A method of sheeting a biological valve material

Abstract

The application relates to the field of biological valve material processing, and particularly discloses a biological valve material sheeting method, which comprises the following steps: fresh bovine pericardium pretreatment to obtain bovine pericardium sheets; a fixing cloth soaked with a glutaraldehyde solution with an infiltration concentration of 0.28-0.32% is prepared; the bovine pericardium sheets are laid on the fixing cloth with the rough surfaces facing upwards and the smooth surfaces facing downwards, the bovine pericardium sheets and the fixing cloth are alternately sheeted, dry cloth is laid on the surface of the uppermost fixing cloth after exhaust, and fixed bovine pericardium sheets are obtained; then the bovine pericardium sheets are preliminarily treated by the glutaraldehyde solution with the concentration of 0.28-0.32%, and the storage temperature is 2-8 DEG C; then the preliminarily treated bovine pericardium sheets and the fixing cloth are packed, the bovine pericardium sheets are soaked in the glutaraldehyde solution with the concentration of 0.58-0.62% for secondary treatment, the storage temperature is 2-8 DEG C, then post-treatment and preservation are carried out, finally, the finished product is obtained by storage at 2-8 DEG C; and the finished product has the advantages of good durability, high stability and good calcification resistance.

Description

Technical Field

[0001] This application relates to the field of bioprosthetic valve material processing, and more specifically, it relates to a method for laying up bioprosthetic valve materials. Background Technology

[0002] An artificial heart valve is an implantable cardiac interventional medical device used to treat heart valve disease or defects.

[0003] Bioprosthetic valves are made from animal tissues (such as porcine valves or bovine pericardium), which are highly biocompatible with human tissues. After implantation, they are less likely to cause thrombosis or immune rejection. Furthermore, they do not require lifelong anticoagulation after surgery, significantly reducing the risk of bleeding. They are especially suitable for patients in urgent need of anticoagulation or elderly patients.

[0004] However, bioprosthetic valve materials are prone to calcification and degradation, with an average lifespan of about 10-15 years. Younger patients may need a second surgery to replace them. Furthermore, bioprosthetic valves are subjected to the pressure of heartbeats and blood flow in the human body for a long time, which can easily cause damage and affect the stability of the bioprosthetic valve.

[0005] Therefore, how to prepare a bioprosthetic valve material with good durability, high stability, good resistance to calcification, and resistance to protease hydrolysis is a problem that needs to be solved. Summary of the Invention

[0006] In order to prepare a bioprosthetic valve material with good durability, high stability, good resistance to calcification and resistance to protease hydrolysis, this application provides a method for laying up the bioprosthetic valve material.

[0007] This application provides a method for laying up a bioprosthetic valve material, which adopts the following technical solution:

[0008] A method for laying up a bioprosthetic valve material includes the following steps:

[0009] S1. Fresh beef heart slices are obtained after being cleaned, defat-removed, and decellularized.

[0010] S2. Prepare a fixation cloth impregnated with a 0.28-0.32% glutaraldehyde solution for later use;

[0011] S3. Lay the ox heart pericardium slice with the rough side up and the smooth side down on the fixing cloth. After degassing, repeat the above operation on the second ox heart pericardium slice, so that the ox heart pericardium slice and the fixing cloth are laid alternately. After degassing, lay a dry cloth on the surface of the top layer of fixing cloth to obtain a fixed ox heart pericardium slice.

[0012] S4. The fixed bovine heart pericardium tablets are initially treated with a 0.28-0.32% glutaraldehyde solution, with a storage temperature of 2-8℃. Then, the initially treated bovine heart pericardium tablets and the fixing cloth are packaged separately. The bovine heart pericardium tablets are then soaked in a 0.58-0.62% glutaraldehyde solution for a second treatment, with a storage temperature of 2-8℃, to obtain a semi-finished product.

[0013] S5. The semi-finished product undergoes post-processing and preservation, and is finally stored at 2-8℃ to obtain the finished product.

