Hydrogel for bonding biological tissues as well as preparation method and application of hydrogel

Abstract

The invention relates to the technical field of medical materials, in particular to hydrogel for biological tissue bonding and a preparation method and application thereof. The hydrogel for biological tissue adhesion comprises a first component, a second component, a third component and a fourth component, the first component and the second component form electrostatic interaction, and the third component and the fourth component form a covalent cross-linked structure; the first component, the second component, the third component and the fourth component are all biocompatible components. The hydrogel for biological tissue adhesion provided by the invention has excellent biocompatibility, relatively high adhesion strength and relatively low swelling property.

Description

Technical Field

[0001] The present application relates to the technical field of medical materials, and in particular to a biological tissue adhesive hydrogel and a preparation method and application thereof. Background Art

[0002] Vascular anastomosis is a very common operation in vascular surgery and thoracic surgery. The most mainstream and basic technology is the interrupted suture anastomosis, which is to connect the two ends of the blood vessels with surgical sutures. It is regarded as the "gold standard" of surgery. However, this method has the following problems: (1) The operation is cumbersome, resulting in a long operation time. If it involves open-chest surgery, the patient will stay in the open-chest state for too long, thereby increasing the risk of surgery; (2) To achieve complete no bleeding at the anastomosis site, especially for large blood vessels such as the aorta, more sutures and smaller suture needle insertion intervals are required, and the operation is more rigorous; (3) In addition to bleeding caused by loose sutures at the anastomosis, the size of the hole left by the suture needle penetrating the blood vessel wall is usually larger than the size of the suture. The resulting gap will also cause bleeding under the action of blood pressure, especially when the surgical site is on large blood vessels such as the aorta.

[0003] To solve the above problems, researchers and clinicians have proposed many methods, such as mechanical anastomosis, sleeve anastomosis and vascular bonding anastomosis. Among them, vascular bonding anastomosis is to use biological tissue adhesives with adhesion ability combined with sutures or simply rely on adhesives to complete vascular anastomosis. Currently common biological tissue adhesives include synthetic glue, biological glue and semi-synthetic glue. Common types of synthetic glues include cyanoacrylates and polyethylene glycol hydrogels, common types of biological glues are fibrin glues, and semi-synthetic glues are usually albumin or gelatin adhesives. However, the biological tissue adhesives currently used have the following problems: the main components of biological glue, such as fibrin and albumin, are mainly extracted from animals such as pigs and cattle. Such animal-derived products have potential risks of disease transmission, such as mad cow disease, and the strength of biological glue currently on the market is weaker than other types of tissue adhesives; semi-synthetic glue is usually based on the mutual reaction of protein and polyaldehyde compounds (such as albumin and glutaraldehyde) to form glue. This type of sealant also has the risk of disease transmission brought by animal-derived materials. At the same time, the residual polyaldehyde compounds will have a toxic effect on surrounding tissues and even the whole body; synthetic glue, such as cyanoacrylates, will release a lot of heat locally during the polymerization process, which has potential damage to surrounding tissues. Moreover, after the bonding is completed, the polymer formed is relatively hard and has poor compliance with soft tissues such as blood vessels. At the same time, the small molecules produced during the degradation process and the residual monomers wrapped inside are biologically toxic. Polyethylene glycol-based hydrogel sealants will swell in the body fluid environment, and there is a risk of compressing blood vessels and surrounding tissues and organs. Summary of the invention

[0004] Based on this, it is necessary to provide a biological tissue adhesive hydrogel and a preparation method and application thereof. The hydrogel has excellent biocompatibility, high adhesion strength and low swelling property.

[0005] In a first aspect, the present application provides a hydrogel for biological tissue adhesion, comprising: a first component, a second component, a third component and a fourth component; the first component and the second component form an electrostatic interaction, and the third component and the fourth component form a covalent cross-linked structure; the first component, the second component, the third component and the fourth component are all biocompatible components.

[0006] In some embodiments, the first component includes a positively charged group, and the positively charged group includes one or more of a primary amine group, a secondary amine group, a tertiary amine group, a quaternary ammonium group, an imidazole group, a guanidine group, and a pyridine group;

[0007] The second component includes negatively charged groups, and the negatively charged groups include one or more of hydroxyl groups, carboxyl groups, sulfonic acid groups and phosphate groups;

[0008] The electrostatic interaction is formed between the positively charged group and the negatively charged group.

[0009] In some embodiments, the first component includes one or more of polyethyleneimine, polyallylamine, polyethylene glycol modified with the positively charged group, polysaccharide containing or modified with the positively charged group, and polyamino acid containing or modified with the positively charged group;

[0010] The second component includes one or more of polyacrylic acid and its water-soluble derivatives, polyethylene glycol modified with the negatively charged group, polysaccharide containing or modified with the negatively charged group, and polyamino acid containing or modified with the negatively charged group.

[0011] In some embodiments, the third component includes a first reactive group, and the first reactive group includes one or more of a hydroxyl group, a thiol group, an amino group, an imino group, and an azide group;

[0012] The fourth component includes a second reactive group, and the second reactive group includes one or more of anhydride, aldehyde, succinimide, maleimide, carbon-carbon double bond, carbon-carbon triple bond, isocyanate, epoxy, carboxyl and isothiocyanate;

[0013] The first reactive group and the second reactive group can be chemically cross-linked.

[0014] In some embodiments, the third component includes one or more of polyethyleneimine, polyamino acid, water-soluble protein, gelatin, and polyethylene glycol modified with the first active group;

[0015] The fourth component includes one or both of polyethylene glycol modified with the second active group and polysaccharide modified with the second active group.

[0016] In some embodiments, the number average molecular weight of the first component and the second component are independently 1 kDa to 2×10 3 kDa, the number average molecular weight of the third component and the fourth component are independently 100Da to 2×10 3 kDa.

[0017] In some embodiments, the ratio of the sum of the mass of the first component and the second component to the sum of the mass of the third component and the fourth component is 1:(1-1000);

[0018] and / or, the mass ratio of the first component to the second component is 1:(0.05-20);

[0019] And / or, the mass ratio of the third component to the fourth component is 1:(0.02-50).

