Method for preparing bioprosthetic valve material through double-bond polymerization after aldehyde group crosslinking, bioprosthetic valve material and application of bioprosthetic valve material

Abstract

The invention discloses a method for preparing a biological valve material through double-bond polymerization after aldehyde group crosslinking, the biological valve material and application. The preparation method comprises the following steps: step S110, enabling a biological material to be in contact with an aldehyde group crosslinking agent solution for crosslinking; s120, soaking the biological material treated in the step S110 in a solution containing a first functional monomer, and reacting to access a first carbon-carbon double bond; the first functional monomer has a first carbon-carbon double bond and an oxirane group; and step S200, carrying out polymerization reaction on the carbon-carbon double bonds under the action of an initiator to obtain the bioprosthetic valve material. After double bonds are introduced to the glutaraldehyde cross-linked bioprosthetic valve material, polymerization of the double bonds is further initiated, the stability of the glutaraldehyde cross-linked material is improved, and the calcification risk caused by structural degradation is further reduced, so that the bioprosthetic valve material also has certain anti-calcification ability.

Description

Method for preparing biological valve material by double bond polymerization after aldehyde cross-linking, biological valve material and application thereof Technical Field

[0001] The present application relates to the technical field of interventional materials, and in particular to a method for preparing a biological valve material by double bond polymerization after aldehyde cross-linking, the biological valve material, and its application. Background Art

[0002] Biological heart valves are usually made from pig or cow pericardium and are used to replace functionally defective human heart valves. Biological heart valves have many advantages over mechanical heart valves: patients do not need to take anticoagulants for a long time after implantation of biological heart valves, and biological heart valves can be implanted using minimally invasive surgical methods. These advantages have made biological heart valves gradually become the mainstream market in clinical applications.

[0003] Almost all bioprosthetic valves currently on the market are cross-linked using glutaraldehyde. While glutaraldehyde can improve the mechanical properties of the pericardium and reduce its immunogenicity to a certain extent, the stability and degree of cross-linking of glutaraldehyde-crosslinked bioprosthetic valves remain low. This can lead to component degradation after implantation, disrupting their structure and causing structural degradation. Furthermore, the degradation of bioprosthetic valve components can further induce mechanical damage and calcification, compromising normal valve function and shortening their service life.

[0004] Glutaraldehyde cross-linking is still the mainstream method for current bioprosthetic valve products. Therefore, further modification of bioprosthetic valves based on glutaraldehyde cross-linking to improve their cross-linking degree and stability is of great significance to scientific research and the development of related industrial fields.

[0005] The applicant of the present application has long been committed to the research of biological heart valves. For example, in previous research, the Chinese invention patent application document with publication number CN 114748694A disclosed a co-cross-linked biological valve material and its preparation method and application, in which the biological valve material was functionally modified by introducing functional monomers for co-cross-linking during the cross-linking treatment; in the biological valve preparation methods disclosed in the Chinese invention patent application documents with publication numbers CN 114748693A, CN114748697A, CN 114748696A and CN 114748695A, while adding functional monomers for co-cross-linking, carbon-carbon double bonds were introduced from the functional monomers as a basis for further cross-linking, and the modification of the biological valve material was completed through two cross-linking processes.

[0006] In the studies mentioned above, whether it is the introduction of functional monomers for co-crosslinking modification during glutaraldehyde crosslinking, or the introduction of carbon-carbon double bonds as the basis for further crosslinking during the co-crosslinking process, new modified substances are introduced into the glutaraldehyde crosslinking process to participate in the crosslinking reaction.

[0007] Summary of the Invention

[0008] The present application provides a method for preparing a biological valve material by double bond polymerization after aldehyde cross-linking, as well as a biological valve material and its application. After glutaraldehyde cross-linking, functional monomers with carbon-carbon double bonds are introduced from the active groups on the glutaraldehyde cross-linked membrane, such as residual amino groups, hydroxyl groups, and carboxyl groups, thereby providing a controllable cross-linking opportunity and range for the glutaraldehyde cross-linked membrane.

[0009] A method for preparing a biological valve material by double bond polymerization followed by aldehyde cross-linking, comprising:

[0010] Step S110: contacting the biomaterial with an aldehyde cross-linking agent solution for cross-linking;

[0011] Step S120: Soaking the biomaterial treated in step S110 in a solution containing a first functional monomer to chemically connect the first carbon-carbon double bond; the first functional monomer has a first carbon-carbon double bond and an ethylene oxide group;

[0012] In step S200 , a carbon-carbon double bond is polymerized under the action of an initiator to obtain a biological valve material.

[0013] Optionally, the aldehyde-based cross-linking agent is glutaraldehyde or formaldehyde.

[0014] Optionally, the biological material is animal tissue, including one or more of pericardium, valve, intestinal membrane, meninges, lung membrane, blood vessel, skin or ligament.

[0015] Optionally, the animal tissue is fresh animal tissue or biological tissue that has been decellularized.

[0016] In step S200 : an initiator is added to the system processed in the previous step; or the biological valve material processed in the previous step is washed and then immersed in a solution containing the initiator.

[0017] Optionally, the initiator is a single initiator or a mixed initiator.

[0018] Optionally, the mixed initiator is:

[0019] A mixture of ammonium persulfate and sodium bisulfite, or a mixture of ammonium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium sulfite, or a mixture of potassium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium bisulfite, or a mixture of potassium persulfate and sodium bisulfite, or potassium persulfate and tetramethylethylenediamine, or ammonium persulfate and tetramethylethylenediamine, or sodium persulfate and tetramethylethylenediamine; the concentration of each component in the mixture is 1 to 100 mM.

[0020] Optionally, the single initiator is any component of the mixed initiators.