[0014] By adopting the above technical solution, glutaraldehyde is used to impregnate the fixation cloth. Glutaraldehyde molecules contain two active aldehyde groups, which can specifically condense with the free amino groups of bovine pericardial collagen in a neutral or weakly alkaline environment, thereby forming stable covalent bonds. Glutaraldehyde molecules can also establish intermolecular cross-linking networks between adjacent collagen fibers, improving the spatial stability of the fiber structure and transforming bovine pericardium from a flexible biological tissue into an engineering material with certain mechanical strength. The impregnated fixation cloth is laid flat in contact with the bovine pericardium slices. The cross-linking fixation effect of glutaraldehyde, combined with the binding of the fixation cloth, makes the cross-linked and fixed bovine pericardium slices relatively flat and have a certain strength.

[0015] Glutaraldehyde cross-links and fixes the enzymatic sites of collagen in bovine pericardial tissue, blocking the erosion pathway of proteases and extending the service life of bioprosthetic valve materials. Combined with the cross-linking effect of the three-dimensional network structure, it increases the cross-linking density of bioprosthetic valve materials, further improving their stability. It also has good resistance to calcification and protease hydrolysis, thus extending the service life of bioprosthetic valve materials.

[0016] Preferably, in step S2, glutaraldehyde solution is poured into the Lock & Lock box to immerse the fixing cloth.

[0017] By adopting the above technical solution, it is ensured that the fixation cloth is completely soaked in glutaraldehyde solution, and that bubbles are not easily generated. At the same time, it can also ensure the cross-linking and fixation effect of glutaraldehyde solution on bovine pericardial tissue, and ensure the cross-linking and fixation density, thereby giving the bioprosthetic valve material high strength and good stability.

[0018] Preferably, in step S3, the fixing cloth is gently squeezed to remove 60-70% of the liquid, fixed in the center of the bottom of the Lock & Lock box, and then the air bubbles are squeezed out. The surface liquid of the ox heart slice is removed, and after being stretched flat, the ox heart slice is laid flat on the fixing cloth with the rough side facing up and the smooth side facing down. After the air is released, the second ox heart slice is repeated in the same way, so that the ox heart slice and the fixing cloth are laid alternately. After the air is released, a dry cloth is laid on the surface of the top layer of fixing cloth to obtain the fixed ox heart slice.

[0019] By adopting the above technical solution, removing excess liquid can ensure the fixation and cross-linking stability and structural density of bovine pericardial tissue tablets. Furthermore, by venting, the structural density is further ensured, giving the bioprosthetic valve material good stability and durability.

[0020] Preferably, the dry cloth has 2-4 layers and is made of polyester fiber.

[0021] By employing the above technical solution, the overlapping bovine pericardial flaps and fixation flaps are sealed and the glutaraldehyde solution is stabilized. This not only prevents wrinkles from forming and ensures the directional alignment of collagen fibers, but also protects the bovine pericardial flaps from external damage. At the same time, it eliminates air bubbles, reduces structural defects formed after cross-linking and fixation, further ensures the stability of the cross-linked and fixed structure, and also blocks bacteria from the external environment, further extending the service life and durability of the bioprosthetic valve material.

[0022] Preferably, the total height of the ox heart bag sheet, fixing cloth, and dry cloth after they are laid out is less than 2 / 3 of the Lock & Lock box.

[0023] By adopting the above technical solution, a 1 / 3 space is reserved to ensure that the glutaraldehyde solution completely submerges the slice structure, preventing uneven cross-linking inside the pericardial slice due to insufficient liquid level.

[0024] Preferably, the specific steps of post-processing in S5 are as follows:

[0025] The semi-finished product is first fixed in a polylysine composite solution, then fixed a second time in a magnesium chloride composite solution, washed, and then soaked in a glutaraldehyde solution.

[0026] By adopting the above technical solution, after the semi-finished product is treated with glutaraldehyde solution, residual aldehyde groups and other substances remain. The residual unreacted glutaraldehyde monomers and free aldehyde groups become the core sites for calcification. The amino groups of polylysine in the polylysine composite solution actively bind to the residual aldehyde groups to form stable covalent bonds, reducing the nucleation sites for calcification. In addition, the dense cationic layer formed by polylysine blocks the diffusion path of calcium ions, further preventing calcification. The carboxyl groups of polylysine can enhance the hydrophilicity of the material surface and can also bind to calcium ions to form stable complexes, reducing the free concentration of calcium ions in tissues, reducing calcium salt deposition, and improving the anti-calcification effect of the bio-valve material. Finally, the magnesium chloride composite solution is used for treatment. After the polylysine blocks the aldehyde groups, the magnesium chloride further forms a barrier, constructing a double anti-calcification defense line and improving the anti-calcification effect.

