CN115920131A

CN115920131A - Biological valve material with anticoagulation and calcification resistance, medical instrument, crosslinking method and application thereof

Info

Publication number: CN115920131A
Application number: CN202111365210.7A
Authority: CN
Inventors: 王云兵; 郑城; 雷洋
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2021-11-17
Filing date: 2021-11-17
Publication date: 2023-04-07

Abstract

The invention discloses a biological valve material with anticoagulation and calcification, a medical device, a cross-linking method and application thereof. The invention can effectively improve the stability, cell compatibility, anticoagulation performance and calcification resistance of biological materials such as biological heart valves and the like, and potentially prolong the service life of the biological heart valves and the like.

Description

Biological valve material with anticoagulation and calcification resistance, medical instrument, crosslinking method and application thereof

Technical Field

The invention relates to a biological valve material with anticoagulation and calcification resistance, a medical instrument, a cross-linking method and application thereof, in particular to a biological valve material with anticoagulation and calcification resistance, a cross-linking method and application thereof, and a medical instrument containing the biological valve material, belonging to the technical field of biomedical materials and medical instruments.

Background

Heart valve disease is a common valve failure disease. Anatomically manifested as narrowing of the blood passageway or incomplete valve closure. Treatment of heart valve disease includes open chest valve replacement surgery and percutaneous heart valve replacement surgery. The thoracotomy has the defects of large wound, high risk, slow recovery and need of extracorporeal circulation support, and is unacceptable for many patients. Percutaneous heart valve replacement surgery is a main trend of valve surgery in the future because of small trauma and low risk to patients.

The biological heart valve is a biomedical material and a device thereof for replacing a diseased heart valve of a human body. The biological heart valve leaflet is generally prepared by cross-linking a porcine pericardium, a bovine pericardium and the like through glutaraldehyde. The glutaraldehyde crosslinking treatment has the characteristics of simple operation, low cost and high collagen crosslinking degree. However, glutaraldehyde cross-linked bioprosthetic heart valves have certain problems of toxicity, material degradation, thrombosis, and calcification due to the presence of residual aldehyde groups on the valve. In the case of damaged valves in young patients, the metabolic capacity of the patient population is great, so that the biological heart valves are more susceptible to calcification and have an effective service life of about 10 years. Therefore, although collagen in the material can be stabilized to a certain extent by the glutaraldehyde crosslinking method, certain cytotoxicity and calcification are caused by the existence of residual aldehyde groups, and the anticoagulation performance of the glutaraldehyde crosslinking method needs to be improved, so that the glutaraldehyde crosslinking biomaterial has certain technical defects.

The invention patent with publication number CN109833519A discloses a method for preparing a biological valve prosthesis, which can prepare a dry biological valve by compounding hydrogel and biological tissue, wherein the preparation method comprises the steps of soaking the biological tissue in a hydrophilic polymer/hydrophilic monomer solution, adding a cross-linking agent, carrying out cross-linking reaction in the presence of a catalyst/initiator to form a three-dimensional network structure, obtaining a biological tissue compounded with the hydrogel, and then carrying out post-crosslinking and dehydration on protein fibers in the biological tissue to obtain the dry biological valve.

In addition, the invention patent with the publication number of CN111481743A discloses an anticoagulant anti-calcification biomaterial and a preparation method thereof, which introduces active groups capable of performing free radical polymerization on biological tissues to perform free radical copolymerization with zwitterionic monomers, so that collagen in the biological tissues is crosslinked at multiple sites through polymers, and full crosslinking in collagen fibers and among fibers is realized, thereby improving the stability of the biological tissues and prolonging the service life of the biological tissues. Meanwhile, zwitterions are introduced to the surface of the biological tissue, so that the anticoagulation performance can be improved, the in-situ endothelialization of the biological valve can be promoted, and the deposition of calcium element is further prevented.

