Biological tissue adhesive as well as preparation method and application thereof

Abstract

The invention provides a preparation method of a biological tissue adhesive, which comprises the following steps: (1) respectively dissolving polyacrylic acid and polyethyleneimine in deionized water to prepare a polyacrylic acid aqueous solution and a polyethyleneimine aqueous solution; (2) blending the polyacrylic acid aqueous solution, the polyethyleneimine aqueous solution and the inorganic hemostatic material, and fully stirring to obtain a copolymer of the three substances; (3) immersing the copolymer into liquid nitrogen for pre-freezing, and then freeze-drying; and (4) taking out the freeze-dried sample, and grinding the sample into powder to obtain the biological tissue adhesive powder. The hemostatic adhesive powder is simple and convenient to apply, has a remarkable hemostatic effect and can be used for effectively stopping bleeding under various bleeding conditions.

Description

Technical Field

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

[0002] Uncontrollable massive bleeding is the main cause of death in wars, accidents and surgeries. Currently, surgical sutures, gauze packing, hemostatic sponges and other forms are commonly used in clinical practice to close wounds. However, these traditional hemostatic methods still have some defects. For example, traditional surgical sutures are complicated and time-consuming, cannot immediately close wounds, and are not suitable for hemostasis of incompressible soft tissues such as the heart and pancreas; gauze has poor moisture retention and is prone to adhesion to damaged tissues during use, and its hemostatic performance is often unstable when applied to irregular or deep wounds; hemostatic sponges usually require external force to stop bleeding when applied to complex wounds, which will cause secondary pain to patients; inorganic hemostatic powders can promote blood coagulation by absorbing water in the blood to concentrate red blood cells, platelets, and coagulation factors, but their tissue adhesion and integrity are poor during use, and they are easily washed away by the blood flow into the blood circulation, increasing the risk of vascular embolism and cytotoxicity.

[0003] In comparison, bioadhesives have been gradually used in clinical wound closure and hemostasis because of their simplicity, convenience, rapid hemostasis, and minimal tissue damage. Currently, a variety of bioadhesive hemostatic materials have been developed with the potential for hemostasis, including fibrin glue, cyanoacrylate glue, polyethylene glycol glue, etc. However, the cytotoxicity and weak tissue adhesion still limit the large-scale application of such hemostatic products. Summary of the invention

[0004] The purpose of the present invention is to provide a biological tissue adhesive and a preparation method and application thereof, aiming to solve the problem that the existing biological adhesive has cytotoxicity and weak tissue adhesion.

[0005] In order to solve the above technical problems, the present invention provides a method for preparing a biological tissue adhesive, comprising the following steps:

[0006] (1) dissolving polyacrylic acid and polyethyleneimine in deionized water respectively to prepare a polyacrylic acid aqueous solution with a mass fraction of 5 wt% to 15 wt% and a polyethyleneimine aqueous solution with a mass fraction of 10 wt% to 20 wt%;

[0007] (2) blending a polyacrylic acid aqueous solution, a polyethyleneimine aqueous solution and an inorganic hemostatic material in a mass ratio of (1-5):(1-10):20, and stirring thoroughly to obtain a copolymer of the three substances;

[0008] (3) immersing the copolymer in liquid nitrogen for pre-freezing (i.e., directly mixing with liquid nitrogen, the same below), and then freeze-drying;

[0009] (4) Taking out the freeze-dried sample and grinding it into powder to obtain the biological tissue adhesive.

[0010] Furthermore, the molecular weight of the polyacrylic acid is 3000-6000, and the molecular weight of the polyethyleneimine is 45000-70000.

[0011] Furthermore, the inorganic hemostatic material includes any one or more of zeolite, montmorillonite, kaolin, laponite, halloysite and mesoporous silica.

[0012] Furthermore, the particle size of the zeolite is less than 10um, the particle size of the montmorillonite is less than 1um, the particle size of the kaolin is less than 2um, the particle size of the laponite is less than 1.5um, the particle size of the halloysite is less than 2.5um, and the particle size of the mesoporous silica is less than 50nm.

