Plasma substitute surface engineering bioprosthetic valve material and preparation method thereof

Abstract

The invention discloses a plasma substitute surface engineering bioprosthetic valve material and a preparation method thereof, the preparation method comprises the following steps: S1, obtaining a bioprosthetic valve material, and pretreating the bioprosthetic valve material; s2, activating the pretreated bioprosthetic valve material by adopting an amide condensing agent to obtain a carboxyl activated bioprosthetic valve material; s3, grafting the bioprosthetic valve material after the carboxyl is activated by adopting 2-aminoethyl methacrylate hydrochloride to obtain a bioprosthetic valve material grafted with a methacrylate group; and S4, under the action of an initiator, carrying out polymerization reaction on methacrylated hydroxyethyl starch and the bioprosthetic valve material grafted with the methacrylate group, and cleaning to obtain the plasma substitute surface-engineered bioprosthetic valve material. According to the invention, the defects of toxicity, calcification, immunoreaction and the like caused by glutaraldehyde crosslinking of the existing bioprosthetic valve can be overcome, and the defects of thrombus and immunoreaction caused by heterogeneous collagen matrix of the bioprosthetic valve material can be overcome.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials and medical device technology, and in particular to a plasma substitute surface engineered bio-valve material and its preparation method. Background Technology

[0002] Valvular heart disease is a common cardiovascular disease, primarily characterized by stenosis and regurgitation, with high morbidity and mortality. With the accelerating aging of the population, the demand for artificial heart valve replacement surgery is increasing year by year. Compared to mechanical valves, bioprosthetic valves exhibit superior hemodynamic performance, and patients do not require long-term anticoagulation therapy after implantation, making them more popular among patients, especially those unsuitable for long-term anticoagulation. In recent years, with the widespread clinical application and continuous development of transcatheter aortic valve replacement, the complexity and risks of bioprosthetic valve implantation surgery have significantly decreased, providing a safer and more reliable treatment option for elderly patients. It is foreseeable that with the intensifying trend of population aging, the clinical application of bioprosthetic valves will become increasingly widespread.

[0003] Clinically used implantable bioprosthetic valves are primarily made from xenogeneic tissue materials such as porcine or bovine pericardium treated with glutaraldehyde. Glutaraldehyde cross-linking significantly improves the stability and mechanical properties of bioprosthetic valve materials. However, on the one hand, the toxic aldehyde residue resulting from glutaraldehyde cross-linking leads to defects in bioprosthetic valves, including toxicity, calcification, and immune responses. On the other hand, the xenogeneic collagen matrix of the bioprosthetic valve material exposes it to thrombosis and immune responses. These defects cause structural degeneration of bioprosthetic valves within 10-15 years after implantation, leading to failure and decay, thus increasing the risk of secondary valve replacement surgery for patients. The structural degeneration of bioprosthetic valves is closely related to the glutaraldehyde cross-linking method and the cytotoxicity, calcification, thrombosis, and immune responses induced by the xenogeneic collagen matrix. Therefore, to reduce the risk of structural degeneration of bioprosthetic valves and avoid premature failure and decay, it is necessary to overcome the two major technical defects of existing bioprosthetic valves: the defects caused by glutaraldehyde cross-linking and the xenogeneic collagen matrix of the bioprosthetic valve material.

[0004] Furthermore, since bioprosthetic valve materials are collagen-based biomaterials, they contain residues such as amino, hydroxyl, and carboxyl groups. The carboxyl group readily ionizes into a carboxylate group, which binds to calcium ions, promoting calcium deposition on the bioprosthetic valve and thus facilitating calcification. This is another risk associated with bioprosthetic valve calcification. However, current glutaraldehyde crosslinking achieves crosslinking of bioprosthetic valve materials by consuming amino groups, but it cannot consume carboxyl groups. Therefore, while overcoming the shortcomings of glutaraldehyde crosslinking and the heterogeneous collagen matrix of bioprosthetic valve materials, it is also urgent to consume the carboxyl groups on bioprosthetic valve materials to reduce the risk of calcification. Summary of the Invention

[0005] To address the aforementioned problems, this invention aims to provide an engineered bio-valve material for the surface of blood plasma substitutes and its preparation method.

