / v, 20% w / v, 25% w / v, 30% w / v, or a range between any two of the above ratios.

[0056] In a specific example, the conditions for mixing the aqueous solution of tannic acid with solution A include stirring overnight at a rate of 500-1000 rpm, optionally, at a rate of 500 rpm, 600 rpm, 700 rpm, 800 rpm, 900 rpm or 1000 rpm.

[0057] In a specific example, after the aqueous solution of tannic acid is mixed with solution A, the mixture is centrifuged at 8000 rpm to 10000 rpm for 5 to 10 min, and the resulting precipitate is washed with a PBS solution and then concentrated, and can be optionally stored at 4°C in the dark for later use.

[0058] In a specific example, the pH value of the PBS solution is 6-7.4.

[0059] One embodiment of the present application provides the use of the bioglue in preparing medical suture, injury repair or hemostatic materials.

[0060] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0061] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0062] Example 1

[0063] This embodiment provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0064] The preparation method of the medical biological glue comprises the following steps:

[0065] (1) 10 g of polyvinyl pyrrolidone (K90:K60:K30 mass ratio of 5:3:2) was added to 50 mL of deionized water and stirred at 500 rpm in a water bath at 80 °C to completely dissolve the polyvinyl pyrrolidone to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0066] (2) Add 2.5 g of gelatin to 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 2.5 w / v% gelatin solution.

[0067] (3) Add 10 g of tannic acid into 50 mL of deionized water and stir at room temperature to obtain a 20 w / v% tannic acid solution.

[0068] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:8.

[0069] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0070] Example 2

[0071] This embodiment provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0072] The preparation method of the medical biological glue comprises the following steps:

[0073] (1) 10 g of polyvinyl pyrrolidone (K90:K60:K30 mass ratio 7:2:1) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the polyvinyl pyrrolidone to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0074] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0075] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0076] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0077] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0078] Example 3

[0079] This embodiment provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0080] The preparation method of the medical biological glue comprises the following steps:

[0081] (1) 10 g of polyvinyl pyrrolidone (K90:K60:K30 mass ratio 4:3:3) was added to 50 mL of deionized water and stirred at 500 rpm in a water bath at 80 °C to completely dissolve the polyvinyl pyrrolidone to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0082] (2) Add 2.5 g of gelatin to 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 2.5 w / v% gelatin solution.

[0083] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0084] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:8.

[0085] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0086] Example 4

[0087] The difference from Example 2 is that the preparation method of the medical biological glue comprises the following steps:

[0088] (1) 15 g of polyvinyl pyrrolidone (K90:K60:K30 mass ratio 7:2:1) was added to 50 mL of deionized water and stirred at 750 rpm in a 60 °C water bath to completely dissolve the mixture to prepare a 30 w / v% polyvinyl pyrrolidone solution.

[0089] (2) Add 10 g of gelatin into 100 mL of deionized water and heat in a 55 °C water bath to completely dissolve it to obtain a 10 w / v% gelatin solution.

[0090] (3) Add 15 g of tannic acid to 50 mL of deionized water to obtain a 30 w / v% tannic acid solution at room temperature.

[0091] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:3.

[0092] (5) 10 mL of tannic acid solution was added to the homogenized precursor solution and stirred evenly at 750 rpm overnight for 20 h. Then, the mixture was centrifuged at 9 000 rpm for 8 min, fully concentrated with PBS pH=6 solution, and finally stored at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone was 1:1.

[0093] Example 5

[0094] The difference from Example 1 is that the preparation method of the medical biological glue comprises the following steps:

[0095] (1) 5 g of polyvinyl pyrrolidone (K90:K60:K30 mass ratio 7:2:1) was added to 50 mL of deionized water and stirred at 1000 rpm in a 60 °C water bath to completely dissolve the polyvinyl pyrrolidone to prepare a 10 w / v% polyvinyl pyrrolidone solution.

[0096] (2) Add 5 g of gelatin to 50 mL of deionized water and heat in a 65°C water bath to completely dissolve it to obtain a 10 w / v% gelatin solution.

[0097] (3) Add 5 g of tannic acid to 50 mL of deionized water and stir at room temperature to obtain a 10 w / v% tannic acid solution.

[0098] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin solution to polyvinyl pyrrolidone is 1:1.