Disclosure of Invention

The invention aims to provide a cross-linking method of a biological valve material with both anticoagulation and calcification, which solves a series of technical defects existing in the existing cross-linking method by adopting isocyano ethyl methacrylate, and can effectively improve the stability, cell compatibility, anticoagulation performance and calcification resistance of biological materials such as biological heart valves and the like by using a polyethylene glycol diacrylate solution, thereby potentially prolonging the service life of the biological heart valves and the like.

The invention is realized by the following technical scheme: a method for cross-linking a biological valve material with both anticoagulation and calcification, comprising the following steps:

s1, soaking the cleaned biological material in an isocyano ethyl methacrylate solution to obtain an isocyano ethyl methacrylate in-situ modified biological valve;

s2, soaking and washing the biological valve obtained in the step S1, and then soaking the biological valve in a polyethylene glycol diacrylate solution to enable the polyethylene glycol diacrylate solution to react with methacrylate groups on the biological valve in the presence of a free radical polymerization initiator to obtain the biological valve material with anticoagulation and calcification resistance.

In the step S1, the in-situ modification of the biological material with the isocyano ethyl methacrylate means that the biological material is soaked in an ethanol solution of the isocyano ethyl methacrylate to enable an amino group in the biological valve material to react with an isocyanate group in the isocyano ethyl methacrylate to realize chemical bonding, so that the methacrylate group capable of free radical polymerization is in-situ modified on the biological valve material.

In the step S2, the reaction in the presence of the radical polymerization initiator means that the radical polymerization initiator initiates a copolymerization reaction between methacrylate groups on the biological valve treated by the isocyanoethyl methacrylate and the polyethylene glycol diacrylate to form a stable carbon-carbon single bond between the collagen of the biological valve material, thereby improving the cross-linking stability and anticoagulation performance of the biological valve material.

In step S1, the biological material is at least one selected from pericardium, valve, intestinal membrane, meninges, lung membrane, blood vessel, skin, and ligament.

In the step S1, when the biomaterial is washed, the pericardial tissue may be washed with distilled water under the oscillation condition by using soft friction and fluid pressure to remove the adhered non-pericardial and non-collagen tissues. Further, effective decellularization of pericardial tissue can be achieved by osmotic shock, with washing continuing until there is no visible adherent non-pericardial or non-collagenous tissue, such as: the washing was carried out with distilled water for 2 hours at 4 ℃ under 100RPM rotational speed shaking.

In the step S1, the volume percentage concentration of the isocyano ethyl methacrylate solution is 0.1-20%.

In the step S1, the solvent in the isocyanoethyl methacrylate solution is at least one selected from ethanol, a mixed solution of tween and water, a mixed solution of ethanol and water, and isopropanol or a mixed solution of isopropanol and water.

Further, an ethanol solution of isocyanoethyl methacrylate at a concentration of 0.1 to 10% may be used, the treatment time is 0.5 to 24 hours, and the temperature is 0 ℃.

In the step S2, the volume percentage concentration of the polyethylene glycol diacrylate solution is 0.1-10%.

In the step S2, the radical polymerization initiator is at least one selected from ammonium persulfate-sodium bisulfite, potassium persulfate-sodium bisulfite, ammonium persulfate-potassium bisulfite, potassium persulfate-potassium bisulfite, ammonium persulfate-tetramethylethylenediamine, and potassium persulfate-tetramethylethylenediamine.

In the step S2, the concentration of the radical polymerization initiator is 1 to 100mM.

In the step 2, the biological valve can be soaked and washed by distilled water and ethanol, and the biological valve material obtained by the reaction is also washed by distilled water and ethanol to remove other non-covalently bonded impurities adhered to the surface of the biological valve material.

The invention also provides a biological valve material prepared by the cross-linking method, and application of the biological valve material, which is used for preparing a percutaneous intervention biological heart valve material or a biological valve material for an open-chest valve.

The invention also provides a medical appliance containing the biological valve material, which comprises an interventional biological valve, an artificial blood vessel, an artificial skin, an artificial cardiac muscle patch and a dura mater.