[0006] The technical solution of the present invention is as follows: On the one hand, a method for preparing engineered bio-valve material for the surface of blood plasma substitutes is provided, comprising the following steps: S1: Obtain bioprosthetic valve material and pretreat the bioprosthetic valve material; S2: The pretreated bioprosthetic valve material is activated with an amide condensing agent to obtain bioprosthetic valve material with activated carboxyl groups; S3: The activated carboxyl group of the biovalve material is grafted with 2-aminoethyl methacrylate hydrochloride to obtain a biovalve material grafted with methacrylate groups; S4: Under the action of an initiator, methacrylamide hydroxyethyl starch is polymerized with the bio-valve material grafted with methacrylate groups, and then washed to obtain the plasma substitute surface engineered bio-valve material.

[0007] Preferably, in step S1, the bioprosthetic valve material is any one of the following: pericardium, valve, intestinal membrane, meninges, pulmonary membrane, blood vessel, skin, amnion, and ligament.

[0008] Preferably, in step S1, the preprocessing includes the following sub-steps: S11: The obtained bioprosthetic valve material was stored in physiological saline under low temperature and humid conditions at 0-5℃; S12: Wash the bioprosthetic valve material preserved in step S11 with physiological saline on a shaker until there are no visible adhering non-pericardial or non-collagenous tissues and residues.

[0009] Preferably, in step S2, the amide condensing agent is a combination of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) or 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM).

[0010] Preferably, step S2 specifically includes the following sub-steps: S21: Prepare an amide condensing agent solution; S22: Immerse the pretreated bio-valve material in the amide condensing agent solution to bring the amide condensing agent into contact with the bio-valve material to activate the carboxyl groups on the bio-valve material, thereby obtaining the bio-valve material with activated carboxyl groups.

[0011] Preferably, in step S21, the concentration of the amide condensing agent solution is 0.05-0.5M; in step S22, the soaking time is 0.1-1h and the temperature is 4-40℃.

[0012] Preferably, step S3 specifically includes the following sub-steps: adding 2-aminoethyl methacrylate hydrochloride to a solution of the bio-valve material soaked with the activated carboxyl group, and then reacting at 4-50°C for 12-24 hours to obtain the bio-valve material grafted with the methacrylate group. Step S4 specifically includes the following sub-steps: S41: Prepare a solution of methacrylated hydroxyethyl starch; S42: The bio-valve material grafted with methacrylate groups is immersed in the methacryloyl hydroxyethyl starch solution, an initiator is added and the reaction is carried out for 5-24 hours, followed by washing to obtain the plasma substitute surface engineered bio-valve material.

[0013] Preferably, in step S3, the mass concentration of the 2-aminoethyl methacrylate hydrochloride is 1%-5%; and in step S41, the concentration of the methacrylated hydroxyethyl starch solution is 1%-20%.

[0014] Preferably, in step S4, the initiator is a redox initiator, wherein the oxidant of the redox initiator is any one or more of sodium persulfate, ammonium persulfate, and potassium persulfate; the reducing agent of the redox initiator is any one or more of sodium sulfite, sodium bisulfite, and potassium bisulfite; and the concentrations of the oxidant and the reducing agent are both 2-50 mM.

[0015] On the other hand, a blood substitute surface engineered bio-valve material is also provided, which is prepared by the preparation method of the blood substitute surface engineered bio-valve material described in any one of the above-mentioned methods.