[0027] The combination of polylysine composite solution and magnesium chloride composite solution utilizes the linear polypeptide chains of polylysine to penetrate into the interstitial spaces of bovine pericardial collagen fibers, further binding with the carboxyl groups of internal collagen to reduce porosity and increase structural density. Furthermore, the amino groups in polylysine form covalent bonds with the residual aldehyde groups of glutaraldehyde, adding molecular bridging points to the original cross-linked network and further increasing the cross-linking density. Meanwhile, magnesium ions in magnesium chloride bind with carboxyl groups to further fill the pores, increasing the density of the cross-linked structure. This combination enhances the strength and stability of the bioprosthetic valve material, extending its service life and durability.

[0028] Preferably, the polylysine composite solution is composed of a polylysine solution and a phosphatidylserine solution in a mass ratio of 1:0.1-0.2. During the secondary crosslinking process, the polylysine solution is added first, followed by the addition of the phosphatidylserine solution.

[0029] By employing the above technical solution, polylysine solution and phosphatidylserine solution are combined. The residual aldehyde groups after glutaraldehyde cross-linking by polylysine block the calcium ion binding anchor points. Furthermore, the polypeptide chains of polylysine further cross-link with phosphatidylserine to form a cross-linked network layer. The hydrophilic end of phosphatidylserine is cross-linked with polylysine, while the hydrophobic end isolates calcium ions from collagen, inhibiting the formation of hydroxyapatite crystal nuclei and improving anti-calcification and anti-protease hydrolysis effects. The formation of the cross-linked network further enhances structural stability. Moreover, polylysine and phosphatidylserine can bridge the carboxyl groups of adjacent collagen fibers, reducing inter-fiber slippage and further improving the strength and fracture stress of the bioprosthetic valve material, thereby enhancing its strength and stability.

[0030] Preferably, the magnesium chloride composite solution is composed of a magnesium chloride solution and a polyvinyl alcohol-1799 solution with a mass ratio of 1:0.4-0.6.

[0031] By employing the above technical solutions, magnesium ions compete with calcium ions to bind to residual carboxyl groups and glutaraldehyde aldehyde groups, blocking calcification initiation sites. Combined with the hydrophilicity of the dense hydroxyl groups in the polyethylene glycol-1799 molecular chain, this further inhibits calcium crystallization nucleation. After magnesium ions adsorb onto the collagen surface, polyvinyl alcohol-1799 connects with polylysine and phosphatidylserine to form a barrier, preventing calcium deposition and improving the anti-protease hydrolysis effect. Furthermore, magnesium ions bind adjacent collagen fibers, reducing fiber slippage and improving the strength and elasticity of the bioprosthetic valve material. This, combined with the barrier network formed by polyvinyl alcohol-1799, further enhances the strength and durability of the bioprosthetic valve material.

[0032] Preferably, the fixing fabric is made of polyester fiber fabric.

[0033] By adopting the above technical solution, when the moist bovine pericardium patch is spread on the surface of the polyester fiber cloth, the tensile strength of the polyester fiber cloth can absorb the local shear force generated by the spreading operation, reduce the risk of collagen microfiber breakage, and further ensure the spreading stability of the bioprosthetic valve material.

[0034] Preferably, the polyester fiber cloth is treated as follows: the pretreated polyester fiber cloth is soaked in physiological saline, taken out and dried, then placed in polyethylene glycol-silver powder composite solution for immersion and dispersion treatment, then the pretreated polyester fiber cloth is taken out and dried to obtain polyester fiber cloth.

[0035] By adopting the above technical solution, the bonding effect between polyethylene glycol and glutaraldehyde is utilized to ensure that glutaraldehyde is evenly penetrated between each layer. Combined with the antibacterial effect of silver powder, the bioprosthetic valve material is not easily affected by bacteria. Furthermore, the binding effect of polyethylene glycol on the surface of the pretreated polyester fiber cloth allows the silver powder to be stably attached to the surface of the polyester fiber cloth, while also helping to expel air bubbles, thus completing the laying of the bioprosthetic valve material.