Compared with the prior art, the invention has the following advantages and beneficial effects:

the invention provides a biological valve material with anticoagulation and calcification, which is characterized in that a strong electrophilic isocyanate group in isocyano ethyl methacrylate reacts with a nucleophilic functional group such as an amino group on the biological valve material to form a stable chemical bond (urea bond) and introduce a methacrylate group capable of free radical polymerization, the valve material treated by the isocyano ethyl methacrylate is further soaked in a hydrophilic monomer polyethylene glycol diacrylate solution, and finally, the crosslinking of the biological valve material is realized by adding a free radical polymerization initiator to initiate the free radical polymerization, so that the stability, the cell compatibility and the calcification resistance of the biological valve can be improved, the anticoagulation performance of the biological valve can be improved, and the problem of short service life of the biological valve caused by cytotoxicity, material degradation and calcification is solved.

Drawings

FIG. 1 is a specific flow chart of the preparation method of the biological valve material with anticoagulation and calcification resistance.

Fig. 2 is a schematic diagram of the principle of preparing a biological valve material with anticoagulation and calcification according to the present invention.

FIG. 3 is a result of a platelet adhesion experiment of a crosslinked biological valve material.

FIG. 4 shows the enzyme degradation weight loss ratio of the crosslinked biological valve material.

Fig. 5 is a graph of cytotoxicity of the crosslinked bioprosthetic valve material.

FIG. 6 shows the anticalcification performance of crosslinked biological valve material (alizarin red stain).

FIG. 7 is a graph of the anticalcification performance (elemental calcium content) of the crosslinked biological valve material.

FIG. 8 is the anticoagulant property of the crosslinked biological valve material (scanning electron micrograph of platelet adhesion).

FIG. 9 is the anticoagulant property of the crosslinked biological valve material (whole blood adhesion scanning electron microscopy).

Detailed Description

The objects, technical solutions and advantageous effects of the present invention will be described in further detail below.

It is to be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention claimed, and unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

In the existing glutaraldehyde crosslinking method, aldehyde groups are remained on the valve, so that materials such as a biological valve crosslinked by the glutaraldehyde crosslinking method have the problems of certain toxicity, material degradation, thrombus and calcification. Therefore, there is a need to research and develop a new crosslinking method for a biological valve material.

The biological valve material with both anticoagulation and calcification can be prepared by adopting an isocyano ethyl methacrylate crosslinking method and introducing polyethylene glycol diacrylate, and the principle of the biological valve material is shown in figure 1. The isocyano ethyl methacrylate is a cross-linking agent with high reactivity with a biological valve material, and the structure of the cross-linking agent has a strong electrophilic isocyanate group and a free radical polymerizable methacrylate group. Firstly, isocyanate groups and nucleophilic groups on a biological valve material can react to form stable urea bonds, and methacrylate groups capable of free radical polymerization are covalently introduced to the biological valve material; secondly, the methacrylate group capable of free radical polymerization can form a relatively stable carbon-carbon single bond under the action of a free radical polymerization initiator so as to realize stable crosslinking of the biological valve material; moreover, compared with reversible Schiff base bonds of the glutaraldehyde crosslinking agent in crosslinking, the reaction activity of isocyanate groups and amino groups in the biological valve material is higher, the formed urea bonds have higher stability, and the collagen components of the biological valve material can be better stabilized; finally, compared with a glutaraldehyde crosslinking agent, the isocyano ethyl methacrylate crosslinking agent has no toxic residual aldehyde groups, and can potentially avoid the problems of cytotoxicity, calcification and the like caused by the residual aldehyde groups. Therefore, the isocyano ethyl methacrylate can be used for crosslinking of the biological valve material so as to improve the stability of the material and the biocompatibility of the calcification-resistant performance. Research shows that isocyanate groups of isocyanate compounds can react with amino groups in collagen biomacromolecules to realize crosslinking of biomaterials.