[0021] Optionally, in step S200, the double bond polymerization time is 3 to 24 hours.

[0022] Optionally, the first functional monomer is selected from at least one of allyl glycidyl ether, glycidyl methacrylate and glycidyl acrylate.

[0023] Optionally, in step S110:

[0024] The w / w concentration of the aldehyde cross-linking agent solution is 0.1% to 5%; and the cross-linking time is 0.5h to 120h.

[0025] Optionally, in step S120:

[0026] The w / w concentration of the first functional monomer in the solution containing the first functional monomer is 1% to 10%; and the reaction time is 2 to 120 hours.

[0027] Optionally, the solution containing the first functional monomer only contains the first functional monomer and a solvent that does not participate in the chemical reaction.

[0028] Optionally, the solvent in the solution containing the first functional monomer is one or more of an aqueous solution of any one of methanol, ethanol, ethylene glycol, propanol, 1,2-propylene glycol, 1,3-propylene glycol, isopropanol, butanol, isobutanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol and glycerol, water, physiological saline, and pH neutral buffer.

[0029] The present application also provides a biological valve material prepared by the above-mentioned preparation method.

[0030] The present application also provides a biological valve material, comprising:

[0031] Step S110: contacting the biomaterial with an aldehyde cross-linking agent solution for cross-linking;

[0032] Step S120: Soaking the biomaterial treated in step S110 in a solution containing a first functional monomer to chemically connect the first carbon-carbon double bond; the first functional monomer has a first carbon-carbon double bond and an ethylene oxide group;

[0033] In step S200 , a carbon-carbon double bond is polymerized under the action of an initiator to obtain a biological valve material.

[0034] The present application also provides a biological valve, comprising a stent and leaflets, wherein the leaflets are made of the biological valve material.

[0035] Optionally, the biological valve is a heart valve.

[0036] The present application also provides an interventional system, comprising a heart valve and a catheter assembly, wherein the heart valve is folded and transported by the catheter assembly, and the heart valve comprises a stent and leaflets, wherein the leaflets are made of the biological valve material.

[0037] Compared with the prior art, this application has at least one of the following beneficial effects:

[0038] (1) The method of the present application introduces double bonds as the basis for secondary cross-linking on the biological valve material cross-linked with glutaraldehyde through double bond modification, and further achieves secondary cross-linking by initiating polymerization of double bonds on the biological valve material cross-linked with glutaraldehyde, which can further increase the cross-linking degree of the biological valve material and thus improve the stability of the biological valve material.

[0039] (2) The present application introduces double bonds into the glutaraldehyde cross-linked biological valve material, further initiating the polymerization of the double bonds, thereby improving the stability of the glutaraldehyde cross-linked material and further reducing the risk of calcification caused by structural degradation. Therefore, the material also has certain anti-calcification properties.

[0040] (3) Compared with the modification method of the applicant's previous research on the glutaraldehyde modification process in which carbon-carbon double bonds are introduced by adding functional monomers for co-crosslinking, in the modification process of the bioprosthetic valve material of the present application, glutaraldehyde crosslinking treatment is first performed, and then the residual amino groups and active groups such as hydroxyl and carboxyl groups on the glutaraldehyde crosslinking membrane are chemically connected to the functional monomers with carbon-carbon double bonds. The functional monomers with carbon-carbon double bonds are chemically connected with the amino groups, hydroxyl groups and carboxyl groups on the surface of the glutaraldehyde crosslinking membrane through the ethylene oxide group, and the carbon-carbon double bonds are mainly connected to the surface of the bioprosthetic valve material. In the process of glutaraldehyde crosslinking modification of the bioprosthetic valve material, no other substances that can participate in the crosslinking reaction are added, which can better protect the original fiber structure of the biomaterial, and can effectively ensure the mechanical properties of the membrane while ensuring the orientation direction of the original fibers of the biomaterial, avoiding the problem in the previous research that the direct addition of double-bond functional monomers during crosslinking may destroy the original fiber orientation of the biomaterial and increase the fiber disorder. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] FIG1 is a process flow chart of a preferred embodiment of the present application;

[0042] FIG2 is a reaction principle diagram of a preferred embodiment of the present application;

[0043] FIG3 is an Alizarin red staining result of control group 1 (glutaraldehyde cross-linked porcine pericardium) 30 days after subcutaneous implantation in rats;

[0044] FIG4 is an Alizarin red staining result of Sample 1 of Example 1 after subcutaneous implantation in rats for 30 days;

[0045] FIG5 is an Alizarin red staining result of Sample 5 of Example 5 after subcutaneous implantation in rats for 30 days;

[0046] FIG6 is an Alizarin red staining result of Sample 7 of Example 7 after subcutaneous implantation in rats for 30 days;

[0047] FIG7 is a schematic structural diagram of the heart valve of the present application;

[0048] FIG8 is a schematic structural diagram of the intervention system of the present application. DETAILED DESCRIPTION

[0049] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0050] 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 pertains. The terms used herein in the specification of this application are for the purpose of describing specific embodiments only and are not intended to limit this application.

[0051] Currently, almost all biological valve products on the market are made by cross-linking with glutaraldehyde. Glutaraldehyde can improve the mechanical properties of the pericardium and reduce its immunogenicity to a certain extent. However, the stability and cross-linking degree of glutaraldehyde-cross-linked biological valves are still relatively low, which will lead to the degradation of its components after implantation, causing its structure to be destroyed and structural degradation to occur. Furthermore, the degradation of biological valve components will further induce mechanical damage and calcification, affecting the normal function of the valve and reducing its service life. Glutaraldehyde cross-linking is still the mainstream method for current biological valve products. Therefore, further cross-linking and modification of biological valves on the basis of glutaraldehyde cross-linking to improve their cross-linking degree and stability is of great significance to scientific research and the development of related industries.