[0020] In a second aspect, the present application provides a method for preparing the hydrogel as described in the first aspect, comprising the following steps:

[0021] dissolving the first component and the third component in a first solvent to form a first solution;

[0022] dissolving the second component and the fourth component in a second solvent to form a second solution;

[0023] The first solution and the second solution are mixed so that the first component and the second component form a non-covalent cross-linked network, and the third component and the fourth component form a covalent cross-linked network, thereby preparing the hydrogel.

[0024] In some embodiments, the preparation method satisfies at least one of the following characteristics:

[0025] 1) In the first solution, the concentration of the first component is 0.05 w / v% to 1 w / v%, and the concentration of the third component is 1 w / v% to 50 w / v%;

[0026] 2) In the second solution, the concentration of the second component is 0.05 w / v% to 1 w / v%, and the concentration of the fourth component is 1 w / v% to 50 w / v%;

[0027] 3) The volume ratio of the first solution to the second solution is (0.01-1):(1-0.01).

[0028] In some embodiments, the pH of the first solvent is 1≤pH<7, and the pH of the second solvent is 7≤pH≤13.

[0029] In some embodiments, the first solvent comprises one or more of hydrochloric acid solution, sulfuric acid solution, phosphoric acid solution, phosphoric acid-phosphate buffer, citric acid-citrate buffer, boric acid-borate buffer, acetic acid-acetate buffer, glycine-hydrochloric acid buffer, amino acid buffer and amino acid salt buffer;

[0030] And / or, the second solvent includes one or more of pure water, physiological saline, sodium hydroxide solution, potassium hydroxide solution, phosphate buffer, carbonate buffer, borate buffer, acetate buffer, citrate buffer, glycine-sodium hydroxide buffer, amino acid buffer and amino acid salt buffer solution.

[0031] In a third aspect, the present application provides use of the hydrogel described in the first aspect in preparing a medical sealant or as an in vitro medical sealant.

[0032] The biological tissue adhesive hydrogel provided in the present application has a multiple cross-linked network structure formed by electrostatic interactions and covalent cross-linked structures. Compared with a single non-covalent cross-linked structure, the hydrogel has better operability when formed, and the prepared hydrogel has higher mechanical properties, adhesion strength and controllability; the formation speed of electrostatic interactions is often faster than the formation of covalent cross-linked structures. Therefore, compared with a single covalent cross-linked structure, the hydrogel with a multiple cross-linked network structure provided in the present application can quickly reach a higher viscosity when two solutions are mixed, so that it can stably adhere to the applied position without flowing arbitrarily to wait for the covalent cross-linked network to form to achieve higher strength.

[0033] At the same time, the hydrogel with a multi-crosslinked network structure has higher toughness, and the presence of the multi-crosslinked network structure can also make the hydrogel swelling rate lower and prolong its degradation time. In addition, the hydrogel provided by the present application has good antibacterial properties due to electrostatic interaction and contains positively charged groups, and can effectively prevent infection after being applied to the corresponding parts. Therefore, the hydrogel provided by the present application is very conducive to the preparation of medical sealants for vascular occlusion or tissue adhesion and hemostasis.

[0034] In addition, in the hydrogel provided by the present application, the electrostatic interaction and the covalent cross-linking structure are formed by different components respectively. Compared with the hydrogel based on two components to realize the electrostatic interaction and the covalent cross-linking structure, the composition of the electrostatic network and the covalent network can be regulated as needed, so the formation of these two structures can be independently regulated, and the hydrogel with the target performance can be prepared more controllably and conveniently. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0036] Figure 1 This is a process flow chart for preparing a hydrogel in one embodiment of the present application;

[0037] Figure 2 Schematic diagram of the structure of the device used to test the adhesion strength of hydrogels. DETAILED DESCRIPTION

[0038] In order to facilitate the understanding of the present application, the present application will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present application are given in the drawings. However, the present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive.

[0039] 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.

[0040] the term

[0041] The term "and / or" used herein includes any and all combinations of one or more of the related listed items. For example, "A and / or B" includes three parallel solutions of A, B and "a combination of A and B".

[0042] The term "one or more" used herein means any one of the listed items or any combination of the listed items unless otherwise specified. Similarly, "one or more" and the like in other cases where they mean "one or more" are also understood in the same way unless otherwise specified.

[0043] The terms "further", "further", "particularly", "for example", "such as", "example", "for example" and the like used herein are used for descriptive purposes, indicating that the previous and subsequent different technical solutions are related in terms of the contents covered, but should not be understood as limiting the previous technical solution, nor can they be understood as limiting the scope of protection of the present invention. In the present invention, unless otherwise specified, A (such as B) means that B is a non-limiting example of A, and it can be understood that A is not limited to B.

[0044] In the terms "first aspect", "second aspect", "third aspect", "fourth aspect", etc. used herein, the terms "first", "second", "third", "fourth", etc. are used only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0045] As used herein, the terms "comprising," "including," and "comprising" are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or method steps.

[0046] In this article, if there are multiple steps involved in the method flow, unless there is a clear different description in this article, there is no strict order restriction for the execution of these steps, and they can be executed in other orders than described. Moreover, any step can include multiple sub-steps or multiple stages, and these sub-steps or stages are not necessarily executed at the same time, but can be executed at different times, and their execution order is not necessarily sequential, but can be executed in turn, alternating or simultaneously with other steps or parts of sub-steps or stages of other steps.

[0047] Traditional sealants used for blood vessel occlusion or hemostasis have certain defects. For example, biological glue has a potential risk of disease transmission; semi-synthetic glues based on proteins and polyaldehyde compounds not only have the risk of disease transmission, but also have toxic effects on organisms; and cyanoacrylate synthetic glues not only damage organisms, but also have high hardness and poor compliance with soft tissues such as blood vessels. In addition, the currently used polyethylene glycol-based hydrogel sealants are prone to swelling, which will compress surrounding tissues, organs and blood vessels. To this end, the present application provides a biological tissue bonding hydrogel, which has excellent biocompatibility, high adhesion strength and low swelling rate, so that it can be used to prepare medical sealants for blood vessel occlusion or hemostasis.

[0048] In a first aspect, the present application provides a biological tissue adhesion hydrogel, which includes: a first component, a second component, a third component and a fourth component; the first component and the second component form an electrostatic interaction, and the third component and the fourth component form a covalent cross-linked structure; the first component, the second component, the third component and the fourth component are all biocompatible components.