[0013] Furthermore, the polyacrylic acid aqueous solution and the polyethyleneimine aqueous solution are prepared under the conditions of magnetic stirring at 500-800 rpm for 10-20 min at room temperature (25° C., the same below).

[0014] Furthermore, in step (4), the stirring time of the polyacrylic acid aqueous solution, the polyethyleneimine aqueous solution and the inorganic hemostatic material is 3-5 minutes, and the rotation speed is 300-400 rpm.

[0015] Furthermore, the pre-freezing time of the copolymer in liquid nitrogen is about 30-50 minutes, and the temperature is -196°C to -210°C.

[0016] Furthermore, the freeze drying of the copolymer is carried out in a vacuum freeze dryer, the drying time is 48-72 hours, the pressure is less than 5 Pa, and the temperature is lower than -55°C.

[0017] Furthermore, the particle size of the biological tissue adhesive is 160-220 meshes.

[0018] The invention also provides a biological tissue adhesive, which is prepared by the above-mentioned preparation method.

[0019] The present invention also provides application of the biological tissue adhesive in first aid hemostasis of wounds.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention prepares a biological tissue adhesive by encapsulating an inorganic hemostatic material in a biological tissue adhesive, which is mainly used for emergency hemostasis of wounds; specifically, when used, the hemostatic powder (i.e., biological tissue adhesive, also referred to as adhesive powder) can quickly absorb blood on the wound surface and form an adhesive physical barrier to block the bleeding wound, while the inorganic hemostatic material plays a procoagulant role, thereby achieving emergency hemostasis of the wound. At the same time, the hemostatic powder has strong tissue adhesion to different tissues and organs, and can effectively avoid the problems of vascular embolism and cytotoxicity caused by the use of a single inorganic hemostatic material, and has the characteristics of simple and easy operation, significant hemostatic effect, and good biological safety.

[0022] Moreover, the hemostatic powder provided by the present invention has stronger tissue adhesion than traditional commercially available fibrin glue and cyanoacrylate glue. The preparation method of the hemostatic powder material of the present application is simple and easy, the raw materials are easily available, the production cost is low, and it is suitable for industrial production. At the same time, the hemostatic powder material of the present application has a shorter hemostatic time, a more significant hemostatic effect, and better biological safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative labor.

[0024] Figure 1 This is a diagram showing the effect of a biological tissue adhesive prepared in Example 1 on hemostasis of rat tail amputation.

[0025] Figure 2 This is a diagram showing the effect of a biological tissue adhesive prepared in Example 2 on hemostasis of rat liver lacerations.

[0026] Figure 3 This is a Fourier transform infrared spectrum of a biological tissue adhesive prepared in Example 3.

[0027] Figure 4 Schematic diagram of tissue adhesion of a biological tissue adhesive prepared in Example 4.

[0028] Figure 5 This is a physical picture of a biological tissue adhesive prepared in Example 5. DETAILED DESCRIPTION

[0029] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments and drawings. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. The present invention is specifically introduced below in conjunction with the specific embodiments.

[0030] The embodiment of the present invention provides a method for preparing a biological tissue adhesive, comprising the following steps:

[0031] (1) dissolving polyacrylic acid and polyethyleneimine in deionized water respectively to prepare a polyacrylic acid aqueous solution with a mass fraction of 5 wt% to 15 wt% and a polyethyleneimine aqueous solution with a mass fraction of 10 wt% to 20 wt%;

[0032] (2) blending a polyacrylic acid aqueous solution, a polyethyleneimine aqueous solution and an inorganic hemostatic material in a mass ratio of (1-5):(1-10):20, and stirring thoroughly to obtain a copolymer of the three substances;

[0033] (3) pre-freezing the copolymer by immersing it in liquid nitrogen and then freeze-drying it;

[0034] (4) Taking out the freeze-dried sample and grinding it into powder to obtain the biological tissue adhesive.