[0036] In summary, this application has the following beneficial effects:

[0037] 1. The bovine pericardial patch used as a bioprosthetic valve material has a smooth appearance, uniform thickness, and is free of air bubbles;

[0038] 2. The bovine pericardial patch, a bioprosthetic valve material, has stable mechanical properties and is not easily deformed;

[0039] 3. The bovine pericardial flap material for biological valves has excellent thermodynamic properties, with a thermal shrinkage temperature exceeding 85℃. Attached Figure Description

[0040] Figure 1 This is a schematic diagram of the stretched and fixed cloth in this application;

[0041] Figure 2 This is a schematic diagram of the installation of the fixing cloth in this application;

[0042] Figure 3 This is a schematic diagram of the calf pericardium sheet laid out in this application;

[0043] Figure 4 This is a schematic diagram of the beef heart slices in the Lock & Lock box after being packaged according to this application. Detailed Implementation

[0044] The present application will be further described in detail below with reference to the embodiments.

[0045] Preparation example of polylysine composite solution

[0046] Of the following raw materials, polylysine was purchased from Shandong Xinxiong Biotechnology Co., Ltd. as food-grade ε-polylysine; phosphatidylserine was purchased from Hebei Qiansheng Biotechnology Co., Ltd.; and all other raw materials were commercially available.

[0047] Preparation Example 1: The polylysine composite solution was prepared using the following method:

[0048] Weigh out polylysine and place it in water, stirring until completely dissolved to obtain a 1% polylysine solution.

[0049] Phosphatidylserine was placed in isopropanol and stirred until completely dissolved to obtain a 1% (w / w) phosphatidylserine solution; the isopropanol concentration was 95%.

[0050] Example of preparation of magnesium chloride composite solution

[0051] All of the following ingredients are commercially available.

[0052] Preparation Example 2: Magnesium chloride composite solution was prepared using the following method:

[0053] Magnesium chloride was placed in water and stirred until completely dissolved to obtain a 1% magnesium chloride solution. Polyvinyl alcohol-1799 was placed in hot water at 95°C and stirred until completely dissolved to obtain a 1% polyvinyl alcohol-1799 solution. 1 kg of magnesium chloride solution and 0.5 kg of polyvinyl alcohol-1799 solution were mixed and stirred evenly to obtain a magnesium chloride composite solution.

[0054] Preparation Example 3: The difference between this preparation example and Preparation Example 2 is that:

[0055] Mix 1 kg of magnesium chloride solution and 0.4 kg of polyvinyl alcohol-1799 solution until homogeneous to obtain magnesium chloride composite solution.

[0056] Preparation Example 4: The difference between this preparation example and Preparation Example 2 is that:

[0057] Mix 1 kg of magnesium chloride solution and 0.6 kg of polyvinyl alcohol-1799 solution until homogeneous to obtain magnesium chloride composite solution.

[0058] Example of polyester fiber fabric preparation

[0059] The pretreated polyester fiber cloth used in the following raw materials was purchased from Hejian Qingfeng Asbestos Chemical Co., Ltd., and was 18-gauge; the other raw materials were all commercially available.

[0060] Preparation Example 5: Polyester fiber fabric was prepared using the following method:

[0061] Polyethylene glycol was placed in water and stirred until completely dissolved. The polyethylene glycol was polyethylene glycol 2000, resulting in a 2% (w / w) polyethylene glycol solution. 0.2 kg of silver powder (100 nm) was dispersed in 1 kg of the polyethylene glycol solution. The dispersion was performed by ultrasonic dispersion at a frequency of 20 kHz for 5 min. After mixing, a polyethylene glycol-silver powder composite solution was obtained.

[0062] The pretreated polyester fiber was soaked in physiological saline for 10 minutes, then removed and dried. It was then soaked in a polyvinyl alcohol-silver powder composite solution for 10 minutes. During the soaking process, it was sonicated at 20 kHz for 5 minutes. The pretreated polyester fiber was then removed and dried to obtain the polyester fiber fabric.

[0063] Example

[0064] All of the following ingredients are commercially available.