Aiming at the problems of thrombus caused by blood coagulation on the biological valve and the like, the strategy of performing anticoagulation drug intervention to prevent blood coagulation events after operation is adopted, and the anticoagulation modification treatment on the biological valve is also favorable for reducing the risk of thrombus formation. Protein adhesion occurs firstly when the biological material is contacted with blood, and the adhesion of platelets and the activation of a blood coagulation system are further mediated through the formation of a protein layer on the surface of the material, so that the formation of thrombus is promoted. Typically, anticoagulant modification of biological materials is primarily by improving the hydrophilicity of the material. By introducing hydrophilic molecules on the material, the material is made more hydrophilic. The hydrophilic surface is beneficial to forming a hydration layer when contacting blood so as to be lower than protein adhesion, thereby reducing the reaction of biological materials and blood, such as blood cell adhesion and the like, and further reducing the aim of anticoagulation. Polyethylene glycol is a molecule with good hydrophilicity and good blood compatibility, and is commonly used for modifying biological materials to reduce the incidence of coagulation events by improving the hydrophilicity of the materials. The polyethylene glycol diacrylate also comprises acrylate groups capable of free radical polymerization besides a hydrophilic polyethylene glycol structure, so that the polyethylene glycol diacrylate is favorable for carrying out free radical copolymerization reaction with biological materials with free radical polymerization groups to realize pegylation modification of the materials so as to improve the anticoagulation performance of the materials and reduce the incidence rate of thrombosis events.

Therefore, by adopting the method of the biological valve material modified by the isocyano ethyl methacrylate, the isocyanate group in the isocyano ethyl methacrylate is reacted with the amino group in the biological valve material to connect the free radical polymerizable methacrylate group onto the biological valve material through a stable chemical bond; and then, carrying out free radical copolymerization on the isocyano ethyl acrylate-modified biological valve material and polyethylene glycol diacrylate under the action of a free radical polymerization initiator to realize pegylation modification of the biological valve material so as to improve the anticoagulation performance of the biological valve material. On one hand, the cross-linking agent without aldehyde residue is used, calcification of biological valve materials related to aldehyde is avoided, on the other hand, because isocyanate groups and amino groups react to form stable urea bonds, the stability of the biological heart valve is improved by utilizing a isocyano ethyl methacrylate cross-linking method, and moreover, polyethylene glycol diacrylate macromolecules are introduced while cross-linking, so that the anticoagulation performance of the biological heart valve is improved.

In the prior art, patent publication No. CN109833519a discloses that isocyanate compounds can be used as cross-linking agents for preparing cross-linked bioprosthetic valves, and the reaction mechanism is to react the isocyanate compounds with hydrophilic polymers, thereby cross-linking protein fibers in biological tissues. However, in the method of the present invention, isocyanate in isocyano ethyl methacrylate as a cross-linking agent reacts with nucleophilic groups on the biological valve material to form relatively stable urea bonds, and methacrylate groups capable of free radical polymerization are introduced, and the method is characterized in that the biological valve material is soaked in an isocyanate solution and then methacrylate groups are introduced, so that the reaction mechanism of the two is different from the invention motivation.

In the prior art, the invention patent with publication number CN111481743A discloses that active group methacrylate group is introduced into biological tissue, but the function of the invention is to generate radical copolymerization with zwitterionic monomer, so that collagen in the biological tissue is crosslinked through multiple points of polymer, and full crosslinking in collagen fiber and among fibers is realized, wherein zwitter ions are connected with biological valve material through amide bond. However, in the method of the present invention, due to the introduction of the isocyanate group, the active group methacrylate group can be connected with the biological valve through a stable urea bond, and finally, through the introduction of the polyethylene glycol diacrylate, the cross-linked structure of the polyethylene glycol diacrylate polymer network is obtained. Therefore, the crosslinking structures to be prepared are not the same.