[0049] The biological tissue adhesive hydrogel provided in the present application has a multiple cross-linked network structure formed by electrostatic interactions and covalent cross-linked structures. Compared with a single non-covalent cross-linked structure, the hydrogel has better operability when formed, and the prepared hydrogel has higher mechanical properties, adhesion strength and controllability; the formation speed of electrostatic interactions is often faster than the formation of covalent cross-linked structures. Therefore, compared with a single covalent cross-linked structure, the hydrogel with a multiple cross-linked network structure provided in the present application has a higher viscosity, so that it can stably adhere to the applied position without flowing at will.

[0050] At the same time, the hydrogel with a multi-crosslinked network structure has higher toughness, and the presence of the multi-crosslinked network structure can also make the hydrogel swelling rate lower and prolong its degradation time. In addition, the hydrogel provided by the present application has good antibacterial properties due to electrostatic interaction and contains positively charged groups, and can effectively prevent infection after being applied to the corresponding parts. Therefore, the hydrogel provided by the present application is very conducive to the preparation of medical sealants for vascular occlusion or tissue adhesion and hemostasis.

[0051] In addition, in the hydrogel provided by the present application, the electrostatic interaction and the covalent cross-linking structure are formed by different components, respectively, so the formation of these two structures can be independently regulated, and the hydrogel with target properties can be prepared in a more controllable and convenient manner.

[0052] It can be understood that the biocompatible component specifically refers to the compatibility between the material and the host, the material has the characteristics of being non-biotoxic and can cause appropriate reactions in specific parts of the body.

[0053] In some embodiments, the biocompatible component includes one or more of polysaccharides and their derivatives, water-soluble proteins and their derivatives, polyacrylic acid and their water-soluble derivatives, polyethylene glycol and its derivatives, water-soluble amino polymers and polyamino acids.

[0054] In some embodiments, the water-soluble amino polymer includes one or more of polyethyleneimine and polyallylamine.

[0055] In some embodiments, the polysaccharide includes one or more of chitosan, hyaluronic acid, dextran, alginate, cellulose, starch, chondroitin sulfate, heparin, carrageenan, agarose, xanthan gum, and pullulan.

[0056] In some embodiments, the water-soluble protein includes one or more of fibrin, collagen, albumin, silk fibroin, and elastin.

[0057] In some embodiments, the first component includes a positively charged group, wherein the first component includes a positively charged group, and the positively charged group includes one or more of a primary amine group, a secondary amine group, a tertiary amine group, a quaternary ammonium group, an imidazole group, a guanidine group, and a pyridine group;

[0058] The second component includes negatively charged groups, which include one or more of hydroxyl, carboxyl, sulfonic acid and phosphate groups;

[0059] Electrostatic interactions are formed between the positively charged groups and the negatively charged groups.

[0060] It can be understood that the first component and the second component with positively charged groups can be ionized in water and form positively charged and negatively charged structures respectively, thereby forming cross-linking effects inside the hydrogel through electrostatic effects.

[0061] In some embodiments, the first component is selected from a group consisting of 3 Preferably, the first component is selected from polymers with a number average molecular weight of 3 kDa to 1500 kDa.

[0062] In some embodiments, the first component includes one or more of polyethyleneimine, polyallylamine, polyethylene glycol modified with positively charged groups, polysaccharides containing or modified with positively charged groups, and polyamino acids containing or modified with positively charged groups.

[0063] In some embodiments, the second component is selected from a group consisting of 3 Preferably, the second component is selected from polymers with a number average molecular weight of 10 kDa to 800 kDa.

[0064] In some embodiments, the second component includes one or more of polyacrylic acid and its water-soluble derivatives, polyethylene glycol modified with negatively charged groups, polysaccharides containing or modified with negatively charged groups, and polyamino acids containing or modified with negatively charged groups.

[0065] In some embodiments, the third component includes a first reactive group, and the first reactive group includes one or more of a hydroxyl group, a thiol group, an amino group, an imino group, and an azide group;

[0066] The fourth component includes a second reactive group, and the second reactive group includes one or more of anhydride, aldehyde, succinimide, maleimide, carbon-carbon double bond, carbon-carbon triple bond, isocyanate, epoxy, carboxyl and isothiocyanate;

[0067] The first reactive group and the second reactive group can be chemically cross-linked.

[0068] In some embodiments, the third component and the fourth component are independently selected from a number average molecular weight of 100Da to 2×10 3 Preferably, the third component and the fourth component are independently selected from polymers with a number average molecular weight of 100Da to 5×10 2 kDa small molecule compounds or polymers.

[0069] In some embodiments, the third component includes one or more of polyethyleneimine, polyamino acid, water-soluble protein, gelatin and polyethylene glycol modified with a first active group. Among them, small molecule compounds include polyethyleneimine and / or polyamino acid with a molecular weight lower than 1000Da, such as polylysine with a degree of polymerization of 3 to 5; polymers include polyethyleneimine, polylysine, polyethylene glycol modified with a first active group, such as polyethylene glycol modified with an amino group, polyethylene glycol modified with a thiol group, with a molecular weight higher than 1000Da.

[0070] In some embodiments, the fourth component includes one or both of polyethylene glycol modified with a second active group and a polysaccharide modified with a second active group. Among them, the small molecule compound includes polyethylene glycol modified with a second active group (molecular weight less than 1000Da), such as polyethylene glycol acrylate, polyethylene glycol succinic acid succinimidyl ester; the polymer includes one or more of polyethylene glycol modified with a second active group (molecular weight greater than 1000Da) and a polysaccharide.

[0071] Compared with polymers (high molecular weight compounds), small molecular compounds have higher reactivity and the cross-linking density of the formed hydrogel is greater. Therefore, when the third component and the fourth component are both selected from small molecular compounds, the formed hydrogel has a higher compression modulus and adhesion strength, a faster gelation speed, and a lower swelling ratio.

[0072] It can be understood that the polyethylene glycol involved in the first component, the second component, the third component and the fourth component can specifically be four-arm polyethylene glycol, six-arm polyethylene glycol or eight-arm polyethylene glycol.