[0065] Example 1: A method for laying up a bioprosthetic valve material:

[0066] S1. Wash fresh bovine pericardium with clean water, remove excess adipose tissue, cut off a relatively uniform portion of the precordial region, and store in ice-cold physiological saline. Then, add 0.01M Tris-HCl (pH 8.0) buffer containing 0.2g EDTA and 100μg PMSF at a ratio of 1:4 (weight:volume), shake at 4°C for 4 hours at a shaking frequency of 120 rpm, discard the liquid, keep the bovine pericardium, and wash it 3 times with physiological saline. Then, add 0.05M Tris-HCl (pH 8.0) buffer containing 0.5% Triton X-100, 1% deoxycholic acid, 0.2g EDTA, and 100μg PMSF at a ratio of 1:4 (weight:volume), shake at 4°C for 4 hours at a shaking frequency of 120 rpm, discard the liquid, keep the bovine pericardium, and wash it 3 times with physiological saline to complete the decellularization process, obtaining bovine pericardium slices for later use. Store in a medical refrigerator.

[0067] S2. Take a Lock & Lock box, put the fixing cloth into the Lock & Lock box, pour 0.3% glutaraldehyde solution into the Lock & Lock box to immerse the fixing cloth, so that the fixing cloth is fully soaked in the 0.3% glutaraldehyde solution, and set aside; the fixing cloth is polyester fiber cloth.

[0068] S3. Take a Lock & Lock box as the container for cross-linking and fixing the bovine heart capsule tablets. Two production workers each take a piece of fixing cloth soaked in 0.3% glutaraldehyde solution, gently squeeze the fixing cloth to remove 60% of the liquid, keeping the fixing cloth moist. Each worker stretches up one side and two corners of the fixing cloth, and together they stretch the fixing cloth flat and lay it in the center of the bottom of the Lock & Lock box. If air bubbles are generated, they should be expelled. The surface of the fixing cloth should be flat and free from wrinkles, overlaps, or creases. A total of three layers of fixing cloth are laid (see...). Figure 1 and Figure 2 After wiping off the liquid from the surface of the pericarp slice in S1, two people each stretch two corners of the pericarp slice and work together to slowly flatten it, laying it flat on the fixing cloth with the rough side facing up and the smooth side facing down, so that the pericarp slice is completely attached to the center of the fixing cloth (see...). Figure 3During this process, tension must not be used to pull the pericarp sheet. If air bubbles are generated, they should be removed. The surface of the pericarp sheet should be free of artificial wrinkles, folds, air bubbles, etc., to complete the laying of the first pericarp sheet. The laying of the second pericarp sheet is carried out in the same way as above, with the pericarp sheet and fixing cloth being laid alternately. After all pericarp sheets are laid, the top layer is a dry cloth made of polyester fiber, laid in 3 layers. During the laying process, the pericarp sheet and fixing cloth should not have wrinkles, folds, air bubbles, etc. that affect the flatness of the pericarp sheet surface. The total height after the pericarp sheet and fixing cloth are laid alternately should be less than two-thirds of the height of the Lock & Lock box to obtain the fixed pericarp sheet. Once this height is reached, another Lock & Lock box is used for laying the sheets again.

[0069] S4. One person gently presses down on the material inside the Lock & Lock box with their palm, while another person uses a 1000mL graduated cylinder filled with 0.3% glutaraldehyde solution to pour the solution into the Lock & Lock box containing the pericardium tablets. The solution is poured slowly along the inner wall of the Lock & Lock box, ensuring it fully soaks into the pericardium tablets and the fixing cloth. The volume of the 0.3% glutaraldehyde solution poured should be level with the height of the top layer of fixing cloth inside the Lock & Lock box. During this process, avoid floating or shifting the pericardium tablets and fixing cloth. Cover and label the box, then place it in a medical refrigerator at 4°C for 48 hours to complete the initial treatment. Remove the Lock & Lock box from the medical refrigerator in the general production area after the initial 0.3% glutaraldehyde solution treatment. Wipe the surface of the Lock & Lock box clean with a lint-free cloth, then transfer it through a transfer window to the Class 10,000 production area. After cleaning the surface with 75% alcohol, open the Lock & Lock container and transfer the securing cloth and pericarp pieces one by one from top to bottom. Place the transferred pericarp pieces and securing cloth in two separate Lock & Lock containers. The pericarp pieces should be laid flat and stacked in the center of the Lock & Lock containers in sequence, maintaining their original shape during the transfer process. Do not fold, tear, or otherwise damage the pericarp's shape. After collecting the securing cloth, wash, dry, and store it according to regulations. One person gently presses the top of the pericarp pieces in the Lock & Lock container with their palm, while another person uses a 1000mL graduated cylinder filled with 0.6% glutaraldehyde solution to slowly pour the solution into the Lock & Lock container containing the transferred pericarp pieces. The solution should be poured slowly along the inner wall of the Lock & Lock container, ensuring the top pericarp is submerged (see [link to relevant documentation]). Figure 4 The bovine pericardium slices were placed in a medical refrigerator at a storage temperature of 4°C for 7 days to complete the secondary processing. During the process, it was important to control any phenomena that would damage the surface morphology of the bovine pericardium slices, such as folding or creasing, to obtain a semi-finished product.