In conclusion, the biological valve material is prepared by copolymerizing the isocyano ethyl methacrylate modified biological valve material and the polyethylene glycol diacrylate, so that the stability, the cell compatibility and the calcification-resisting performance of the biological valve can be improved, and the anticoagulation performance can also be improved, so that the problem that the service life of the biological valve is short due to cytotoxicity, material degradation and calcification is potentially solved, and relevant reports are not found in the prior art.

The following examples are provided to illustrate specific embodiments of the present invention, and reference is made to the process flow shown in fig. 2, but the scope of the present invention is not limited to the following examples.

In all of the following embodiments, the fresh porcine/bovine pericardium is from a local slaughterhouse. Isocyanoethyl methacrylate (2-Isocyanoethyl methacrylate, ICM), glutaraldehyde (GA), ammonium Persulfate (APS), and Sodium bisulfite (SHS) were obtained from Sigma-Aldrich. Polyethylene glycol diacrylate (PEGDA) is from chenopodium chemie company.

Example 1:

freshly collected pig hearts were washed with distilled water for 2 hours at 4 ℃ under 100RPM shaking. Then soaked in 10% isocyano ethyl methacrylate ethanol solution and treated at 4 ℃ for 24 hours. Wash 3 times with distilled water/ethanol for 10 minutes each time. The pericardium was then soaked in a 1% by volume aqueous solution of polyethylene glycol diacrylate for 4 hours. The ammonium persulfate/sodium bisulfite aqueous solution was treated at 37 degrees Celsius for 24 hours at a concentration of 40mM. And finally, soaking the pericardium soaked by the ammonium persulfate/sodium bisulfite aqueous solution, and washing the pericardium with distilled water/ethanol to obtain the pericardium material which is recorded as PICM-PEGDA1-PP.

Example 2:

freshly collected pig hearts were washed with distilled water for 2 hours at 4 ℃ under 100RPM shaking. Then soaked in 10% isocyano ethyl methacrylate ethanol solution and treated at 4 ℃ for 24 hours. Wash 3 times with distilled water/ethanol for 10 minutes each time. The pericardium was then soaked in a 3% by volume aqueous solution of polyethylene glycol diacrylate for 4 hours. The ammonium persulfate/sodium bisulfite aqueous solution was treated at 37 degrees Celsius for 24 hours at a concentration of 40mM. And finally, soaking the pericardium soaked by the ammonium persulfate/sodium bisulfite aqueous solution, and washing the pericardium with distilled water/ethanol to obtain the pericardium material which is recorded as PICM-PEGDA3-PP.

Example 3:

freshly collected pig hearts were washed with distilled water for 2 hours at 4 ℃ under 100RPM shaking. Then soaked in 10% isocyano ethyl methacrylate ethanol solution and treated at 4 ℃ for 24 hours. Wash 3 times with distilled water/ethanol for 10 minutes each time. The pericardium was then soaked in a 5% by volume aqueous solution of polyethylene glycol diacrylate for 4 hours. The ammonium persulfate/sodium bisulfite aqueous solution was treated at 37 degrees Celsius for 24 hours at a concentration of 40mM. And finally, soaking the pericardium soaked by the ammonium persulfate/sodium bisulfite aqueous solution, and washing the pericardium with distilled water/ethanol to obtain the pericardium material which is recorded as PICM-PEGDA5-PP.

Example 4:

freshly harvested pig hearts were washed with distilled water at 4 ℃ for 2 hours under 100RPM speed shaking. Then soaked in 10% isocyano ethyl methacrylate ethanol solution and treated at 4 ℃ for 24 hours. Wash 3 times with distilled water/ethanol for 10 minutes each time. The pericardium was then soaked in 7% by volume aqueous polyethylene glycol diacrylate for 4 hours. The ammonium persulfate/sodium bisulfite aqueous solution was treated at 37 degrees Celsius for 24 hours at a concentration of 40mM. And finally, soaking the pericardium soaked by the ammonium persulfate/sodium bisulfite aqueous solution, and washing the pericardium with distilled water/ethanol to obtain the pericardium material which is recorded as PICM-PEGDA7-PP.