[0052] This application further introduces double bonds and initiates post-crosslinking on the basis of glutaraldehyde crosslinking, that is, on the basis of the glutaraldehyde crosslinked membrane, the first carbon-carbon double bond is introduced by chemically bonding the first functional monomer (containing a first carbon-carbon double bond and an ethylene oxide group) to the glutaraldehyde crosslinked biological membrane. This will improve the crosslinking degree, stability, mechanical properties and anti-calcification of the glutaraldehyde crosslinked biological valve material membrane.

[0053] In one embodiment, it specifically includes (see Figure 1):

[0054] S110 immersing the biological valve material in an aldehyde-based crosslinking agent solution for crosslinking to prepare a glutaraldehyde-crosslinked biological valve material;

[0055] S120: Soaking the glutaraldehyde-crosslinked bioprosthetic valve material prepared in step S110 in a solution containing a double-bonding agent (first functional monomer) for double-bonding modification to prepare a double-bonded bioprosthetic valve material; the double-bonding agent (first functional monomer) has at least one first carbon-carbon double bond and an ethylene oxide group.

[0056] S200 brings the bioprosthetic valve material treated in step S120 into contact with an initiator to initiate double bond polymerization.

[0057] In the present application, the biomaterial is first subjected to a cross-linking reaction with an aldehyde cross-linking agent (S110), and then reacts with the active group of the first functional monomer to access the first carbon-carbon double bond (S120). During the preparation process, an aldehyde cross-linking agent is first added, and the aldehyde cross-linking agent first reacts with part of the amino group of the biomaterial, and then the first functional monomer is added, and the remaining amino groups and other groups (such as hydroxyl and carboxyl) on the biomaterial are used to react with the active groups on the first functional group to directly access the first carbon-carbon double bond. In this scheme, the first functional monomer also carries an oxirane group as an active group, which participates in the chemical reaction through the active group. In addition to the remaining amino groups on the biomaterial participating in the reaction, its hydroxyl and carboxyl groups can also react with the oxirane group and participate in the chemical reaction. The first carbon-carbon double bond introduced by the chemical reaction is then polymerized under the action of an initiator to further form a cross-linked network, thereby improving the anti-coagulation, anti-calcification, elasticity and other properties of the biological valve cross-linked based on glutaraldehyde.

[0058] The reaction principle of this application:

[0059] In this double bond cross-linking scheme, after the biological valve material is cross-linked with glutaraldehyde, the double bond of the glutaraldehyde cross-linked biological valve material is further achieved by introducing a first carbon-carbon double bond using a first functional monomer, i.e., a double bond-forming agent. The first functional monomer, i.e., the double bond-forming agent, used has both a first carbon-carbon double bond and an ethylene oxide group.

[0060] To facilitate understanding of the chemical principles involved in this scheme, the following is further illustrated using Figure 2 as an example: The glutaraldehyde-crosslinked bioprosthetic valve material is modified using the first functional monomer, a double-bonding agent. The oxirane group in the first functional monomer, a double-bonding agent, undergoes a ring-opening reaction with the hydroxyl and carboxyl groups on the glutaraldehyde-crosslinked bioprosthetic valve material, as well as the small amount of amino groups remaining after glutaraldehyde crosslinking. This directly introduces a first carbon-carbon double bond into the glutaraldehyde-crosslinked bioprosthetic valve material. Furthermore, polymerization of these double bonds on the glutaraldehyde-crosslinked bioprosthetic valve material is initiated, achieving secondary crosslinking and completing the post-crosslinking treatment of the bioprosthetic valve material. The crosslinking degree of the bioprosthetic valve material after secondary crosslinking is further enhanced, while its stability, mechanical properties, and anti-calcification properties are also further improved.

[0061] After glutaraldehyde cross-linking, carbon-carbon double bonds are introduced. The carbon-carbon double bonds are mainly connected to the surface of the biological valve material. In the process of glutaraldehyde cross-linking modification of the biological valve material, no other substances that can participate in the cross-linking reaction are added, which can better protect the original fiber structure of the biomaterial. While effectively ensuring the mechanical properties of the membrane, the orientation direction of the original fibers of the biomaterial can be guaranteed, avoiding the problem in previous studies that the direct addition of double-bond functional monomers during cross-linking may destroy the original fiber orientation of the biomaterial and increase the fiber disorder.

[0062] Optionally, in step S120 of the present application, non-condensing chemical bonding is used to connect the first carbon-carbon double bond.

[0063] Optionally, in step S110, the biomaterial is not subjected to any other chemical reaction involving reagents before being treated with the aldehyde cross-linking agent.

[0064] Further optionally, in the reaction system of step S120, the first carbon-carbon double bond is provided by a first functional monomer having an active group, and the reaction raw materials in steps S110 and S120 only include the biomaterial, the first functional monomer and the aldehyde cross-linking agent.

[0065] In step S110:

[0066] The cross-linking agent of the present application adopts the aldehyde cross-linking agent used in the current mainstream cross-linking method. Optionally, the aldehyde cross-linking agent can be selected from glutaraldehyde and formaldehyde.

[0067] Optionally, the concentration of the glutaraldehyde solution is 0.1% to 5% (w / w); and the cross-linking time can be any time between 0.5 h and 120 h.

[0068] The biomaterial used in this application is a conventional biomaterial used in existing glutaraldehyde cross-linking processes, and the collagen content of the biomaterial is 60% to 90%. Furthermore, the biomaterial is animal tissue, and the animal source is pig, cow, horse, or sheep, including one or more of the pericardium, valve, intestinal membrane, meninges, lung membrane, blood vessels, skin, or ligaments.