[0099] (5) 10 mL of tannic acid solution was added to the homogenized precursor solution and stirred evenly at 1000 rpm overnight for 24 h. Then, the mixture was centrifuged at 8 000 rpm for 10 min, fully concentrated with PBS pH = 7.4 solution, and finally stored at 4 ° C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone was 1:1.

[0100] Comparative Example 1

[0101] Comparative Example 1 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0102] The method for preparing the medical bio-glue comprises the following steps:

[0103] (1) 10 g of polyvinyl pyrrolidone (K90:K30 mass ratio 7:3) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0104] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0105] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0106] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0107] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0108] Comparative Example 2

[0109] Comparative Example 2 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0110] The preparation method of the medical biological glue comprises the following steps:

[0111] (1) 10 g of polyvinyl pyrrolidone (K90:K60 mass ratio 7:3) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0112] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0113] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0114] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0115] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0116] Comparative Example 3

[0117] Comparative Example 3 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0118] The preparation method of the medical biological glue comprises the following steps:

[0119] (1) 10 g of polyvinyl pyrrolidone (K90:K30 mass ratio 6:4) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0120] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0121] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0122] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0123] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0124] Comparative Example 4

[0125] Comparative Example 4 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0126] The preparation method of the medical biological glue comprises the following steps:

[0127] (1) 10 g of polyvinyl pyrrolidone (K90:K60 mass ratio 6:4) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0128] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0129] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0130] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0131] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0132] Comparative Example 5

[0133] Comparative Example 5 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0134] The preparation method of the medical biological glue comprises the following steps:

[0135] (1) 10 g of polyvinyl pyrrolidone (K90) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0136] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0137] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0138] (4) Slowly add 10 mL of gelatin solution into 10 mL of polyvinyl pyrrolidone solution, and stir continuously at 500 rpm to obtain a homogeneous precursor solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone is 1:4.

[0139] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0140] Comparative Example 6

[0141] Comparative Example 6 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0142] The preparation method of the medical biological glue comprises the following steps:

[0143] (1) 10 g of polyvinyl pyrrolidone (K60) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution, thereby preparing a 20 w / v% polyvinyl pyrrolidone solution.

[0144] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0145] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0146] (4) 10 mL of gelatin solution was slowly added into 10 mL of polyvinyl pyrrolidone solution, and the mixture was continuously stirred at 500 rpm to obtain a solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone was 1:4.

[0147] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0148] Comparative Example 7

[0149] Comparative Example 7 provides a medical biological glue, the main components of which include tannic acid, polyvinyl pyrrolidone and gelatin.

[0150] The preparation method of the medical biological glue comprises the following steps:

[0151] (1) 10 g of polyvinyl pyrrolidone (K30) was added to 50 mL of deionized water and stirred continuously at 500 rpm in a water bath at 80 °C to completely dissolve the solution to prepare a 20 w / v% polyvinyl pyrrolidone solution.

[0152] (2) Add 5 g of gelatin into 100 mL of deionized water and heat in a 60 °C water bath to completely dissolve it to obtain a 5 w / v% gelatin solution.

[0153] (3) Add 10 g of tannic acid to 50 mL of deionized water to obtain a 20 w / v% tannic acid solution at room temperature.

[0154] (4) 10 mL of gelatin solution was slowly added into 10 mL of polyvinyl pyrrolidone solution, and the mixture was continuously stirred at 500 rpm to obtain a solution, wherein the mass ratio of gelatin to polyvinyl pyrrolidone was 1:4.

[0155] (5) Add 30 mL of tannic acid solution to the homogenized precursor solution and stir evenly at 500 rpm overnight for 18 h. Then, centrifuge the mixture at 10,000 rpm for 5 min, fully concentrate with PBS pH=6 solution, and finally store at 4 °C in the dark for later use. The mass ratio of tannic acid to polyvinyl pyrrolidone is 3:1.

[0156] Comparative Example 8

[0157] Comparative Example 8 provides a biomaterial mainly comprising tannic acid, polyvinyl alcohol and gelatin.