[0070] S5. Take out the bovine heart capsules one by one from top to bottom and place them into a new Lock & Lock box. The bovine heart capsules should be laid flat and stacked in the center of the Lock & Lock box in order. Store each box with 20 bovine heart capsules. If there are fewer than 20, store each box with the actual number of capsules. One person gently presses the top of the bovine heart capsules in the Lock & Lock box after the previous step with their palm. Another person uses a 500mL graduated cylinder filled with 0.3% glutaraldehyde solution to pour 0.3% glutaraldehyde solution into the Lock & Lock box containing the transferred bovine heart capsules. The solution should be poured slowly along the inner wall of the Lock & Lock box. The volume of the 0.3% glutaraldehyde solution should be enough to submerge the top layer of bovine heart capsules. Finally, close the lid and store in a medical refrigerator at a storage temperature of 4°C to obtain the finished product. The thickness of the finished valve material is 0.5mm.

[0071] Example 2: The difference between this example and Example 1 is that:

[0072] The fixing fabric used is the fixing fabric prepared in Preparation Example 5;

[0073] The storage temperature during the preliminary and secondary processing in S4 is 4℃;

[0074] S5. The semi-finished product is first immersed in a polylysine composite solution at 35°C for 60 minutes, then phosphatidylserine solution is added and the immersion continues for 30 minutes. The polylysine solution and phosphatidylserine solution together form the polylysine composite solution. The polylysine solution and phosphatidylserine solution used are those prepared in Preparation Example 1, with a mass ratio of 1:0.15. This is used for fixation. Then, it is immersed in a magnesium chloride composite solution prepared in Preparation Example 2 at 35°C for 30 minutes to complete the secondary fixation treatment. After washing three times with physiological saline, it is then... Remove the veal slices one by one from top to bottom and place them into a new Lock & Lock box. The veal slices should be laid flat and stacked in the center of the Lock & Lock box in order. Store each box with 20 veal slices. If there are fewer than 20 slices, store each box with the actual number of slices. One person gently presses down on the top of the veal slices in the Lock & Lock box after the previous step with their palm. Another person uses a 500mL graduated cylinder filled with 0.3% glutaraldehyde solution to pour 0.3% glutaraldehyde solution into the Lock & Lock box containing the transferred veal slices. The solution should be poured slowly along the inner wall of the Lock & Lock box. The volume of the 0.3% glutaraldehyde solution should be enough to submerge the top layer of veal slices. Finally, close the lid and store in a medical refrigerator at a temperature of 4°C to obtain the finished product.

[0075] Example 3: The difference between this example and Example 2 is that:

[0076] In S2, the 0.3% glutaraldehyde solution is replaced with a 0.28% glutaraldehyde solution;

[0077] S3: The fixed cloth is gently squeezed to remove 60% of the liquid; the dry cloth consists of 2 layers.

[0078] In S4, the 0.3% glutaraldehyde solution was replaced with a 0.28% glutaraldehyde solution; the 0.6% glutaraldehyde solution was replaced with a 0.58% glutaraldehyde solution; the storage temperature during the preliminary and secondary treatments was 8℃.

[0079] The mass ratio of polylysine solution to phosphatidylserine solution in S5 is 1:0.1; the calcium chloride composite solution used is the calcium chloride composite solution prepared in Preparation Example 3.