[0069] Optionally, the animal tissue is fresh animal tissue or biological tissue that has been decellularized.

[0070] Optionally, in the decellularization step, the biological tissue is treated with a surfactant as follows:

[0071] Decellularization of biological tissue using ionic surfactants; or

[0072] Non-ionic surfactants are used to decellularize biological tissues.

[0073] The ionic surfactant is mainly used for lysing cells, and the nonionic surfactant is mainly used for removing lipid substances (such as phospholipids).

[0074] Optionally, the ionic surfactant is at least one of sodium deoxycholate, fatty acid potassium soap, sodium lauryl sulfate, sodium cholate, hexadecyltrimethylammonium bromide, fatty acid potassium salt, and alkyldimethylsulfonpropyl betaine.

[0075] Optionally, the nonionic surfactant is at least one of Triton and Tween.

[0076] In step S120:

[0077] Optionally, the double-bonding agent, ie, the first functional monomer, is selected from at least one of allyl glycidyl ether, glycidyl methacrylate, and glycidyl acrylate.

[0078] Optionally, the concentration of the double-bonding agent in the solution containing the first functional monomer, ie, the double-bonding agent, is 1% to 10% (w / w); and the reaction time for the double-bonding modification is 2 to 120 hours.

[0079] Optionally, the solvent in the solution containing the first functional monomer, i.e., the double-bonding agent, is one or more of water, physiological saline, pH neutral buffer, or an aqueous solution of methanol, ethanol, ethylene glycol, propanol, 1,2-propylene glycol, 1,3-propylene glycol, isopropanol, butanol, isobutanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, or glycerol.

[0080] Optionally, the biofilm material treated in S110 is taken out and washed or directly placed in a solution containing a double-bonding agent (first functional monomer).

[0081] In step S200:

[0082] The bioprosthetic valve material treated in step S120 is washed with deionized water and then immersed in an initiator solution for treatment in step S200 or an initiator is directly added to the reaction system in step S120 to initiate polymerization, the latter being commonly known as a one-pot method.

[0083] Optionally, the solvent in the initiator-containing solution is water, physiological saline or pH neutral buffer.

[0084] As mentioned above, the concentration of the initiator can be understood as the concentration of the initiator in the solution contained in the reaction system in step S120 in the one-pot method, and can be understood as the concentration in the solution containing the initiator in the step-by-step method.

[0085] Optionally, the initiator is a mixture of ammonium persulfate and sodium bisulfite, or a mixture of ammonium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium sulfite, or a mixture of potassium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium bisulfite, or a mixture of potassium persulfate and sodium bisulfite, or potassium persulfate and tetramethylethylenediamine, or ammonium persulfate and tetramethylethylenediamine, or sodium persulfate and tetramethylethylenediamine; the concentration of each component in the mixture is 1 to 100 mM.

[0086] Optionally, the double bond polymerization time is preferably 3 to 24 hours.

[0087] In the present application, all reaction processes of S110, S120 and S200 can be carried out at 0-50°C unless otherwise specified. Preferably, the temperature does not need to be specially controlled and can be carried out at room temperature, preferably not exceeding the temperature adapted to the human body, and preferably at 36-37°C.

[0088] In the present application, all reactions of S110, S120 and S200 can be either static reactions or dynamic reactions unless otherwise specified. The dynamic reactions can be carried out under the action of a peristaltic pump or other equipment that can circulate the solution, or can be carried out by shaking at a speed of 10 rpm-150 rpm. The peristaltic cycle or shaking time can be continuous or intermittent.

[0089] In the present application, the double bond polymerization may be followed by dehydration and drying to produce a dry film. After the double bond polymerization, the bioprosthetic valve material may be cleaned and softened in a conventional manner and then dehydrated and dried.

[0090] The cleaning solution can be one or a mixture of water, physiological saline, ethanol, isopropanol or pH neutral buffer solution. The pH can be adjusted to between 5.0 and 9.5 before and during use, or it can be left unadjusted.

[0091] Optionally, the dehydration treatment is to expose the membrane sheet after double bond polymerization or the valve sewn from the membrane sheet to a dehydration solution.

[0092] Optionally, the dehydration solution is a mixed solution of an alcohol solution and water, the alcohol solution accounts for 20-90% (v / v), and the alcohol reagent can be one of ethanol and isopropanol or a mixture of the two.

[0093] Optionally, the drying treatment is to expose the dehydrated membrane or valve to a softener solution for a treatment time of 20 minutes to 10 hours.

[0094] Optionally, the main component of the softener solution is a mixed solution of one or two of glycerol and polyethylene glycol, the glycerol concentration is 10-100% (v / v), and the other components are one or more of water, ethanol, and isopropanol, accounting for 0-90% (v / v).

[0095] Optionally, the valve after drying can be sterilized by ethylene oxide sterilization or electron beam sterilization.

[0096] The bioprosthetic valve material prepared by the above method can be used for interventional bioprosthetic valves, such as through minimally invasive intervention; it can also be used for surgical bioprosthetic valves, such as through surgical implantation.

[0097] As shown in Figure 7, in one embodiment, an artificial heart valve is provided, including a stent 1 and leaflets 2 connected to the stent 1. The stent is cylindrical as a whole, and the side walls are a hollow grid structure. The interior of the stent is a blood flow channel, and the multiple leaflets cooperate with each other to control the degree of opening and closing of the blood flow channel in the stent.

[0098] Depending on the release mode, the stent is processed using corresponding materials, such as nickel-titanium alloy with shape memory that can self-expand in the body, or stainless steel that is released by balloon expansion, etc. The stent itself can be formed by cutting tubes or weaving wires, and the leaflets can be connected to the stent by sewing, bonding or integral mold molding.