[0073] In some embodiments, the ratio of the sum of the mass of the first component and the second component to the sum of the mass of the third component and the fourth component is 1: (1-1000), for example, 1:900, 1:800, 1:500, 1:200, 1:100, 1:80, 1:75, 1:70, 1:60, 1:50, 1:30, 1:25, 1:20, 1:10, 1:5. By regulating the ratio of the sum of the mass of the first component and the second component to the sum of the mass of the third component and the fourth component, the ratio of the non-covalent cross-linked structure and the covalent cross-linked structure can be regulated, thereby more flexibly regulating the properties of the hydrogel without the need for complex chemical synthesis and precisely designed physical processes. Moreover, by regulating the ratio within the above range, the construction of hydrogel can be better realized; if the ratio is higher than this range, it is easy to cause the components carrying positive and negative charges to combine too quickly, resulting in static electricity not being evenly distributed inside the hydrogel, and even phase separation causing precipitation; and if the ratio is lower than this range, it is easy to cause the non-covalent electrostatic cross-linking network to have too little effect, making it difficult to affect the overall performance of the hydrogel.

[0074] In some embodiments, the mass ratio of the first component to the second component is 1:(0.05-50), for example, 1:40, 1:30, 1:20, 1:10, 1:5, 1:2, 1:1, 1:0.5, 1:0.2, 1:0.1.

[0075] In some embodiments, the mass ratio of the third component to the fourth component is 1:(0.05-50), for example, 1:40, 1:30, 1:20, 1:10, 1:5, 1:2, 1:1, 1:0.5, 1:0.2, 1:0.1.

[0076] By regulating the mass ratio of the first component and the second component within the above range, the components carrying positive and negative charges can effectively achieve electrostatic crosslinking without sedimentation; regulating the mass ratio of the third component to the fourth component within the above range can allow the active groups to fully react to form a covalent crosslinking network.

[0077] See also Figure 1 In a second aspect, the present application provides a method for preparing the hydrogel as described in the first aspect, comprising the following steps:

[0078] S100: dissolving the first component and the third component in a first solvent to form a first solution;

[0079] S200: dissolving the second component and the fourth component in a second solvent to form a second solution;

[0080] S300: mixing the first solution and the second solution so that the first component and the second component form a non-covalent cross-linked network, and the third component and the fourth component form a covalent cross-linked network, to prepare a hydrogel.

[0081] Step S100: forming a first solution.

[0082] In some embodiments, in the first solution, the concentration of the first component is 0.05w / v% to 1w / v%, for example, 0.1w / v%, 0.2w / v%, 0.25w / v%, 0.3w / v%, 0.5w / v%, 0.8w / v%; the concentration of the third component is 1w / v% to 50w / v%, for example, 2w / v%, 5w / v%, 10w / v%, 15w / v%, 20w / v%, 30w / v%, 40w / v%.

[0083] In some embodiments, the pH of the first solvent is 1≤pH<7, for example, the pH of the first solvent is 1, 2, 3, 4, 5, or 6. By adjusting the pH of the first solvent within the above range, the positive charge of the first component can be ensured.

[0084] In some embodiments, the first solvent includes one or more of an inorganic acid solution, phosphoric acid-phosphate buffer, citric acid-citrate buffer, boric acid-borate buffer, acetic acid-acetate buffer, glycine-hydrochloric acid buffer, amino acid buffer and amino acid salt buffer. Wherein, the inorganic acid solution includes one or more of a hydrochloric acid solution, a sulfuric acid solution and a phosphoric acid solution. Further, the pH value of the inorganic acid solution is 1 to 6.

[0085] Step S200: forming a second solution.

[0086] In some embodiments, in the second solution, the concentration of the second component is 0.05w / v% to 1w / v%, for example, 0.1w / v%, 0.15w / v%, 0.2w / v%, 0.5w / v%, 0.8w / v%; the concentration of the fourth component is 1w / v% to 50w / v%, for example, 5w / v%, 10w / v%, 15w / v%, 20w / v%, 30w / v%, 40w / v%.

[0087] In some embodiments, the second solvent has a pH of 7≤pH≤13, for example, the first solvent has a pH of 7, 8, 9, 10, 11, 12, or 13. By adjusting the pH of the second solvent within the above range, the negative charge of the second component can be ensured.

[0088] In some embodiments, the second solvent includes pure water, physiological saline, inorganic alkaline solution, phosphate buffer, carbonate buffer, borate buffer, acetate buffer, citrate buffer, glycine-sodium hydroxide buffer, amino acid buffer or amino acid salt buffer solution. Among them, the inorganic alkaline solution includes one or more of sodium hydroxide solution and potassium hydroxide solution; further, the pH of the inorganic alkaline solution is 8-12.

[0089] Step S300: preparing a hydrogel.

[0090] In some embodiments, the volume ratio of the first solution to the second solution is (0.01-1):(1-0.01), for example, 1:0.03, 1:0.5, 1:1, 0.5:1, 0.05:1.

[0091] In some embodiments, after the first solution and the second solution are mixed, a step of standing is further included. The standing time and temperature are not limited, and are based on the condition that the first component and the second component can form a non-covalent cross-linked network, and the third component and the fourth component can form a covalent cross-linked network. For example, the standing time can be 1 second to 8 hours, and the standing temperature can be room temperature.

[0092] It should be noted that, in the present application, "room temperature" generally refers to 4°C to 35°C, for example, 20°C ± 5°C. In some embodiments of the present application, "room temperature" refers to 10°C to 30°C. In some embodiments of the present application, "room temperature" refers to 20°C to 30°C.

[0093] In a third aspect, the present application provides use of the hydrogel described in the first aspect in preparing a medical sealant or as an in vitro medical sealant.

[0094] In the present application, medical sealants are mainly used to prevent bleeding in various parts of the body, bleeding after blood vessel suture, or for blood vessel occlusion.

[0095] The present application is further described in detail below in conjunction with specific embodiments.