Example 5:

the material obtained in example 1 is used for preparing a percutaneous interventional biological heart valve material, and in addition to the required biological valve material, other medical materials required by percutaneous interventional surgery also can be contained.

Example 6:

the material obtained in example 2 is used for preparing a biological valve material for replacement of an open chest valve, and in addition to the required biological valve material, the material also comprises other medical materials required by the open chest valve replacement surgery.

Example 7:

a medical device comprising the material obtained in example 3, which may be an interventional biovalve, a vascular prosthesis, a skin prosthesis, a myocardial patch prosthesis or dura mater, is prepared to comprise other medically desirable materials in addition to the desired biovalve material.

During the treatment, the following two control groups were set:

glutaraldehyde (GA) treatment group: i.e. the pericardium is soaked in 0.625% glutaraldehyde for 24 hours.

Isocyanoethyl methacrylate (ICM) direct cross-linked group: namely, the freshly collected pig hearts are washed with distilled water for 2 hours under the condition of 4 ℃ and 100RPM rotational speed oscillation. Then soaking the mixture in 10 volume percent ethanol solution of isocyano ethyl methacrylate, and treating the mixture for 24 hours at 4 ℃. Wash 3 times with distilled water/ethanol for 10 minutes each time. The pericardium is then soaked in an ammonium persulfate/sodium bisulfite aqueous solution with a concentration of 40mM, and treated at 37 ℃ for 24 hours. And finally, washing the pericardium soaked by the ammonium persulfate/sodium bisulfite water solution with distilled water/ethanol, and recording the obtained biological valve material as ICM.

The materials obtained in examples 1 to 4 were subjected to a quantitative analysis of platelet adhesion with the materials of the Glutaraldehyde (GA) -treated group and the isocyano ethyl methacrylate (ICM) -directly crosslinked group. And cutting the materials obtained by different treatment modes into sheets with consistent sizes to perform a platelet adhesion test. The amount of platelet adhesion was characterized by measuring the Relative amount of lactate dehydrogenase (Relative LDH Content) released from the surface of each group of samples according to a reported method (Biomacromolecules 2021,22,823-836). The difference in the amount of adhered platelets on different materials was characterized by detecting lactate dehydrogenase released from platelets adhering to the surface of the material by lactate dehydrogenase kit (LDH kit). The results are shown in fig. 3, where the relative lactate dehydrogenase LDH content of the peg diacrylate co-crosslinked pericardium material was lower than that of the glutaraldehyde crosslinked pericardium, where a decrease in lactate dehydrogenase release was indicated by an increase in peg diacrylate concentration used during crosslinking, indicating a decrease in platelet adhesion. Reduced platelet adhesion is beneficial in reducing the probability of clotting events. The result shows that the adhesion amount is relatively low when the concentration of the polyethylene glycol diacrylate is 5%, and the pericardium treatment with the concentration of the polyethylene glycol diacrylate being 5% and covalently crosslinked is further subjected to enzyme degradation stability characterization, cytotoxicity characterization, anti-calcification performance, platelet adhesion and whole blood adhesion characterization.

The component stability of the crosslinked bioprosthetic valve material was examined by collagenase degradation experiments. Degradation experiments were performed on the crosslinked samples, and the results are shown in fig. 4, where the weight loss rates of the GA group, the ICM group, and the ICM-PEGDA5 group were 9.3%,2.1%, and 1.1%, respectively. The pericardium ICM-PEGDA5 which is subjected to covalent cross-linking by polyethylene glycol diacrylate has the lowest collagenase degradation Weight Loss Ratio (Weight Loss Ratio), which shows that the component stability is higher.