[0099] To ensure positioning within the body, positioning structures that can interact with surrounding native tissues, such as anchors or arms, can be provided on the periphery of the stent. To prevent leakage, skirts or anti-leakage materials can be provided on the inner and / or outer sides of the stent. The leaflets, skirts, or anti-leakage materials can all be made of the bioprosthetic valve materials described in the above embodiments.

[0100] As shown in Figure 8, when catheter intervention is used, the artificial heart valve 3 and the corresponding delivery system constitute a valve intervention system. The delivery system includes a catheter assembly 4 and a handle for controlling the catheter assembly. The artificial heart valve is in a radially compressed state when delivered in the body. The catheter assembly is released from its restraints or undergoes balloon expansion and radial expansion and release in the body.

[0101] The following is further described with specific examples:

[0102] Control group 1

[0103] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100 RPM for 2 hours, then immersed in 0.30% (w / w) glutaraldehyde solution and cross-linked at room temperature and 100 RPM for 48 hours to obtain control sample 1.

[0104] Example 1

[0105] In this embodiment, freshly collected porcine pericardium was washed with distilled water at 4°C and 100 RPM for 2 hours, immersed in a 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde-cross-linked porcine pericardium.

[0106] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 5% (v / v) propanol aqueous solution of glycidyl methacrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 20% (v / v) propanol aqueous solution.

[0107] After the double bond modification is completed, the double-bonded glutaraldehyde cross-linked pig pericardium is washed with deionized water; then the double-bonded glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 20 mM potassium persulfate and 10 mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double-bonded cross-linked pig pericardium is obtained, which is recorded as sample 1.

[0108] Example 2

[0109] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0110] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 6% (v / v) isopropanol aqueous solution of glycidyl acrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 20% (v / v) isopropanol aqueous solution.

[0111] After the double bond modification is completed, the double-bond glutaraldehyde cross-linked pig pericardium is washed with deionized water; then the double-bond glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 20mM ammonium persulfate and 5mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double-bond post-cross-linked pig pericardium is obtained, which is recorded as sample 2.

[0112] Example 3

[0113] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0114] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in an isopropanol aqueous solution containing 4% (v / v) glycidyl acrylate and 4% (v / v) allyl glycidyl ether at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was 30% (v / v) ethanol aqueous solution.

[0115] After the double bond modification was completed, the double-bonded glutaraldehyde cross-linked pig pericardium was washed with deionized water; then the double-bonded glutaraldehyde cross-linked pig pericardium was immersed in a mixture of 20 mM ammonium persulfate and 10 mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 7 hours, the double-bonded cross-linked pig pericardium was obtained, which was recorded as sample 3 and numbered GAGA-PP-3.

[0116] Example 4

[0117] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0118] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in an isopropanol aqueous solution of 5% (v / v) glycidyl methacrylate and 2% (v / v) glycidyl acrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 35% (v / v) isopropanol aqueous solution.

[0119] After the double bond modification was completed, the double bond glutaraldehyde cross-linked pig pericardium was washed with deionized water; then the double bond glutaraldehyde cross-linked pig pericardium was immersed in a mixture of 20 mM sodium persulfate and 5 mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double bond post-cross-linked pig pericardium was obtained, which was recorded as sample 4.

[0120] Example 5

[0121] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0122] The glutaraldehyde-crosslinked porcine pericardium was washed with deionized water and immersed in a 4% (v / v) ethanol aqueous solution of glycidyl methacrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution was 20% (v / v) ethanol aqueous solution.

[0123] After the double bond modification was completed, ammonium persulfate and sodium bisulfite were added to initiate the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium, wherein the ammonium persulfate concentration was 20 mM and the sodium bisulfite concentration was 5 mM; after adding the initiator and reacting at 37°C for 8 hours, the double-bond cross-linked pig pericardium was obtained, which was recorded as sample 5.

[0124] Example 6

[0125] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0126] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 4% (v / v) isobutanol aqueous solution of glycidyl methacrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 15% (v / v) isobutanol aqueous solution.

[0127] After the double bond modification was completed, the double bond glutaraldehyde cross-linked pig pericardium was washed with deionized water; then the double bond glutaraldehyde cross-linked pig pericardium was immersed in a mixture of 20 mM ammonium persulfate and 5 mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double bond post-cross-linked pig pericardium was obtained, which was recorded as sample 6.

[0128] Example 7

[0129] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0130] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 4% (v / v) isopropanol aqueous solution of glycidyl acrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 48 hours, and the solvent of the double bond modification solution used was a 20% (v / v) methanol aqueous solution.

[0131] After the double bond modification was completed, the double bond glutaraldehyde cross-linked pig pericardium was washed with deionized water; then the double bond glutaraldehyde cross-linked pig pericardium was immersed in a mixture of 20 mM ammonium persulfate and 6.5 mM sodium sulfite to further induce the polymerization reaction of the double bonds on the double bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 10 hours, the double bond post-cross-linked pig pericardium was obtained, which was recorded as sample 7.

[0132] Example 8

[0133] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0134] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 4% (v / v) aqueous solution of glycidyl methacrylate in ethylene glycol at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 20% (v / v) aqueous solution of ethylene glycol.

[0135] After the double bond modification is completed, the double-bond glutaraldehyde cross-linked pig pericardium is washed with deionized water; then the double-bond glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 40mM ammonium persulfate and 15mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 7 hours, the double-bond post-cross-linked pig pericardium is obtained, which is recorded as sample 8.