[0035] Specifically, the molecular weight of the polyacrylic acid is 3000-6000, and the molecular weight of the polyethyleneimine is 45000-70000.

[0036] Specifically, the inorganic hemostatic material includes any one or more of zeolite, montmorillonite, kaolin, laponite, halloysite and mesoporous silica.

[0037] Specifically, the particle size of the zeolite is less than 10um, the particle size of the montmorillonite is less than 1um, the particle size of the kaolin is less than 2um, the particle size of the laponite is less than 1.5um, the particle size of the halloysite is less than 2.5um, and the particle size of the mesoporous silica is less than 50nm.

[0038] Specifically, the polyacrylic acid aqueous solution and the polyethyleneimine aqueous solution are prepared under the conditions of magnetic stirring at 500-800 rpm for 10-20 min at room temperature.

[0039] Specifically, in step (4), the stirring time of the polyacrylic acid aqueous solution, the polyethyleneimine aqueous solution and the inorganic hemostatic material is 3-5 minutes, and the rotation speed is 300-400 rpm.

[0040] Specifically, the pre-freezing time of the copolymer in liquid nitrogen is about 30-50 minutes, and the temperature is -196°C to -210°C.

[0041] Specifically, the freeze drying of the copolymer is carried out in a vacuum freeze dryer, the drying time is 48-72 hours, the pressure is less than 5 Pa, and the temperature is lower than -55°C.

[0042] Specifically, the particle size of the biological tissue adhesive is 160-220 meshes.

[0043] The embodiment of the present invention further provides a biological tissue adhesive, which is prepared by the above-mentioned preparation method.

[0044] The embodiment of the present invention also provides the use of the above biological tissue adhesive in first aid hemostasis of wounds.

[0045] Example 1

[0046] A method for preparing a biological tissue adhesive comprises the following steps: (1) at room temperature, adding polyacrylic acid with a molecular weight (MW) of 5000 and polyethyleneimine with a molecular weight (MW) of 70000 to deionized water respectively, stirring with a magnetic force at 500 rpm for 10 min, to prepare a polyacrylic acid aqueous solution with a mass fraction of 5wt% and a polyethyleneimine aqueous solution with a mass fraction of 10wt%.

[0047] (2) A polyacrylic acid aqueous solution with a mass fraction of 5 wt %, a polyethyleneimine aqueous solution with a mass fraction of 10 wt % and a zeolite with a particle size of 5 μm were blended in a mass ratio of 1:1:20, and the mixture was stirred at 300 rpm for 3 min to obtain a copolymer of the three substances.

[0048] (3) The copolymer was immersed in liquid nitrogen at -198°C for pre-freezing for 35 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 48 hours.

[0049] (4) Take out the freeze-dried sample and grind it into powder (mesh size is 160 mesh), and then obtain the biological tissue adhesive, that is, the biological tissue adhesive powder used for emergency hemostasis of wounds. The hemostasis effect is shown in the attached Figure 1 .

[0050] Example 2

[0051] A method for preparing a biological tissue adhesive comprises the following steps:

[0052] (1) At room temperature, polyacrylic acid with a molecular weight (MW) of 3000 and polyethyleneimine with a molecular weight (MW) of 50000 were added to deionized water, respectively, and magnetic stirring was performed at 600 rpm for 15 min to prepare a polyacrylic acid aqueous solution with a mass fraction of 10 wt% and a polyethyleneimine aqueous solution with a mass fraction of 15 wt%.

[0053] (2) A polyacrylic acid aqueous solution with a mass fraction of 10 wt %, a polyethyleneimine aqueous solution with a mass fraction of 15 wt % and montmorillonite with a particle size of 0.2 μm were blended in a mass ratio of 3:2:20, and the mixture was stirred at 350 rpm for 3 min to obtain a copolymer of the three substances.