[0016] The beneficial effects of this invention are: On the one hand, this invention avoids the use of glutaraldehyde crosslinking agent, which can overcome the defects of existing bioprosthetic valves caused by glutaraldehyde crosslinking, such as toxicity, calcification and immune response; on the other hand, by introducing hydroxyethyl starch (plasma substitute) into the bioprosthetic valve material, this invention can overcome the defects of thrombosis and immune response caused by the xenogeneic collagen matrix of bioprosthetic valve material. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 This is a schematic diagram illustrating the principle of the preparation method of the engineered bio-valve material on the surface of the blood plasma substitute of the present invention; Figure 2 This is a schematic diagram showing the fluorescence staining results of the cytotoxicity experiment of the bioprosthetic valve materials in Comparative Example 1 and Example 1; Figure 3 This is a schematic diagram showing the blood adhesion (antithrombotic) test results of the bioprosthetic valve materials in Comparative Example 1 and Example 1; Figure 4 This is a schematic diagram of the H&E staining results of the rat subcutaneous implantation experiment of the bioprosthetic valve material in Comparative Example 1 and Example 1; Figure 5 This is a schematic diagram showing the results of alizarin red (calcification) staining in rat subcutaneous implantation experiments of bioprosthetic valve materials in Comparative Example 1 and Example 1. Figures 2-5 In the diagram, (a) is a schematic diagram of the results of Comparative Example 1; (b) is a schematic diagram of the results of Example 1. Detailed Implementation

[0019] The present invention will be further described below with reference to the accompanying drawings and embodiments. It should be noted that, unless otherwise specified, the embodiments and technical features described in this application can be combined with each other. It should also be pointed out that, unless otherwise indicated, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. The terms "comprising" or "including" and similar words used in this invention refer to elements or objects preceding the word that encompass the elements or objects listed following the word and their equivalents, without excluding other elements or objects.

[0020] On the one hand, such as Figure 1 As shown, this invention provides a method for preparing engineered bio-valve material for the surface of blood plasma substitutes, comprising the following steps: S1: Obtain bioprosthetic valve material and pretreat the bioprosthetic valve material; S2: The pretreated bioprosthetic valve material is activated with an amide condensing agent to obtain bioprosthetic valve material with activated carboxyl groups; S3: The activated carboxyl group of the biovalve material is grafted with 2-aminoethyl methacrylate hydrochloride to obtain a biovalve material grafted with methacrylate groups; S4: Under the action of an initiator, methacrylamide hydroxyethyl starch is polymerized with the bio-valve material grafted with methacrylate groups, and then washed to obtain the plasma substitute surface engineered bio-valve material.

[0021] In this invention, the carboxyl groups of the bioprosthetic valve material are first activated by an amide condensing agent, followed by an amide condensation reaction with the amino group of 2-aminoethyl methacrylate hydrochloride. This covalently introduces methacrylate onto the bioprosthetic valve material and consumes the carboxyl groups, shielding sites on the bioprosthetic valve that readily bind calcium ions, thus reducing calcium ion deposition and lowering the risk of calcification. Furthermore, an initiator is added to initiate a polymerization reaction between the bioprosthetic valve material and the methacrylate groups on methacryloyl hydroxyethyl starch, simultaneously achieving crosslinking of the bioprosthetic valve material and introducing hydroxyethyl starch onto the bioprosthetic valve surface, constructing a non-glutaraldehyde crosslinked plasma substitute surface-engineered bioprosthetic valve material.

[0022] This invention introduces methacrylate groups by consuming carboxyl groups on bioprosthetic valve materials that readily bind calcium ions. Through polymerization between the bioprosthetic valve material and the methacrylate groups on methacryloyl hydroxyethyl starch, the bioprosthetic valve material is cross-linked, and a plasma substitute surface is constructed. This achieves the effects of eliminating calcification-promoting carboxyl sites on the bioprosthetic valve material, avoiding glutaraldehyde cross-linking defects, and using a plasma substitute coating to shield the heterogeneous collagen matrix of the bioprosthetic valve material. It overcomes the defects of existing glutaraldehyde-crosslinked bioprosthetic valve materials, such as toxicity, thrombosis, calcification, and immune reactions, and reduces the risk of structural degeneration of bioprosthetic valves.

[0023] In one specific embodiment, in step S1, the bioprosthetic valve material is any one of the following: pericardium, valve, intestinal membrane, meninges, pulmonary membrane, blood vessel, skin, amnion, and ligament. It should be noted that the bioprosthetic valve material in this embodiment is only a preferred bioprosthetic valve material for this embodiment; other animal tissue materials suitable as bioprosthetic valves in the prior art can also be applied to this invention.