[0136] Example 9

[0137] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0138] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 7% (v / v) propanol aqueous solution of glycidyl acrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 60 hours, and the solvent of the double bond modification solution used was 40% (v / v) propanol aqueous solution.

[0139] After the double bond modification is completed, the double-bonded glutaraldehyde cross-linked pig pericardium is washed with deionized water; then the double-bonded glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 30mM sodium persulfate and 10mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double-bonded cross-linked pig pericardium is obtained, which is recorded as sample 9.

[0140] Example 10

[0141] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0142] After washing with deionized water, the glutaraldehyde-crosslinked porcine pericardium was immersed in an isopropanol aqueous solution containing 6% (v / v) glycidyl methacrylate and 3% (v / v) glycidyl acrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 84 hours, and the solvent of the double bond modification solution used was 50% (v / v) ethanol aqueous solution.

[0143] After the double bond modification is completed, the double-bonded glutaraldehyde cross-linked pig pericardium is washed with deionized water; then, the double-bonded glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 40 mM ammonium persulfate and 10 mM sodium sulfite to further induce the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 12 hours, the double-bonded cross-linked pig pericardium is obtained, which is recorded as sample 10.

[0144] The performance of the samples of Examples 1 to 10 and Control Group 1 was characterized:

[0145] To characterize the changes in the degree of cross-linking of glutaraldehyde-cross-linked bioprosthetic valve materials before and after double-bond post-cross-linking treatment, the thermal stability and cross-linking degree of the bioprosthetic valve materials were characterized by measuring the thermal shrinkage temperature of the bioprosthetic valve materials; the stability of the bioprosthetic valve materials was characterized by an enzyme degradation experiment; and the calcification degree (anti-calcification performance) of the samples was characterized by a rat subcutaneous implantation experiment.

[0146] Thermal shrinkage temperature determination:

[0147] The bioprosthetic valve material was cut into circular sheets with a diameter of 0.6 cm, dried, and placed in a crucible. The thermal shrinkage temperature of the bioprosthetic valve material was measured on a differential scanning calorimeter at a heating rate of 10°C / min in the range of 40-120°C. The thermal shrinkage temperature was measured to characterize the thermal stability and degree of cross-linking of the bioprosthetic valve material; higher thermal shrinkage temperatures correspond to higher thermal stability and degree of cross-linking.

[0148] Table 1 Heat shrinkage temperature of each group of samples

[0149] Sample thermal shrinkage temperature (°C) Control group 1 (glutaraldehyde cross-linked pig pericardium) 84.7 Example 1 88.9 Example 2 89.3 Example 9 91.5 Example 10 92.0

[0150] Thermal shrinkage temperature measurements were performed on Examples 1, 2, 9, 10, and Control Group 1 (glutaraldehyde-crosslinked porcine pericardium). As shown in Table 1, the thermal shrinkage temperatures of Examples 1, 2, 9, and 10 were all higher than those of Control Group 1 (glutaraldehyde-crosslinked porcine pericardium). This means that the thermal stability and degree of crosslinking of Examples 1, 2, 9, and 10 were all higher than those of Control Group 1 (glutaraldehyde-crosslinked porcine pericardium). The results of the thermal shrinkage temperature measurement experiments demonstrate that the present method for preparing bioprosthetic valve materials by double-bond post-crosslinking can improve the thermal stability and degree of crosslinking of bioprosthetic valves.

[0151] Enzyme degradation experiment

[0152] Samples 3, 6, 10, and control group 1 were cut into 1-cm-diameter circular sheets, with six replicates per group. All circular sheets were placed in a 48-well plate, frozen overnight at -80°C, and then transferred to a vacuum freeze dryer for 48 hours. Each sheet was weighed on a 1 / 100,000 balance, recorded as the initial weight (W0), and returned to the 48-well plate. Using a pipette, 0.5 mL of collagenase I in PBS was added to each well of the 48-well plate, ensuring that the bioprosthetic valve sample was completely immersed in the collagenase solution (100 U / mL). The 48-well plate was then transferred to a 37°C incubator and incubated for 24 hours. After incubation, the solution in the plate was discarded, and deionized water was pipetted into the bioprosthetic valve sample repeatedly with a rubber-tipped pipette. After three washes, the sample was frozen overnight at -80°C and then transferred to a vacuum freeze dryer for 48 hours. The weight of each sample after collagenase degradation was weighed on a 1 / 100,000 balance and recorded as the final weight (Wt). The formula for calculating the weight loss rate due to enzyme degradation is as follows:

[0153]

[0154] The results are shown in Table 2:

[0155] Table 2 Enzyme degradation weight loss rate of each group of samples

[0156] Sample enzymatic degradation weight loss rate (%) Control group 1 (glutaraldehyde cross-linked pig pericardium) 7.45±1.33 Sample 3 5.31±0.30 Sample 4 4.47±1.05 Sample 6 5.12±0.97

[0157] Sample 103.06±0.59

[0158] Enzyme degradation experiments were performed on the control group 1 (glutaraldehyde cross-linked porcine pericardium), sample 3, sample 4, sample 6, and sample 10 to characterize the cross-linking efficiency of each group of samples. After treating the control group 1 (glutaraldehyde cross-linked porcine pericardium), sample 3, sample 4, sample 6, and sample 10 with collagenase I, the enzyme degradation weight loss rate of each group of samples was calculated as shown in Table 2. The enzyme degradation weight loss rates of sample 3, sample 4, sample 6, and sample 10 were all lower than that of the control group (glutaraldehyde cross-linked porcine pericardium), which indicates that the stability of sample 3, sample 4, sample 6, and sample 10 was higher than that of the control group (glutaraldehyde cross-linked porcine pericardium), that is, sample 3, sample 4, sample 6, and sample 10 were higher. The results of the enzyme degradation experiment show that the method of preparing biological valve materials by double bond post-crosslinking of the present application can improve the stability of biological valves.