[0096] Example 1

[0097] 1) dissolving 4 g of aldehyde-terminated four-arm polyethylene glycol with a number average molecular weight of 20 kDa and 0.1 g of sodium alginate with a number average molecular weight of 100 kDa in 20 mL of phosphate buffer with a pH of 6 to form a first solution;

[0098] 2) dissolving 1 g of hydroxypropyl chitosan with a number average molecular weight of 100 kDa and 0.5 g of polyarginine with a number average molecular weight of 2 kDa in 20 mL of sodium carbonate-sodium bicarbonate buffer with a pH of 9 to form a second solution;

[0099] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:0.03 and mixed, and allowed to stand for 5 minutes to obtain a hydrogel. In this embodiment, the hydrogel has a multiple cross-linked network structure composed of a covalent cross-linked structure formed by four-arm polyethylene glycol and hydroxypropyl chitosan, a covalent cross-linked structure formed by four-arm polyethylene glycol and polyarginine, and a non-covalent cross-linked structure (electrostatic interaction) formed by hydroxypropyl chitosan (providing amino groups) and sodium alginate (providing carboxyl groups).

[0100] Example 2

[0101] 1) dissolving 2 g of four-arm polyethylene glycol succinimidyl glutarate having a number average molecular weight of 20 kDa and 0.1 g of polyacrylic acid having a weight average molecular weight of 10 kDa in 20 mL of dilute hydrochloric acid having a pH of 2 (the molar concentration is 0.01 mol / L) to form a first solution;

[0102] 2) dissolving 2 g of polyethyleneimine with a number average molecular weight of 8 kDa and 0.1 g of quaternized chitosan with a number average molecular weight of 80 kDa in 20 mL of a sodium hydroxide solution with a pH of 9 to form a second solution;

[0103] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:0.5, mixed, and allowed to stand for 10 minutes to obtain a hydrogel. In this embodiment, the hydrogel has a multi-crosslinked network structure composed of a non-covalent crosslinked structure (electrostatic interaction) formed by polyacrylic acid and quaternized chitosan, a non-covalent crosslinked structure (electrostatic interaction) formed by polyacrylic acid and polyethyleneimine, and a covalent crosslinked structure formed by four-arm polyethylene glycol succinimidyl glutarate and polyethyleneimine.

[0104] Example 3

[0105] 1) dissolving 6 g of four-arm polyethylene glycol maleimide with a number average molecular weight of 10 kDa and 0.02 g of carboxymethyl cellulose with a number average molecular weight of 80 kDa in 20 mL of dilute hydrochloric acid with a pH of 3 (the molar concentration is 0.001 mol / L) to form a first solution;

[0106] 2) dissolving 6 g of thiol-terminated four-arm polyethylene glycol with a number average molecular weight of 20 kDa and 0.02 g of polylysine with a number average molecular weight of 3 kDa in 20 mL of phosphate-sodium hydroxide buffer with a pH of 9 to form a second solution;

[0107] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:1 and mixed, and allowed to stand for 20 minutes to obtain a hydrogel. In this embodiment, the hydrogel has a multi-crosslinked network structure composed of a non-covalent crosslinked structure (electrostatic interaction) formed by carboxymethyl cellulose and polylysine, a covalent crosslinked structure formed by four-arm polyethylene glycol maleimide and thiol-terminated four-arm polyethylene glycol, and a covalent crosslinked structure formed by four-arm polyethylene glycol maleimide and polylysine.

[0108] Example 4

[0109] 1) dissolving 3 g of four-arm polyethylene glycol methacrylate with a number average molecular weight of 10 kDa and 0.04 g of carboxymethyl dextran with a number average molecular weight of 80 kDa in 20 mL of dilute hydrochloric acid with a pH of 6 to form a first solution;

[0110] 2) dissolving 3 g of amino-terminated four-arm polyethylene glycol with a number average molecular weight of 10 kDa and 0.05 g of polylysine with a number average molecular weight of 3 kDa in 20 mL of phosphate buffer with a pH of 9 to form a second solution;

[0111] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 0.5:1 and mixed, and allowed to stand for 40 minutes to obtain a hydrogel. In this embodiment, the hydrogel contains a multi-crosslinked network structure composed of a non-covalent crosslinked structure (electrostatic interaction) formed by carboxymethyl dextran and polylysine, a non-covalent crosslinked structure (electrostatic interaction) formed by carboxymethyl dextran and amino-terminated four-arm polyethylene glycol, a covalent crosslinked structure formed by four-arm polyethylene glycol methacrylate and amino-terminated four-arm polyethylene glycol, and a covalent crosslinked structure formed by four-arm polyethylene glycol methacrylate and polylysine.

[0112] Example 5

[0113] 1) dissolving 5 g of four-arm polyethylene glycol succinimidyl glutarate having a number average molecular weight of 10 kDa and 0.05 g of carboxymethyl cellulose having a number average molecular weight of 100 kDa in 20 mL of dilute hydrochloric acid having a pH of 3 (the molar concentration is 0.001 mol / L) to form a first solution;

[0114] 2) dissolving 5 g of amino-terminated four-arm polyethylene glycol with a number average molecular weight of 10 kDa and 0.05 g of polylysine with a number average molecular weight of 2 kDa in 20 mL of phosphate-sodium hydroxide buffer with a pH of 9 to form a second solution;

[0115] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:1 and mixed, and allowed to stand for 0.5 min to obtain a hydrogel. In this embodiment, the hydrogel contains a multi-crosslinked network structure composed of a non-covalent crosslinked structure (electrostatic interaction) formed by carboxymethyl cellulose and polylysine, a non-covalent crosslinked structure (electrostatic interaction) formed by carboxymethyl cellulose and amino-terminated four-arm polyethylene glycol, a covalent crosslinked structure formed by four-arm polyethylene glycol succinimidyl glutarate and amino-terminated four-arm polyethylene glycol, and a covalent crosslinked structure formed by four-arm polyethylene glycol succinimidyl glutarate and polylysine.

[0116] Example 6

[0117] 1) 0.01 g of a number average molecular weight of 10 3 kDa sodium hyaluronate and 10 g glutaraldehyde are dissolved in 10 mL of phosphate-phosphate buffer at pH 6 to form a first solution;

[0118] 2) dissolving 1 g of polyethyleneimine with a number average molecular weight of 25 kDa and 0.05 g of quaternized dextran with a number average molecular weight of 50 kDa in 10 mL of phosphate buffer with a pH of 8 to form a second solution;

[0119] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 0.05:1 and mixed, and allowed to stand for 1 hour to obtain a hydrogel. In this embodiment, the hydrogel has a multi-crosslinked network structure composed of a non-covalent cross-linked structure (electrostatic interaction) formed by sodium hyaluronate and polyethyleneimine, a non-covalent cross-linked structure (electrostatic interaction) formed by sodium hyaluronate and quaternized dextran, and a covalent cross-linked structure formed by glutaraldehyde and polyethyleneimine.