[0054] (3) The copolymer was immersed in liquid nitrogen at -200°C for pre-freezing for 35 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 50 hours.

[0055] (4) Take out the freeze-dried sample and grind it into powder (mesh size is 180 mesh) to obtain biological tissue adhesive. The hemostatic effect is shown in the attached Figure 2 .

[0056] Example 3

[0057] A method for preparing a biological tissue adhesive comprises the following steps:

[0058] (1) At room temperature, polyacrylic acid with a molecular weight (MW) of 6000 and polyethyleneimine with a molecular weight (MW) of 55000 were added to deionized water, respectively, and magnetic stirring was performed at 550 rpm for 18 min to prepare a polyacrylic acid aqueous solution with a mass fraction of 8 wt % and a polyethyleneimine aqueous solution with a mass fraction of 10 wt %.

[0059] (2) A polyacrylic acid aqueous solution with a mass fraction of 8 wt %, a polyethyleneimine aqueous solution with a mass fraction of 10 wt % and kaolin with a particle size of 1 μm were blended in a mass ratio of 2:3:20, and the mixture was stirred at 400 rpm for 3 min to obtain a copolymer of the three substances.

[0060] (3) The copolymer was pre-frozen in liquid nitrogen at -210°C for 35 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 48 hours.

[0061] (4) Take out the freeze-dried sample and grind it into powder (mesh size is 200 mesh) to obtain a biological tissue adhesive. Its infrared spectrum is shown in the attached Figure 3 .

[0062] Example 4

[0063] A method for preparing a biological tissue adhesive comprises the following steps:

[0064] (1) At room temperature, polyacrylic acid with a molecular weight (MW) of 6000 and polyethyleneimine with a molecular weight (MW) of 70000 were added to deionized water, respectively, and magnetic stirring was performed at 550 rpm for 18 min to prepare a polyacrylic acid aqueous solution with a mass fraction of 5 wt% and a polyethyleneimine aqueous solution with a mass fraction of 20 wt%.

[0065] (2) A polyacrylic acid aqueous solution with a mass fraction of 5 wt %, a polyethyleneimine aqueous solution with a mass fraction of 20 wt % and hectorite with a particle size of 0.5 μm were mixed in a mass ratio of 2:3:20, and the mixture was stirred at 400 rpm for 5 min to obtain a copolymer of the three substances.

[0066] (3) The copolymer was immersed in liquid nitrogen at -200°C for pre-freezing for 40 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 50 hours.

[0067] (4) Take out the freeze-dried sample and grind it into powder (mesh size is 220 mesh) to obtain the biological tissue adhesive. The tissue adhesion effect diagram is shown in the attached figure. Figure 4 .

[0068] Example 5

[0069] A method for preparing a biological tissue adhesive comprises the following steps:

[0070] (1) At room temperature, polyacrylic acid with a molecular weight (MW) of 5000 and polyethyleneimine with a molecular weight (MW) of 50000 were added to deionized water, respectively, and magnetic stirring was performed at 550 rpm for 20 min to prepare a polyacrylic acid aqueous solution with a mass fraction of 10 wt% and a polyethyleneimine aqueous solution with a mass fraction of 10 wt%.

[0071] (2) A polyacrylic acid aqueous solution with a mass fraction of 10 wt %, a polyethyleneimine aqueous solution with a mass fraction of 10 wt % and halloysite with a particle size of 0.5 μm were blended in a mass ratio of 4:3:20, and the mixture was stirred at 300 rpm for 5 min to obtain a copolymer of the three substances.

[0072] (3) The copolymer was immersed in liquid nitrogen at -210°C for pre-freezing for 40 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 72 hours.

[0073] (4) Take out the freeze-dried sample and grind it into powder (mesh size is 200 mesh) to obtain the biological tissue adhesive. The actual picture is shown in the attached Figure 5 .