Cytotoxicity experiments showed that, as shown in fig. 5, the GA group showed significant cytotoxicity as indicated by lower cell viability, whereas both the ICM-PEGDA5 and ICM groups showed no significant cytotoxicity and lower cell viability.

Samples of GA group, ICM group and ICM-PEGDA5 group were implanted into rats subcutaneously for 60 days and then taken out for alizarin red staining and calcium ion content analysis to characterize the anti-calcification performance. Alizarin red stains calcified deposits dark red, with darker colors indicating more calcifications and lighter colors indicating less calcifications. Alizarin red staining results are shown in FIG. 6, and ICM-PEGDA5 group is light in color and has no obvious deposition, which shows that ICM-PEGDA5 has calcification-resisting performance relative to GA group. Calcium analysis the ICM-PEGDA5 group showed the lowest calcium content, as shown in FIG. 7. The calcification-resistant performance analysis shows that the pericardium ICM-PEGDA5 which is covalently crosslinked by the polyethylene glycol diacrylate has better calcification-resistant performance.

Platelet adhesion and whole blood adhesion tests were performed on the crosslinked samples to characterize anticoagulant properties. As shown in fig. 8 and 9, the pericardium ICM-PEGDA5 covalently cross-linked with polyethylene glycol diacrylate showed less adhesion of platelets and whole blood, and showed resistance to adhesion of blood cells, which is advantageous in reducing the occurrence of inhibition of coagulation reaction and thus the probability of thrombosis, compared to the GA and ICM groups. Platelet adhesion and whole blood adhesion tests prove that the pericardium ICM-PEGDA5 subjected to covalent cross-linking by polyethylene glycol diacrylate has the anticoagulant property.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications and equivalent variations of the above embodiments according to the technical spirit of the present invention are included in the scope of the present invention.

Claims

1. A cross-linking method of a biological valve material with anticoagulation and calcification resistance is characterized in that: the method comprises the following steps:

s2, soaking and washing the biological valve obtained in the step S1, and then soaking the biological valve in a polyethylene glycol diacrylate solution to enable the polyethylene glycol diacrylate solution to react with methacrylate groups on the biological valve in the presence of a free radical polymerization initiator, so as to obtain the biological valve material with both anticoagulation and calcification resistance.

2. The crosslinking method according to claim 1, wherein: in step S1, the biological material is at least one selected from pericardium, valve, intestinal membrane, meninges, lung membrane, blood vessel, skin, and ligament.

3. The crosslinking method according to claim 1, wherein: in the step S1, the volume percentage concentration of the isocyano ethyl methacrylate solution is 0.1-20%.

4. The crosslinking method according to claim 1, wherein: in the step S1, the solvent in the isocyanoethyl methacrylate solution is at least one selected from ethanol, a mixed solution of tween and water, a mixed solution of ethanol and water, and isopropanol or a mixed solution of isopropanol and water.

5. The crosslinking method according to claim 1, wherein: in the step S2, the volume percentage concentration of the polyethylene glycol diacrylate solution is 0.1-10%.

6. The crosslinking method according to claim 1, wherein: in the step S2, the radical polymerization initiator is at least one selected from ammonium persulfate-sodium bisulfite, potassium persulfate-sodium bisulfite, ammonium persulfate-potassium bisulfite, potassium persulfate-potassium bisulfite, ammonium persulfate-tetramethylethylenediamine, and potassium persulfate-tetramethylethylenediamine.

7. The crosslinking method according to claim 1, wherein: in the step S2, the concentration of the radical polymerization initiator is 1 to 100mM.

8. A bioprosthetic valve material prepared by the crosslinking method of any one of claims 1 to 7.

9. Use of the bioprosthetic valve material of claim 8, wherein: is used for preparing percutaneous intervention biological heart valve material or biological valve material for open valve placement.

10. A medical device comprising the biological valve material of claim 8, wherein: including biological valve, artificial blood vessel, artificial skin, artificial cardiac muscle patch and dura mater.