[0024] In a specific embodiment, step S1, the preprocessing includes the following sub-steps: S11: The obtained bioprosthetic valve material was stored in physiological saline under low temperature and humid conditions at 0-5℃; S12: The bioprosthetic valve material preserved in step S11 is washed with physiological saline on a shaker until there is no visible adhering non-pericardial or non-collagenous tissue or residue; optionally, it is washed on a shaker at a speed of 50-200 rpm for 1-5 hours.

[0025] It should be noted that the purpose of pretreating the bioprosthetic valve material is to remove non-pericardial or non-collagenous tissue adhering to the surface of the bioprosthetic valve material. The pretreating method in the above embodiment is only one preferred method to achieve this purpose. Other pretreating methods in the prior art that can achieve this purpose can also be applied to the present invention.

[0026] In one specific embodiment, in step S2, the amide condensing agent is a combination of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) or 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM).

[0027] It should be noted that the purpose of the amide condensing agent is to activate the carboxyl groups of the bioprosthetic valve material. In addition to the amide condensing agent preferred in the above embodiments, other amide condensing agents in the prior art that can also achieve this purpose are also applicable to the present invention.

[0028] In a specific embodiment, step S2 specifically includes the following sub-steps: S21: Prepare an amide condensing agent solution; optionally, the concentration of the amide condensing agent solution is 0.05-0.5M; S22: The pretreated bioprosthetic valve material is immersed in the amide condensing agent solution. Optionally, the immersion time is 0.1-1 h and the temperature is 4-40 °C, so that the amide condensing agent comes into contact with the bioprosthetic valve material to activate the carboxyl groups on the bioprosthetic valve material, thereby obtaining the bioprosthetic valve material with activated carboxyl groups.

[0029] In a specific embodiment, step S3 specifically includes the following sub-steps: adding 2-aminoethyl methacrylate hydrochloride to a solution of the bioprosthetic valve material soaked with the activated carboxyl group, and then reacting at 4-50°C for 12-24 hours to obtain the bioprosthetic valve material grafted with the methacrylate group; optionally, the mass concentration of the 2-aminoethyl methacrylate hydrochloride is 1%-5% (w / v); optionally, the solvent for soaking the bioprosthetic valve material after activating the carboxyl group is any one or more of deionized water, physiological saline, PBS buffer, MES buffer, ethanol, and isopropanol.

[0030] In a specific embodiment, step S4 specifically includes the following sub-steps: S41: Prepare a methacrylated hydroxyethyl starch solution; optionally, the concentration of the methacrylated hydroxyethyl starch solution is 1%-20% (w / v).

[0031] S42: The bio-valve material grafted with methacrylate groups is immersed in the methacryloyl hydroxyethyl starch solution, an initiator is added and the reaction is carried out for 5-24 hours, followed by washing to obtain the plasma substitute surface engineered bio-valve material.

[0032] It should be noted that, in order to ensure that the methacrylamide hydroxyethyl starch can fully contact and penetrate the bioprosthetic valve material, the bioprosthetic valve material grafted with methacrylate groups is immersed in the methacrylamide hydroxyethyl starch solution and kept immersed for 1-2 hours before the initiator is added to carry out the reaction.

[0033] In a specific embodiment, in step S4, the initiator is a redox initiator, wherein the oxidant of the redox initiator is any one or more of sodium persulfate, ammonium persulfate, and potassium persulfate; the reducing agent of the redox initiator is any one or more of sodium sulfite, sodium bisulfite, and potassium bisulfite; and the concentrations of the oxidant and the reducing agent are both 2-50 mM.

[0034] It should be noted that the purpose of the initiator is to enable the bio-valve material grafted with methacryloyl hydroxyethyl starch to undergo a polymerization reaction under the action of the initiator. In addition to the preferred initiator in the above embodiments, other initiators in the prior art that can achieve this purpose are also applicable to the present invention.

[0035] On the other hand, the present invention also provides a plasma substitute surface engineered bio-valve material, which is prepared by the preparation method of plasma substitute surface engineered bio-valve material described in any one of the above-mentioned methods.

[0036] In one specific embodiment, the blood plasma substitute surface engineered bio-valve material described in this invention is used to fabricate heart valves and venous valves.