[0074] Example 6

[0075] A method for preparing a biological tissue adhesive comprises the following steps:

[0076] (1) At room temperature, polyacrylic acid with a molecular weight (MW) of 3000 and polyethyleneimine with a molecular weight (MW) of 70000 were added to deionized water, respectively, and magnetic stirring was performed at 500 rpm for 20 min to prepare a polyacrylic acid aqueous solution with a mass fraction of 15 wt% and a polyethyleneimine aqueous solution with a mass fraction of 15 wt%.

[0077] (2) A polyacrylic acid aqueous solution with a mass fraction of 10 wt %, a polyethyleneimine aqueous solution with a mass fraction of 10 wt % and mesoporous silica with a particle size of 20 nm were blended in a mass ratio of 4:1:20, and stirred at 400 rpm for 3 min to obtain a copolymer of the three substances.

[0078] (3) The copolymer was immersed in liquid nitrogen at -210°C for pre-freezing for 40 minutes, and then placed in a vacuum freeze dryer for freeze drying at a pressure of less than 5 Pa and a temperature below -55°C for 48 hours.

[0079] (4) Taking out the freeze-dried sample and grinding it into powder (mesh size is 160 mesh) to obtain the biological tissue adhesive.

[0080] The above description is only a description of the preferred embodiments of the present invention, and is not intended to limit the scope of the present invention. Any changes or modifications made by a person skilled in the art in the field of the present invention based on the above disclosure shall fall within the scope of protection of the claims.

Claims

1. A method for preparing a biological tissue adhesive, characterized in that: The following steps are involved: (1) dissolving polyacrylic acid and polyethyleneimine in deionized water respectively to prepare a polyacrylic acid aqueous solution with a mass fraction of 5 wt% to 15 wt% and a polyethyleneimine aqueous solution with a mass fraction of 10 wt% to 20 wt%; (2) blending a polyacrylic acid aqueous solution, a polyethyleneimine aqueous solution and an inorganic hemostatic material in a mass ratio of (1-5):(1-10):20, and stirring thoroughly to obtain a copolymer of the three substances; (3) pre-freezing the copolymer by immersing it in liquid nitrogen and then freeze-drying it; (4) Taking out the freeze-dried sample and grinding it into powder to obtain the biological tissue adhesive.

2. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: The molecular weight of the polyacrylic acid is 3000-6000, and the molecular weight of the polyethyleneimine is 45000-70000.

3. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: The inorganic hemostatic material includes any one or more of zeolite, montmorillonite, kaolin, laponite, halloysite and mesoporous silica.

4. The method for preparing a biological tissue adhesive according to claim 3, characterized in that: The particle size of the zeolite is less than 10um, the particle size of the montmorillonite is less than 1um, the particle size of the kaolin is less than 2um, the particle size of the laponite is less than 1.5um, the particle size of the halloysite is less than 2.5um, and the particle size of the mesoporous silica is less than 50nm.

5. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: The polyacrylic acid aqueous solution and the polyethyleneimine aqueous solution are prepared under the conditions of magnetic stirring at 500-800 rpm for 10-20 min at room temperature.

6. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: In step (4), the stirring time of the polyacrylic acid aqueous solution, the polyethyleneimine aqueous solution and the inorganic hemostatic material is 3-5 minutes, and the rotation speed is 300-400 rpm.

7. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: The pre-freezing time of the copolymer in liquid nitrogen is about 30-50 minutes, and the temperature is -196°C to -210°C; And / or, the freeze drying of the copolymer is carried out in a vacuum freeze dryer, the drying time is 48-72 hours, the pressure is less than 5 Pa, and the temperature is lower than -55°C.

8. The method for preparing a biological tissue adhesive according to claim 1, characterized in that: The particle size of the biological tissue adhesive is 160-220 meshes.

9. A biological tissue adhesive, characterized in that: The biological tissue adhesive is prepared by the preparation method of any one of claims 1 to 8.

10. Use of the biological tissue adhesive according to claim 9 in first aid hemostasis of wounds.

Images