[0080] Example 4: The difference between this example and Example 2 is that:

[0081] In S2, the 0.3% glutaraldehyde solution is replaced with a 0.32% glutaraldehyde solution;

[0082] S3: The fixed cloth is gently squeezed to remove 70% of the liquid; the dry cloth consists of 4 layers.

[0083] In S4, the 0.3% glutaraldehyde solution was replaced with a 0.32% glutaraldehyde solution; the 0.62% glutaraldehyde solution was replaced with a 0.58% glutaraldehyde solution; the storage temperature during the preliminary and secondary treatments was 2℃.

[0084] The mass ratio of polylysine solution to phosphatidylserine solution in S5 is 1:0.2; the calcium chloride composite solution used is the calcium chloride composite solution prepared in Preparation Example 4.

[0085] Example 5: The difference between this example and Example 2 is that:

[0086] In S5, the phosphatidylserine solution is replaced with an equal mass of polylysine solution.

[0087] Example 6: The difference between this example and Example 2 is that:

[0088] S5 did not undergo secondary fixation treatment with magnesium chloride composite solution.

[0089] Example 7: The difference between this example and Example 2 is that:

[0090] In S5, the magnesium chloride composite solution raw material is replaced with an equal mass of magnesium chloride solution to replace the polyvinyl alcohol-1799 solution.

[0091] Example 8: The difference between this example and Example 2 is that:

[0092] The polyester fiber fabric is the ordinary commercially available polyester fiber fabric.

[0093] Comparative Example

[0094] Comparative Example 1: The difference between this preparation example and Example 1 is that:

[0095] In S4 and S5, the storage temperature has been changed from 4°C to 25°C.

[0096] Comparative Example 2: The difference between this preparation example and Example 1 is that:

[0097] S2. Place the ox heart slices on a flat plate (polycarbonate plate), fix the four corners at four points, and then immerse them in a 0.3% glutaraldehyde solution for 48 hours. Then remove the ox heart slices, store them in boxes of 20, and obtain the finished product.

[0098] Performance testing

[0099] 1. Mechanical property testing

[0100] Biological valve materials were prepared using the methods of Examples 1-8 and Comparative Examples 1-2, respectively. The elongation, tensile strength, elastic modulus, and thermal shrinkage temperature were tested and recorded in accordance with the general requirements for animal-derived pericardial valve leaflets in T / CSBM0031-2023.

[0101] 2. Anti-calcification test

[0102] Biological valve materials were prepared using the methods described in Examples 1-2 and 5-7, respectively. The calcium content was measured and the data were recorded in the subcutaneous implantation experiment of rats for evaluating the anti-calcification of animal-derived cardiovascular implants, in accordance with YY / T1859-2022.

[0103] Table 1 Performance Test Table (In the table, " / " indicates that the corresponding embodiment or comparative example did not test this item, so there is no data)

[0104]

[0105]

[0106] As can be seen from Examples 1-4 and Table 1, the biomembrane material prepared in this application has high strength, good mechanical properties and good stability, and can extend the service life of biomolecular valve materials.

[0107] Combining Examples 2 and 5-8 with Table 1, it can be seen that in Example 5, replacing the phosphatidylserine solution with the same mass of polylysine solution resulted in lower fracture load and fracture strength compared to Example 2. This indicates that the addition of phosphatidylserine can increase the crosslinking density of the bioprosthetic valve material, thereby improving the strength, anti-calcification ability, and stability of the bioprosthetic valve material.

[0108] In Example 6S5, no secondary fixation treatment with magnesium chloride composite solution was added. Compared with Example 2, the fracture load and fracture strength of Example 6 were lower than the corresponding data of Example 2. This indicates that the magnesium chloride composite solution fixes and seals the bioprosthetic valve material, further improving the strength and mechanical properties of the bioprosthetic valve material, extending the service life and durability of the bioprosthetic valve material, and also improving the anti-calcification ability of the bioprosthetic valve material.

[0109] In Example 7S5, the magnesium chloride composite liquid raw material was replaced with an equal mass of magnesium chloride solution to replace the polyvinyl alcohol-1799 solution. Compared with Example 2, the fracture load and fracture strength of Example 7 were lower than the corresponding data of Example 7, indicating that polyvinyl alcohol-1799 is linked with polylysine and phosphatidylserine to form a barrier, which further improves the strength and durability of the bioprosthetic valve material and can also improve the anti-calcification ability of the bioprosthetic valve material.