[0158] The method for preparing the biomaterial comprises the following steps:

[0159] (1) Add 3 g of polyvinyl alcohol (molecular weight: 30,000) and 10 g of gelatin powder into a flask containing 50 mL of deionized water, stir and dissolve at 90 °C for 1 h to obtain a uniform mixed solution;

[0160] (2) adding 10 g of tannic acid to the uniformly mixed solution of step (1), maintaining the temperature at 60° C. and stirring until a composite hemostatic material is obtained after self-assembly;

[0161] (3) The composite hemostatic material in step (2) is taken out, placed in a vacuum drying oven at 60° C. and dried for 24 h, then taken out and ground to obtain the target product.

[0162] test

[0163] Adhesion performance test of medical bioadhesive

[0164] Fresh pigskin tissue was selected to simulate human tissue. Fresh pigskin was purchased from the local market and cut into uniformly sized rectangles, with a size of about 4.0 cm × 1.5 cm. Excess fat on the pigskin tissue was removed, and the thickness should not exceed 5 mm. The cut pigskin was first washed with a 0.9% sodium chloride solution, and then soaked in a pH = 7.4 PBS buffer overnight to keep the pigskin moist. The pre-prepared medical bio-glue was evenly applied to the surface of a piece of pigskin, with an area of ​​about 1.5 cm × 1.5 cm, and then another piece of pigskin was covered on the upper layer of the medical bio-glue, and 10 N pressure was applied at the same time. The sample was fixed between two clamps, and the tensile rate was set to 1 mm / min. The bonded pigskin was taken out for tensile shear test until it was completely pulled apart. At the same time, the maximum tensile stress was recorded, and the adhesion strength of the medical bio-glue to the pigskin tissue was calculated. The average value was taken after 5 groups were tested in parallel. The test results are shown in Table 1-1 and Table 1-2. It can be seen that the maximum adhesion strength of Example 2 is 53.27 ± 1.81 kPa.

[0165] Table 1-1

[0166]

[0167] Table 1-2

[0168]

[0169] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are 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.

[0170] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the attached claims, and the description may be used to interpret the content of the claims.

Claims

1. Bioglue, characterized in that The biological glue comprises the following components: tannic acid, gelatin and polyvinyl pyrrolidone; The polyvinyl pyrrolidone includes a mixture of polyvinyl pyrrolidone K30, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K90.

2. The bioglue according to claim 1, characterized in that The mass ratio of the polyvinyl pyrrolidone K90, the polyvinyl pyrrolidone K60 and the polyvinyl pyrrolidone K30 is (4-7):(2-3):(1-3).

3. The bioglue according to claim 1 or 2, characterized in that The mass ratio of the gelatin to the polyvinyl pyrrolidone is 1:(1-8), and the mass ratio of the tannic acid to the polyvinyl pyrrolidone is (1-3):

1.

4. The method for preparing the bio-glue according to any one of claims 1 to 3, characterized in that: The steps include: The aqueous solution of polyvinyl pyrrolidone is mixed with the aqueous solution of gelatin to prepare solution A; and the aqueous solution of tannic acid is mixed with solution A to prepare bio-glue.

5. The preparation method according to claim 4, characterized in that: The polyvinyl pyrrolidone is mixed with water at a temperature of 60° C. to 80° C. and a speed of 500 rpm to 1000 rpm to prepare an aqueous solution of the polyvinyl pyrrolidone; Optionally, in the aqueous solution of polyvinyl pyrrolidone, the concentration of polyvinyl pyrrolidone is 10% w / v-30% w / v.

6. The preparation method according to claim 4 or 5, characterized in that: The gelatin is mixed with water at a temperature of 55°C to 65°C to prepare an aqueous solution of the gelatin; Optionally, the concentration of gelatin in the aqueous solution of gelatin is 2% w / v-10% w / v.

7. The preparation method according to claim 4 or 5, characterized in that: In the aqueous solution of tannic acid, the concentration of tannic acid is 10% w / v-30% w / v.

8. The preparation method according to claim 4 or 5, characterized in that: The conditions for mixing the aqueous solution of tannic acid with the solution A include stirring at a rate of 500 rpm to 1000 rpm for 18 h to 24 h.

9. The preparation method according to claim 4 or 5, characterized in that: After the tannic acid aqueous solution is mixed with the solution A, the mixture is centrifuged at 8000 rpm-10000 rpm for 5 min-10 min, and the obtained precipitate is washed with a PBS solution and then concentrated.

10. Use of the bioglue according to any one of claims 1 to 3 in the preparation of medical suture, injury repair or hemostatic materials.

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