[0120] Example 7

[0121] The preparation method of this embodiment is basically the same as that of embodiment 5, except that the masses of carboxymethyl cellulose and polylysine are 0.1 g and 0.2 g, respectively.

[0122] Comparative Example 1

[0123] The preparation method of this comparative example is basically the same as that of Example 2, except that the hydrogel prepared in this comparative example only has a covalent cross-linked structure. The specific steps are as follows:

[0124] 1) dissolving 2 g of four-arm polyethylene glycol succinimidyl glutarate having a number average molecular weight of 20 kDa in 20 mL of dilute hydrochloric acid having a pH of 2 (the molar concentration is 0.01%) to form a first solution;

[0125] 2) dissolving 2 g of polyethyleneimine having a number average molecular weight of 10 kDa in 20 mL of a sodium hydroxide solution having a pH of 9 to form a second solution;

[0126] 3) The first solution prepared in step 1) and the second solution prepared in step 2) were placed in a plastic culture dish at a volume ratio of 1:0.5, mixed, and allowed to stand for 10 minutes to prepare a hydrogel. In this embodiment, a covalent cross-linked structure formed by four-arm polyethylene glycol succinimidyl glutarate and polyethyleneimine exists inside the hydrogel.

[0127] Comparative Example 2

[0128] The preparation method of this comparative example is basically the same as that of Example 3, except that the hydrogel prepared in this comparative example only has a covalent cross-linked structure. The specific steps are as follows:

[0129] 1) dissolving 6 g of four-arm polyethylene glycol maleimide having a number average molecular weight of 10 kDa in 20 mL of dilute hydrochloric acid having a pH of 3 (amount of substance concentration of 0.001) to form a first solution;

[0130] 2) dissolving 6 g of thiol-terminated four-arm polyethylene glycol with a number average molecular weight of 20 kDa in 20 mL of phosphate-sodium hydroxide buffer with a pH of 9 to form a second solution;

[0131] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:1 and mixed, and allowed to stand for 20 minutes to prepare a hydrogel. In this embodiment, a covalent cross-linked structure formed by four-arm polyethylene glycol maleimide and four-arm polyethylene glycol terminated with thiol groups exists inside the hydrogel.

[0132] Comparative Example 3

[0133] The preparation method of this comparative example is basically the same as that of Example 4, except that the hydrogel prepared in this comparative example only has a covalent cross-linked structure. The specific steps are as follows:

[0134] 1) Dissolve 3 g of four-arm polyethylene glycol methacrylate with a number average molecular weight of 10 kDa in 20 mL of dilute hydrochloric acid with a pH of 10 (the molar concentration is 10 -6 mol / L), forming a first solution;

[0135] 2) dissolving 3 g of amino-terminated four-arm polyethylene glycol with a number average molecular weight of 10 kDa in 20 mL of phosphate buffer at pH 9 to form a second solution;

[0136] 3) The first solution prepared in step 1) and the second solution prepared in step 2) were placed in a plastic culture dish at a volume ratio of 0.5:1 and mixed, and allowed to stand for 40 minutes to obtain a hydrogel. In this embodiment, a covalent cross-linked structure formed by four-arm polyethylene glycol methacrylate and amino-terminated four-arm polyethylene glycol exists inside the hydrogel.

[0137] Comparative Example 4

[0138] The preparation method of this comparative example is basically the same as that of Example 5, except that the hydrogel prepared in this comparative example only has a covalent cross-linked structure. The specific steps are as follows:

[0139] 1) dissolving 5 g of four-arm polyethylene glycol succinimidyl glutarate having a number average molecular weight of 10 kDa in 20 mL of dilute hydrochloric acid having a pH of 3 (the molar concentration is 0.01 mol / L) to form a first solution;

[0140] 2) dissolving 5 g of amino-terminated four-arm polyethylene glycol with a number average molecular weight of 10 kDa in 20 mL of phosphate-sodium hydroxide buffer with a pH of 9 to form a second solution;

[0141] 3) The first solution prepared in step 1) and the second solution prepared in step 2) are placed in a plastic culture dish at a volume ratio of 1:1, mixed, and allowed to stand for 0.5 minutes to prepare a hydrogel. In this embodiment, a covalent cross-linked structure formed by four-arm polyethylene glycol succinimidyl glutarate and amino-terminated four-arm polyethylene glycol exists inside the hydrogel.

[0142] Comparative Example 5

[0143] The sealant used in this comparative example is a commercially available brand Medical sealant, specification is 4mL.

[0144] Comparative Example 6

[0145] The preparation method of this comparative example is basically the same as that of the embodiment, except that the hydrogel prepared in this comparative example only has an electrostatic cross-linking structure. The specific steps are as follows:

[0146] 1) dissolving 0.04 g of carboxymethyl dextran having a number average molecular weight of 80 kDa in 20 mL of dilute hydrochloric acid having a pH of 6 to form a first solution;

[0147] 2) dissolving 0.05 g of poly-lysine with a number average molecular weight of 3 kDa in 20 mL of a phosphate buffer with a pH of 9 to form a second solution;

[0148] 3) The first solution prepared in step 1) and the second solution prepared in step 2) were placed in a plastic culture dish at a volume ratio of 0.5:1 and mixed. A white viscous precipitate was immediately observed, which was a hydrogel formed by electrostatic interaction between carboxymethyl dextran and polylysine. Due to the difficulty in regulation and characterization, performance measurement could not be performed and there was no potential application value.

[0149] The parameters of the raw materials and the like in the preparation methods of Examples 1 to 7 and Comparative Examples 1 to 5 are shown in Table 1:

[0150] Table 1

[0151]

[0152]

[0153] Performance Test:

[0154] 1. The hydrogels prepared in Examples 1 to 7 and Comparative Examples 1 to 5 were tested for swelling properties. The test results are shown in Table 2 below.