[0159] Anti-calcification test

[0160] Cut the sample bioprosthetic valve material into 0.8◇0.8cm 2 After sterilization, the samples were implanted subcutaneously in rats and removed 30 days later. Each sample was divided into two parts. One part was decapsulated, freeze-dried, weighed, and digested with 6M hydrochloric acid for determination of calcium content per gram. The other part was fixed with paraformaldehyde tissue fixative. After fixation, the samples were removed, trimmed with a scalpel, and transferred to a dehydration box. The samples were dehydrated using a gradient of ethanol. After dehydration, the samples were transferred to an embedding machine and embedded in melted paraffin wax. They were then cooled in a -20°C refrigerator and trimmed. 5μm-thick sections were cut from the trimmed wax blocks on a microtome, transferred from the slide spreader to glass slides, dewaxed, and rehydrated. The sections were stained with alizarin red for 3 minutes, washed with water, dried, and permeabilized with xylene for 5 minutes. The sections were mounted with neutral gum, and images of the staining results were captured on a pathology slide scanner.

[0161] Table 3 Calcium content of samples in each group after 30 days of subcutaneous implantation in rats

[0162] Calcium content of samples (mg / g) Control group 1 (glutaraldehyde cross-linked pig pericardium) 74.9±12.3 Sample 1 15.1±4.7 Sample 5 8.4±4.6 Sample 7 12.7±5.1

[0163] Calcium content was measured in Samples 1, 5, and 7, as well as in Control 1 (glutaraldehyde-crosslinked porcine pericardium), 30 days after subcutaneous implantation in rats, to characterize the degree of calcification in each group. As shown in Table 3, the calcium content in Samples 1, 5, and 7 was lower than that in the Control 1 (glutaraldehyde-crosslinked porcine pericardium) 30 days after subcutaneous implantation in rats. This result demonstrates that the double-bond post-crosslinking method for preparing bioprosthetic valve materials can improve the anti-calcification properties of bioprosthetic valves.

[0164] Alizarin red staining experiment:

[0165] Samples 1, 5, 7 and control group 1 were implanted subcutaneously in rats for 30 days before being removed and fixed with paraformaldehyde tissue fixative. After fixation, the samples were removed, trimmed flat with a scalpel and transferred to a dehydration box. The material samples were dehydrated in a gradient manner using 50%, 75%, 85%, 95% (v / v) and anhydrous ethanol. After dehydration, the material samples were transferred to an embedding machine and embedded with melted paraffin, then transferred to a -20°C refrigerator for cooling and trimming. 3-5 μm thick sections were cut from the trimmed wax blocks on a microtome, transferred from the slide spreader to a glass slide, and dewaxed and rehydrated. The sections were stained with alizarin red stain for 3 minutes, washed with water, dried, and then permeabilized with xylene for 5 minutes. The sections were sealed with neutral gum and the staining results were imaged on a pathology section scanner.

[0166] The control group 1 (glutaraldehyde cross-linked pig pericardium), sample 1, sample 5, and sample 7 were implanted subcutaneously in rats for 30 days by alizarin red staining to directly observe the degree of calcification of each group of samples. The images of the alizarin red staining results of the sample slices 30 days after implantation in the subcutaneous tissue of rats are shown in Figures 3 to 6, wherein the darker the color of the sample after alizarin red staining, the higher the degree of calcification. Compared with the alizarin red staining results of the slices of the control group 1 (glutaraldehyde cross-linked pig pericardium) (Figure 3), the alizarin red staining images of the slices of sample 1 (Figure 4), sample 5 (Figure 5), and sample 7 (Figure 6) are obviously lighter and paler, which directly indicates that the degree of calcification of sample 1, sample 5, and sample 7 is lower than that of the control group 1, that is, sample 1, sample 5, and sample 7 have a stronger anti-calcification effect than that of the control group 1. The alizarin red staining results of the biological valve material implanted subcutaneously in rats for 30 days show that the method of preparing biological valve material by double bond post-crosslinking of the present application can improve the anti-calcification performance of biological valves.

[0167] Example 11

[0168] Freshly collected porcine pericardium was washed with distilled water at 4°C and 100RPM shaking for 2 hours, immersed in 0.30% (w / w) glutaraldehyde solution at room temperature, and immersed at room temperature for 48 hours to perform glutaraldehyde cross-linking treatment on the biological valve to obtain glutaraldehyde cross-linked porcine pericardium.

[0169] The glutaraldehyde-crosslinked porcine pericardium was washed with deionized water and immersed in a 4% (v / v) ethanol aqueous solution of glycidyl methacrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution was 20% (v / v) ethanol aqueous solution.

[0170] After the double bond modification is completed, ammonium persulfate and sodium bisulfite are added to initiate the polymerization reaction of the double bonds on the double-bonded glutaraldehyde cross-linked pig pericardium, wherein the ammonium persulfate concentration is 20mM and the sodium bisulfite concentration is 5mM; after adding the initiator and reacting at 37°C for 8 hours, the double-bond cross-linked pig pericardium is obtained.

[0171] The pig pericardium material cross-linked after double bond copolymerization was soaked in a 70% ethanol-water solution for 20 minutes, then soaked in a desiccation solution (80% glycerol, 2% water, 18% ethanol) at room temperature for 1.5 hours. Excess glycerol was removed from the surface of the pig pericardium material, and the material was sterilized with ethylene oxide. This was designated as Sample 31.

[0172] Example 12

[0173] Fresh porcine pericardium was placed in a PS solution containing 0.5% sodium deoxycholate (surfactant) by mass, shaken at room temperature for 4 hours, and then washed three times with a 0.9% sodium chloride aqueous solution (ie, normal saline).