[0037] Example 1 A blood plasma substitute surface engineered bio-valve material is prepared through the following steps: Freshly collected porcine pericardium was washed with physiological saline at 4°C and 100 rpm for 1 hour to remove non-collagenous tissues such as fat. The pericardium was then activated at 37°C for 0.5 hours by immersing it in a deionized aqueous solution containing 0.1 M 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS). 4% 2-aminoethyl methacrylate hydrochloride was further added, and the reaction was carried out at 37°C for 12 hours. The pericardium was then transferred to a solution containing 5% methacryloyl hydroxyethyl starch and soaked for 1 hour. Finally, 20 mM potassium persulfate and 10 mM sodium sulfite were added, and the reaction was carried out at 37°C for 6 hours. The pericardium was then washed with physiological saline to obtain the non-glutaraldehyde crosslinked plasma-modified bio-valve material.

[0038] In this embodiment, the methacrylated hydroxyethyl starch is prepared by the following steps: 10g of hydroxyethyl starch is dissolved in 100mL of deionized water, then 10mL of glycidyl methacrylate is added and reacted at room temperature for 48h. After dialyzing for 5d, the methacrylated hydroxyethyl starch is obtained by freeze drying.

[0039] Example 2 A blood plasma substitute surface engineered bio-valve material is prepared through the following steps: Freshly collected porcine pericardium was washed with physiological saline at 4°C and 100 rpm for 1 hour to remove non-collagenous tissues such as fat. The pericardium was then immersed in a 70% ethanol aqueous solution containing 0.1 M 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) and activated at 37°C for 0.1 hours. 5% 2-aminoethyl methacrylate hydrochloride was further added, and the reaction was carried out at 37°C for 12 hours. The pericardium was then transferred to a solution containing 12% methacryloyl hydroxyethyl starch and soaked for 1 hour. Finally, 20 mM ammonium persulfate and 7 mM sodium bisulfite were added, and the reaction was carried out at 37°C for 8 hours. The pericardium was then washed with physiological saline to obtain the non-glutaraldehyde crosslinked plasma-modified bio-valve material.

[0040] In this embodiment, the methacrylated hydroxyethyl starch is prepared by the following steps: 10g of hydroxyethyl starch is dissolved in 100mL of deionized water, then 12mL of glycidyl methacrylate is added and reacted at room temperature for 48h. After dialyzing for 5d, the methacrylated hydroxyethyl starch is obtained by freeze drying.

[0041] It should be noted that the above embodiments are only some embodiments of the present invention. By changing the parameters such as the bio-valve material, the type and concentration of the amide condensing agent, the concentration of 2-aminoethyl methacrylate hydrochloride, the type and concentration of the initiator, the reaction temperature and time, etc., the preparation method described in the present invention can also successfully prepare the plasma substitute surface engineered bio-valve material of the present invention.

[0042] Comparative Example 1 A biological valve material is prepared through the following steps: Freshly collected pig pericardium was washed with physiological saline at 4℃ and 100 rpm for 1 hour to remove non-collagenous tissues such as fat. Then it was soaked in a 0.625% glutaraldehyde solution for 48 hours to obtain the bioprosthetic valve material.

[0043] Test Example 1: Cytotoxicity Test The toxicity of the bioprosthetic valve materials from Example 1 and Comparative Example 1 was evaluated using a co-culture assay with L929 cells and their extracts. Extracts of the bioprosthetic valve materials from Comparative Example 1 and Example 1 were prepared and used to replace the complete culture medium for culturing L929 cells. After 1 day of culture, viable cells in each group were stained with fluorescein diacetate (FDA) to observe their viability and number, reflecting the cytotoxicity of each group of materials. The staining results of L929 cells cultured with the extracts from Comparative Example 1 and Example 1 are shown below. Figure 2 As shown.

[0044] from Figure 2 As can be seen, compared with the glutaraldehyde-crosslinked biovalve material of Comparative Example 1, the biovalve material obtained in Example 1 of this invention has more surviving L929 cells after culturing the extract, which indicates that the biovalve material prepared by this invention has lower toxicity than the glutaraldehyde-crosslinked biovalve material.