[0155] The test steps are as follows:

[0156] According to the preparation methods in Examples 2 to 7 and Comparative Examples 1 to 5, 12 cylindrical hydrogels with a diameter of about 8 mm and a height of about 5 mm were prepared using a 2 mL plastic centrifuge tube as a mold. The prepared hydrogels were then immersed in pure water at 37° C. for 12 hours, and the average swelling rate was the average growth rate of the mass of the hydrogel before and after immersion.

[0157] Table 2

[0158]

[0159]

[0160] As can be seen from Table 2 above, the present application can significantly reduce the swelling degree of the hydrogel by designing it into a multiple cross-linked network structure, thereby solving the problem that it will compress blood vessels and surrounding tissues and organs when used as a sealant. In addition, it can be seen from Examples 5, 7 and Comparative Example 4 that when there is an electrostatic cross-linked network structure in the hydrogel and the concentration of the components constituting the electrostatic cross-linked network increases, the swelling rate of the formed gel is lower, which is lower than the swelling rate of the commercial medical sealant, that is, Comparative Example 5. Moreover, the method provided by the present application can reduce the swelling rate of the hydrogel to 40.23%.

[0161] 2. The adhesion strength of the hydrogels prepared in Examples 5, 7 and Comparative Examples 4 to 5 was tested. The test results are shown in Table 3 below.

[0162] The devices used in the test process are as follows: Figure 2 The test steps are as follows:

[0163] A cleaned pigskin is taken, and a hole with a diameter of 3 mm is opened at the center thereof, and then 200 μL of the hydrogel prepared in Examples 5, 7 and Comparative Examples 4-5 are injected into the hole to block the hole. The pigskin is then fixed at one port of a three-way valve, and the other two ports of the three-way valve are respectively connected to a pressure gauge and a pipe with water, and pressurized until the hydrogel at the port is broken by the water body, and the maximum value of the pressure gauge is recorded, which is the bursting pressure before the hydrogel swells, and 5 parallel samples are set, and the average is the average bursting pressure before the hydrogel swells.

[0164] Considering the actual environment of the human body, the pigskin sealed with hydrogel was immersed in phosphate buffer solution (PBS buffer) at 37°C for 12 hours to allow the hydrogel to reach swelling equilibrium. The pigskin was taken out and fixed at the port of the three-way valve. The bursting pressure was tested again, which was the bursting pressure after the hydrogel swelled. Five parallel samples were set, and the average was the average bursting pressure after the hydrogel swelled.

[0165] The test results are shown in Table 3.

[0166] Table 3

[0167]

[0168] As can be seen from the test results in Table 3, the hydrogel prepared in this embodiment has a higher burst pressure than the hydrogel prepared in the comparative example, indicating that the hydrogel provided in this application has a higher blocking effect and is more conducive to blood vessel blocking when used as a sealant. Comparing Examples 5 and 7 with Comparative Example 4, increasing the concentration of the components constituting the electrostatic cross-linked network in the hydrogel also improves the blocking performance of the hydrogel. In addition, the burst pressures of the above three groups of samples all exceed the commercial sealant, namely Comparative Example 5, proving that they have good sealing effects.

[0169] 3. Test the gelling properties of the hydrogels obtained in Examples 2 to 5 and Comparative Examples 1 to 4. The test steps and results are as follows. Using a double injection process, the first solution and the second solution obtained in Examples 2 to 5 and Comparative Examples 1 to 4 were respectively loaded into a double syringe, extruded and mixed. During the extrusion process, it was found that after the first solution and the second solution in Examples 2 to 5 were extruded and mixed, the viscosity increased rapidly within 2 seconds; while after the first solution and the second solution in Comparative Examples 1 to 4 were extruded and mixed, the viscosity increased only after 10 seconds. It can be seen that the hydrogel provided in the present application has higher adhesion and can be better fixed to a specific position on biological tissue.

[0170] 4. Test the biocompatibility of the hydrogels prepared in Examples 5, 7 and Comparative Example 4.

[0171] 1) Cell activity test: The culture medium is Dulbecco's modified eagle medium (DMEM), the cell type is commercial porcine aortic endothelial cells, and the culture conditions are 37°C and a cell culture incubator with a carbon dioxide concentration of 5%. The hydrogel extract was prepared at a ratio of 200 mg:1 mL of hydrogel:DMEM cell culture medium, and the extract and vascular endothelial cells were co-cultured for 48 hours. The culture well plate was a 96-well plate, and 10,000 cells were added to each well in the initial stage. A blank group was set, and the cell activity of the blank group was 100%. During the experiment, all cells were cultured only with DMEM for 48 hours. The cytotoxicity after co-culture was determined using the Cell Counting Kit-8 (CCK-8) method, and the test solution was prepared at a volume ratio of CCK-8 reagent:DMEM culture medium of 1:9. Before the test, the original extract or culture medium was removed, the test solution was added, and the cells were cultured in an incubator for 2 hours. The test solution in each well was aspirated and transferred to another clean well plate. The test was performed using an ELISA instrument to measure the absorbance of the test solution in different wells at a wavelength of 450 nm. The average cell activity corresponding to Examples 5 and 7 and Comparative Example 4 and Comparative Example 4 was calculated based on the cell activity of the blank group as shown in Table 4.

[0172] Table 4

[0173] Group Average cell activity (%) Blank Group 100 Example 5 99.76 Example 7 96.28 Comparative Example 4 98.46

[0174] From the test results in Table 4, it can be seen that the hydrogel provided by the present application has excellent cell activity and can be used as a medical sealant to prevent bleeding in various parts, bleeding after vascular suture, and block blood vessels. Comparing the embodiments and comparative examples in Table 4, since the components constituting the electrostatic cross-linking network have good biocompatibility, no obvious cytotoxicity is shown after the concentration of the relevant components is increased in Example 7.

[0175] 2) Blood compatibility test: Commercially available anticoagulant sheep (anticoagulant is sodium citrate) whole blood was used for the experiment. Before use, the sheep whole blood was diluted at a volume ratio of 1:9 between sheep whole blood and phosphate buffered saline (PBS). The hydrogels prepared in Examples 1 to 7 and Comparative Examples 3 and 4 were added to the diluted sheep whole blood at a ratio of 0.1 g:1 mL of hydrogel: diluted blood, and incubated at 37°C for 1 hour. After the incubation, the hydrogel was removed, the remaining liquid was centrifuged at 1000 rpm for 10 minutes, and 100 μL of the supernatant was taken into a 96-well plate.