[0174] The cleaned porcine pericardium was washed with distilled water at 4°C and 100RPM for 2 hours, and then immersed in 0.30% (w / w) glutaraldehyde solution at room temperature. The bioprosthetic valve was treated with glutaraldehyde cross-linking treatment at room temperature for 48 hours to obtain glutaraldehyde cross-linked porcine pericardium.

[0175] The glutaraldehyde-crosslinked porcine pericardium was further washed with deionized water and immersed in a 5% (v / v) propanol aqueous solution of glycidyl methacrylate at room temperature for double bond modification of the glutaraldehyde-crosslinked porcine pericardium. The reaction time was 72 hours, and the solvent of the double bond modification solution used was a 20% (v / v) propanol aqueous solution.

[0176] After the double bond modification is completed, the double-bond glutaraldehyde cross-linked pig pericardium is washed with deionized water; then the double-bond glutaraldehyde cross-linked pig pericardium is immersed in a mixture of 20mM potassium persulfate and 10mM sodium bisulfite to further induce the polymerization reaction of the double bonds on the double-bond glutaraldehyde cross-linked pig pericardium. After reacting at 37°C for 8 hours, the double-bond post-cross-linked pig pericardium is obtained, which is recorded as sample 34.

[0177] The above-described embodiments merely represent several implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent application. It should be noted that a person of ordinary skill in the art may make various modifications and improvements without departing from the spirit of the present application, and these modifications and improvements fall within the scope of protection of the present application. Therefore, the scope of protection of the present patent application shall be determined by the appended claims.

Claims

1. A method for preparing a biological valve material by double bond polymerization after aldehyde cross-linking, characterized in that: include: Step S110: contacting the biomaterial with an aldehyde cross-linking agent solution for cross-linking; Step S120: Soaking the biomaterial treated in step S110 in a solution containing a first functional monomer to chemically connect the first carbon-carbon double bond; the first functional monomer has a first carbon-carbon double bond and an ethylene oxide group; In step S200 , a carbon-carbon double bond is polymerized under the action of an initiator to obtain a biological valve material.

2. The method according to claim 1, characterized in that The aldehyde cross-linking agent is glutaraldehyde or formaldehyde.

3. The method according to claim 1, characterized in that The biological material is animal tissue, and the animal tissue is selected from one or more of pericardium, valve, intestinal membrane, meninges, lung membrane, blood vessel, skin or ligament.

4. The method according to claim 3, characterized in that The animal tissue is fresh animal tissue or biological tissue that has been decellularized.

5. The method according to claim 1, characterized in that In step S200 : an initiator is added to the system treated in the previous step; or the biological material treated in the previous step is taken out and immersed in a solution containing the initiator directly or after washing.

6. The method according to claim 1, characterized in that The initiator is a single initiator or a mixed initiator.

7. The method according to claim 6, characterized in that The mixed initiator is: A mixture of ammonium persulfate and sodium bisulfite, or a mixture of ammonium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium sulfite, or a mixture of potassium persulfate and sodium sulfite, or a mixture of sodium persulfate and sodium bisulfite, or a mixture of potassium persulfate and sodium bisulfite, or potassium persulfate and tetramethylethylenediamine, or ammonium persulfate and tetramethylethylenediamine, or sodium persulfate and tetramethylethylenediamine; the concentration of each component in the mixture is 1 to 100 mM.

8. The method according to claim 7, characterized in that The single initiator is any component in each mixed initiator.

9. The method according to claim 1, characterized in that In step S200, the double bond polymerization time is 3 to 24 hours.

10. The method according to claim 1, characterized in that The first functional monomer is selected from at least one of allyl glycidyl ether, glycidyl methacrylate and glycidyl acrylate.

11. The method according to claim 1, wherein In step S110: The w / w concentration of the aldehyde cross-linking agent solution is 0.1% to 5%; and the cross-linking time is 0.5h to 120h.

12. The method according to claim 1, characterized in that In step S120: The w / w concentration of the first functional monomer in the solution containing the first functional monomer is 1% to 10%; and the reaction time is 2 to 120 hours.

13. The method according to claim 1, wherein The solution containing the first functional monomer only contains the first functional monomer and a solvent that does not participate in the chemical reaction.

14. The method according to claim 1, wherein The solvent in the solution containing the first functional monomer is one or more of an aqueous solution of any one of methanol, ethanol, ethylene glycol, propanol, 1,2-propylene glycol, 1,3-propylene glycol, isopropanol, butanol, isobutanol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol and glycerol, water, physiological saline, and pH neutral buffer.

15. A biological valve material, characterized in that: The invention is prepared by the method according to any one of claims 1 to 14.

16. A biological valve material, characterized in that: include: Step S110: contacting the biomaterial with an aldehyde cross-linking agent solution for cross-linking; Step S120: Soaking the biomaterial treated in step S110 in a solution containing a first functional monomer to chemically connect the first carbon-carbon double bond; the first functional monomer has a first carbon-carbon double bond and an ethylene oxide group; In step S200 , a carbon-carbon double bond is polymerized under the action of an initiator to obtain a biological valve material.

17. A bioprosthetic valve comprising a stent and a valve leaflet, characterized in that: The leaflet is the biological valve material according to claim 15 or 16.

18. The bioprosthesis according to claim 17, wherein: The biological valve is a heart valve.

19. An interventional system comprising a heart valve and a catheter assembly, wherein the heart valve is folded and then delivered by the catheter assembly, wherein: The heart valve comprises a stent and leaflets, and the leaflets are made of the biological valve material according to claim 15 or 16.

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