[0045] Test Example 2: Blood Adhesion Test Fresh rabbit blood was used for blood adhesion experiments. The bioprosthetic valve materials obtained in Comparative Example 1 and Example 1 were cut into circular samples with a diameter of 10 mm and washed three times repeatedly with PBS buffer. The samples were then placed in 48-well plates, with 300 μL of rabbit blood added to each well, and incubated at 37°C on a shaker (70 rpm) for 1 hour. After incubation, each group of materials was washed three more times with PBS, then fixed with glutaraldehyde, dehydrated with graded ethanol, and sputter-coated with gold. The blood adhesion on the surface of the bioprosthetic valve materials was observed using scanning electron microscopy to reflect the antithrombotic properties of each group of bioprosthetic valve materials. The blood adhesion of the bioprosthetic valve materials obtained in Comparative Example 1 and Example 1 is shown in the scanning electron microscope images. Figure 3 As shown.

[0046] from Figure 3 As can be seen from the blood adhesion test, the bioprosthetic valve material prepared in Example 1 of this invention had fewer blood cells adhering to it than the bioprosthetic valve material prepared in Comparative Example 1 (glutaraldehyde cross-linked bioprosthetic valve material). This indicates that the bioprosthetic valve material prepared in this invention has better antithrombotic properties.

[0047] Test Example 3: Immune Response Test The bioprosthetic valve material samples prepared in Comparative Example 1 and Example 1 were cut into 1×1cm pieces. 2 Square-shaped samples were implanted into the subcutaneous tissue of rats. After 7 days, samples from each group were removed, fixed with paraformaldehyde, and then subjected to H&E staining to assess the degree of immune response induced by implantation. The degree of response was determined by observing the aggregation of immune cells around the samples. The H&E staining analysis results after implantation in Comparative Example 1 and Example 1 are as follows: Figure 4 As shown.

[0048] from Figure 4 As can be seen, compared with the bioprosthetic valve material prepared in Comparative Example 1, the bioprosthetic valve material prepared in Example 1 of this invention has fewer cell nuclei in the surrounding fibrous capsule tissue after staining, indicating that the number of immune cells recruited after implantation is significantly lower than that of the bioprosthetic valve material prepared in Comparative Example 1 (i.e., the bioprosthetic valve material treated with glutaraldehyde crosslinking), indicating that the level of immune response after implantation of the bioprosthetic valve material prepared in this invention is low.

[0049] Test Example 4: Calcification Test The bioprosthetic valve material samples prepared in Comparative Example 1 and Example 1 were cut into 1×1cm pieces. 2 Square-shaped bioprosthetic valve samples were implanted into the subcutaneous tissue of rats. Thirty days after implantation, samples from each group were removed, fixed with paraformaldehyde, and stained with alizarin red to assess the degree of calcification following implantation. The degree of calcification was determined by observing the intensity of the alizarin red staining on the bioprosthetic valve samples. The alizarin red staining analysis results after implantation in Comparative Example 1 and Example 1 are as follows: Figure 5 As shown.

[0050] from Figure 5 It can be seen that the color of the bio-valve material constructed in Example 1 of the present invention is significantly lighter after staining, indicating that its degree of calcification is significantly lower than that of the bio-valve material prepared in Comparative Example 1 (i.e., the bio-valve material treated with glutaraldehyde crosslinking). This shows that the bio-valve material prepared in the present invention has a lower tendency to calcify and exhibits better anti-calcification ability.

[0051] It should be noted that the above test results are only the test results of some embodiments of the present invention. The bioprosthetic valve material prepared by the preparation method described in the present invention has similar properties to that in Example 1, and both have lower toxicity, better antithrombotic properties, lower immune response levels, and better anti-calcification ability.

[0052] In summary, this invention enables the fabrication of plasma-modified bioprosthetic valve materials, which reduce the risk of calcification, overcome the thrombosis defects caused by xenogeneic collagen matrix in bioprosthetic valve materials, and also overcome the immune response caused by xenogeneic collagen matrix in bioprosthetic valve materials. Compared with existing technologies, this invention represents a significant advancement.