[0110] Example 8 uses commercially available polyester fiber fabric. Compared to Example 2, the breaking load and breaking strength of Example 8 are lower than the corresponding data of Example 2. This indicates that by utilizing the bonding effect of polyethylene glycol and glutaraldehyde, the cross-linking and fixing effect of the sheet is further improved, thereby improving the strength and stability of the bioprosthetic valve material.

[0111] As can be seen from Example 1 and Comparative Example 1, and Table 1, cross-linking treatment of bovine pericardial tablets at room temperature affects their strength.

[0112] As can be seen from Example 1 and Comparative Example 2, and in conjunction with Table 1, fixing the bovine pericardium slice with a hard steel plate affects its flexibility and strength.

[0113] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A method of sheeting a biological valve material, characterized by, It comprises the following steps: S1, after washing, removing fat and cell treatment, fresh bovine pericardium is obtained, and is prepared for use; S2, a fixed cloth with glutaraldehyde solution of 0.28-0.32% infiltration concentration is prepared, and is prepared for use; S3, the rough surface of the bovine pericardium sheet is upward, and the smooth surface is downward, and is laid on the fixed cloth, and after exhausting, the second bovine pericardium sheet is repeated according to the above method, and the bovine pericardium sheet and the fixed cloth are alternately laid, and after exhausting, the dry cloth is laid on the surface of the uppermost fixed cloth, and the fixed bovine pericardium sheet is obtained; S4, the fixed bovine pericardium sheet is preliminarily treated by 0.28-0.32% glutaraldehyde solution, and the storage temperature is 2-8℃; then the preliminarily treated bovine pericardium sheet and the fixed cloth are packed, the bovine pericardium sheet is treated by 0.58-0.62% glutaraldehyde solution, and the storage temperature is 2-8℃, and the semi-finished product is obtained; S5, the semi-finished product is treated and stored, and finally stored at 2-8℃ to obtain the finished product; The specific steps of the post-treatment are as follows: The semi-finished product is first fixed by polylysine complex solution, and then treated by magnesium chloride complex solution, and after washing, it is soaked in glutaraldehyde solution; the polylysine complex solution is composed of polylysine solution and phosphatidylserine solution with a mass ratio of 1:0.1-0.2, and during the fixing process, the polylysine solution is added first, and then the phosphatidylserine solution is added; The magnesium chloride complex solution is composed of magnesium chloride solution and polyvinyl alcohol-1799 solution with a mass ratio of 1:0.4-0.

6.

2. The method of claim 1, wherein: In S2, the glutaraldehyde solution is poured into the buckle box, and the fixed cloth is immersed.

3. The method of claim 1, wherein the bioprosthetic valve material is a porcine pericardium. In S3, the fixed cloth is lightly squeezed to remove 60-70% of the liquid, and is fixed at the center of the bottom of the buckle box, then the bubbles are squeezed out, the surface liquid of the bovine pericardium sheet is removed, and after being flattened, the bovine pericardium sheet is laid on the fixed cloth with the rough surface upward and the smooth surface downward, after exhausting, the second bovine pericardium sheet is repeated according to the above method, and the bovine pericardium sheet and the fixed cloth are alternately laid, after exhausting, the dry cloth is laid on the surface of the uppermost fixed cloth, and the fixed bovine pericardium sheet is obtained.

4. The method of claim 3, wherein the biological valve material is a porcine pericardium. The dry cloth is 2-4 layers, and the dry cloth is polyester fiber cloth.

5. The method of claim 4, wherein the bioprosthetic valve material is a porcine pericardium. After the bovine pericardium sheet, the fixed cloth and the dry cloth are laid, the total height is less than 2 / 3 of the buckle box.

6. The method of claim 1, wherein the bioprosthetic valve material is a porcine pericardium. The fixed cloth is made of polyester fiber cloth.

7. The method of claim 6, wherein the bioprosthetic valve material is a porcine pericardium. The polyester fiber cloth is treated as follows: the pretreated polyester fiber cloth is soaked in physiological saline, dried after taking out, then soaked and dispersed in polyethylene glycol-silver powder complex solution, and then the pretreated polyester fiber cloth is taken out and dried to obtain the polyester fiber cloth.

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