[0176] In addition, a positive group and a negative group were set up. Among them, the positive group was whole blood diluted with sheep whole blood and pure water at a volume ratio of 1:9, and the negative group was whole blood diluted with PBS. The negative group and the positive group were placed at 37°C for 1 hour. After the incubation, the remaining liquid was centrifuged at 1000rpm for 10 minutes, and 100μL of supernatant was taken into a 96-well plate. The embodiment and the comparative example were then tested using an ELISA instrument to measure the absorbance value of the liquid in different wells at a wavelength of 540nm. The hemolysis rate is calculated as follows:

[0177] Hemolysis rate (%) = (absorption value of sample group - absorption value of negative group) / (absorption value of positive group - absorption value of negative group) × 100%, take the average value of each group of results, the test results are shown in Table 5.

[0178] Table 5

[0179] Group Hemolysis rate (%) Negative group 0 Positive group 100 Example 1 0.25 Example 2 0.86 Example 3 0.58 Example 4 0.45 Example 5 0.31 Example 6 1.31 Example 7 0.47 Comparative Example 3 0.51 Comparative Example 4 0.36

[0180] From the test results in Table 5, it can be seen that the hydrogel provided by the present application has good blood compatibility and can be used as a sealant to prevent bleeding in various parts, bleeding after vascular suture, and block blood vessels. In addition, since the components constituting the electrostatic cross-linking network have good biocompatibility, adding them to the hydrogel also has good blood compatibility.

[0181] 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.

[0182] 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 specification and drawings may be used to interpret the scope of the claims.

Claims

1. A biological tissue adhesive hydrogel, characterized in that: include: A first component, a second component, a third component, and a fourth component; The first component and the second component form an electrostatic interaction, and the third component and the fourth component form a covalent cross-linking structure; The first component, the second component, the third component and the fourth component are all biocompatible components.

2. The hydrogel according to claim 1, characterized in that The first component includes a positively charged group, and the positively charged group includes one or more of a primary amine group, a secondary amine group, a tertiary amine group, a quaternary ammonium group, an imidazole group, a guanidine group, and a pyridine group; The second component includes negatively charged groups, and the negatively charged groups include one or more of hydroxyl groups, carboxyl groups, sulfonic acid groups and phosphate groups; The electrostatic interaction is formed between the positively charged group and the negatively charged group.

3. The hydrogel according to claim 2, characterized in that The first component includes one or more of polyethyleneimine, polyallylamine, polyethylene glycol modified with the positively charged group, polysaccharide containing or modified with the positively charged group, and polyamino acid containing or modified with the positively charged group; The second component includes one or more of polyacrylic acid and its water-soluble derivatives, polyethylene glycol modified with the negatively charged group, polysaccharide containing or modified with the negatively charged group, and polyamino acid containing or modified with the negatively charged group.

4. The hydrogel according to claim 1, wherein The third component includes a first reactive group, wherein the first reactive group includes one or more of a hydroxyl group, a thiol group, an amino group, an imino group and an azide group; The fourth component includes a second reactive group, and the second reactive group includes one or more of anhydride, aldehyde, succinimide, maleimide, carbon-carbon double bond, carbon-carbon triple bond, isocyanate, epoxy, carboxyl and isothiocyanate; The first reactive group and the second reactive group can be chemically cross-linked.

5. The hydrogel according to claim 4, characterized in that The third component includes one or more of polyethyleneimine, polyamino acid, water-soluble protein, gelatin and polyethylene glycol modified with the first active group; The fourth component includes one or both of polyethylene glycol modified with the second active group and polysaccharide modified with the second active group.

6. The hydrogel according to any one of claims 1 to 5, characterized in that The number average molecular weights of the first component and the second component are independently 1 kDa to 2×10 3 kDa, the number average molecular weight of the third component and the fourth component are independently 100Da to 2×10 3 kDa.

7. The hydrogel according to any one of claims 1 to 5, characterized in that The ratio of the sum of the mass of the first component and the second component to the sum of the mass of the third component and the fourth component is 1:(1-1000); and / or, the mass ratio of the first component to the second component is 1:(0.05-20); And / or, the mass ratio of the third component to the fourth component is 1:(0.02-50).

8. A method for preparing a hydrogel according to any one of claims 1 to 7, characterized in that: The following steps are involved: dissolving the first component and the third component in a first solvent to form a first solution; dissolving the second component and the fourth component in a second solvent to form a second solution; The first solution and the second solution are mixed so that the first component and the second component form a non-covalent cross-linked network, and the third component and the fourth component form a covalent cross-linked network, to prepare the hydrogel.

9. The preparation method according to claim 8, characterized in that: Meet at least one of the following characteristics: 1) In the first solution, the concentration of the first component is 0.05 w / v% to 1 w / v%, and the concentration of the third component is 1 w / v% to 50 w / v%; 2) In the second solution, the concentration of the second component is 0.05 w / v% to 1 w / v%, and the concentration of the fourth component is 1 w / v% to 50 w / v%; 3) The volume ratio of the first solution to the second solution is (0.01-1):(1-0.01).

10. The preparation method according to claim 8, characterized in that: The pH value of the first solvent is 1≤pH<7, and the pH value of the second solvent is 7≤pH≤13.

11. The preparation method according to any one of claims 8 to 10, characterized in that: The first solvent comprises one or more of hydrochloric acid solution, sulfuric acid solution, phosphoric acid solution, phosphoric acid-phosphate buffer, citric acid-citrate buffer, boric acid-borate buffer, acetic acid-acetate buffer, glycine-hydrochloric acid buffer, amino acid buffer and amino acid salt buffer; And / or, the second solvent includes one or more of pure water, physiological saline, sodium hydroxide solution, potassium hydroxide solution, phosphate buffer, carbonate buffer, borate buffer, acetate buffer, citrate buffer, glycine-sodium hydroxide buffer, amino acid buffer and amino acid salt buffer solution.

12. Use of the hydrogel according to any one of claims 1 to 7 in the preparation of medical sealants or as in vitro medical sealants.

Images