[0053] The above description is merely a representative embodiment of the present invention and is not intended to limit the present invention in any way. Any embodiment made by those skilled in the art without departing from the scope of the present invention and utilizing the disclosed technical content is an equivalent embodiment of the present invention. Any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A method for preparing a blood plasma substitute surface engineered bio-valve material, comprising the following steps: S1: Obtain bioprosthetic valve material and pretreat the bioprosthetic valve material; S2: The pretreated bioprosthetic valve material is activated with an amide condensing agent to obtain bioprosthetic valve material with activated carboxyl groups; S3: The activated carboxyl group of the biovalve material is grafted with 2-aminoethyl methacrylate hydrochloride to obtain a biovalve material grafted with methacrylate groups; S4: Under the action of an initiator, methacrylamide hydroxyethyl starch is polymerized with the bio-valve material grafted with methacrylate groups, and then washed to obtain the plasma substitute surface engineered bio-valve material.

2. The method for preparing engineered bio-valve material for blood plasma substitute surface according to claim 1, characterized in that, In step S1, the bioprosthetic valve material is any one of the following: pericardium, valve, intestinal membrane, meninges, pulmonary membrane, blood vessel, skin, amnion, and ligament.

3. The method for preparing engineered bio-valve material for blood plasma substitute surface according to claim 1, characterized in that, In step S1, the preprocessing includes the following sub-steps: S11: The obtained bioprosthetic valve material was stored in physiological saline under low temperature and humid conditions at 0-5℃; S12: Wash the bioprosthetic valve material preserved in step S11 with physiological saline on a shaker until there are no visible adhering non-pericardial or non-collagenous tissues and residues.

4. The method for preparing engineered bio-valve material for blood plasma substitute surface according to claim 1, characterized in that, In step S2, the amide condensing agent is a combination of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) or 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM).

5. The method for preparing engineered bio-valve material for plasma substitute surface according to claim 1 or 4, characterized in that, Step S2 specifically includes the following sub-steps: S21: Prepare an amide condensing agent solution; S22: Immerse the pretreated bio-valve material in the amide condensing agent solution to bring the amide condensing agent into contact with the bio-valve material to activate the carboxyl groups on the bio-valve material, thereby obtaining the bio-valve material with activated carboxyl groups.

6. The method for preparing the engineered bio-valve material for the surface of blood plasma substitute according to claim 5, characterized in that, In step S21, the concentration of the amide condensing agent solution is 0.05-0.5M; in step S22, the soaking time is 0.1-1h and the temperature is 4-40℃.

7. The method for preparing engineered bio-valve material for blood plasma substitute surface according to claim 1, characterized in that, Step S3 specifically includes the following sub-steps: adding 2-aminoethyl methacrylate hydrochloride to the solution of the bio-valve material soaked with the activated carboxyl group, and then reacting at 4-50℃ for 12-24h to obtain the bio-valve material grafted with the methacrylate group; Step S4 specifically includes the following sub-steps: S41: Prepare a solution of methacrylated hydroxyethyl starch; S42: The bio-valve material grafted with methacrylate groups is immersed in the methacryloyl hydroxyethyl starch solution, an initiator is added and the reaction is carried out for 5-24 hours, followed by washing to obtain the plasma substitute surface engineered bio-valve material.

8. The method for preparing the engineered bio-valve material for the surface of blood plasma substitute according to claim 7, characterized in that, In step S3, the mass concentration of the 2-aminoethyl methacrylate hydrochloride is 1%-5%; in step S41, the concentration of the methacrylated hydroxyethyl starch solution is 1%-20%.

9. The method for preparing engineered bio-valve material for plasma substitute surface according to claim 1 or 7, characterized in that, In step S4, the initiator is a redox initiator, and the oxidant of the redox initiator is any one or more of sodium persulfate, ammonium persulfate, and potassium persulfate; the reducing agent of the redox initiator is any one or more of sodium sulfite, sodium bisulfite, and potassium bisulfite; the concentration of the oxidant and the reducing agent is 2-50 mM.

10. A blood plasma substitute surface engineered bio-valve material, characterized in that, It is prepared using the preparation method of the plasma substitute surface engineered bio-valve material according to any one of claims 1-9.

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