CN-122005938-A - Preparation method of biological valve

CN122005938ACN 122005938 ACN122005938 ACN 122005938ACN-122005938-A

Abstract

The application provides a preparation method of a biological valve, which comprises the following steps of obtaining the biological film, carrying out first cross-linking on the biological film by using free enzyme to obtain the first cross-linked biological film, carrying out second cross-linking on the first cross-linked biological film by using glutaraldehyde to obtain the second cross-linked biological film, carrying out attenuation modification on the second cross-linked biological film by using free enzyme to obtain the attenuated modified biological film, and washing, dehydrating and drying to obtain the biological valve. The application successfully realizes the dual targets of 'attenuation' and 'enhancement' in the preparation of the biological valve through an innovative three-step combined crosslinking strategy, namely, the application completely maintains and even improves the key physical and chemical properties and structural stability of the valve while greatly reducing the potential cytotoxicity and immunogenicity, and provides a high-efficiency and reliable solution for preparing a safer and more durable new-generation dry biological valve.

Inventors

HU YANGYANG
SHANG DAPENG
WEN XIANTAO
YU QIFENG

Assignees

上海纽脉医疗科技股份有限公司

Dates

Publication Date: 20260512
Application Date: 20260227

Claims (9)

1. A method of preparing a biological valve comprising the steps of: Obtaining a biological film; performing primary crosslinking on the biological film by using free enzyme to obtain a primary crosslinked biological film; Performing secondary crosslinking on the primary crosslinked biomembrane by using glutaraldehyde to obtain a secondary crosslinked biomembrane; performing attenuation modification on the biological film after the second cross-linking by using free enzyme to obtain an attenuated modified biological film; Washing, dehydrating and drying to obtain the biological valve.
2. The preparation method according to claim 1, wherein, The free enzyme is at least one selected from transglutaminase, lysyl oxidase, tyrosinase, peroxidase and laccase.
3. The preparation method according to claim 2, wherein, The free enzyme is transglutaminase.
4. The preparation method according to claim 1, wherein, The mass fraction of free enzyme used for the first crosslinking is 1% -10%, or, The mass fraction of free enzyme used for attenuation modification is 1% -10%.
5. The preparation method according to claim 1, wherein, The mass fraction of the glutaraldehyde solution is 0.05% -0.25%.
6. The preparation method according to claim 1, wherein, The first crosslinking is carried out at a temperature of 20-45 ℃, a pH value of 5-9 and a reaction time of 24-72 hours.
7. The preparation method according to claim 1, wherein, The second crosslinking is carried out at a temperature of 10-40 ℃, a pH value of 7.3-8.5 and a reaction time of 48-120 hours.
8. The preparation method according to claim 1, wherein, The conditions of attenuation and modification are that the temperature is 20-45 ℃, the pH value is 5-9, and the reaction time is 24-72 hours.
9. The preparation method according to claim 1, wherein, The biological membrane is pig pericardium or cattle pericardium.

Description

Preparation method of biological valve Technical Field The application relates to a preparation method of a biological valve. Background Prosthetic heart valves are currently most commonly available on the market in both mechanical and biological valves, with most prosthetic biological valves being made from porcine or bovine pericardium as a raw material by glutaraldehyde crosslinking. Since glutaraldehyde is not fully crosslinked with the biological valve, an excess of glutaraldehyde is typically used to ensure the crosslinking effect. Meanwhile, glutaraldehyde solution is generally used as preservation solution for biological valves. These factors can cause an increase in residual aldehyde groups and can cause effects in terms of thrombosis, immune response, calcification, etc. after valve implantation, thereby causing a decrease in the overall effectiveness of the valve. Disclosure of Invention The application aims to solve the technical problems and provides a dry biological valve attenuation crosslinking method. The biological membrane is firstly crosslinked by enzyme, secondly crosslinked by low-concentration glutaraldehyde, then added with free enzyme again, attenuated and modified, and finally dehydrated and dried, so that the residual toxicity of glutaraldehyde is reduced as a whole, the crosslinking degree of the biological valve is improved, and the service life of the biological valve is prolonged. The technical scheme of the application is as follows: 1. a method of preparing a biological valve comprising the steps of: Obtaining a biological film; performing primary crosslinking on the biological film by using free enzyme to obtain a primary crosslinked biological film; Performing secondary crosslinking on the primary crosslinked biomembrane by using glutaraldehyde to obtain a secondary crosslinked biomembrane; performing attenuation modification on the biological film after the second cross-linking by using free enzyme to obtain an attenuated modified biological film; Washing, dehydrating and drying to obtain the biological valve. 2. The production method according to item 1, wherein, The free enzyme is at least one selected from transglutaminase, lysyl oxidase, tyrosinase, peroxidase and laccase. 3. The production method according to item 2, wherein, The free enzyme is transglutaminase. 4. The production method according to item 1, wherein, The mass fraction of free enzyme used for the first crosslinking is 1% -10%, or, The mass fraction of free enzyme used for attenuation modification is 1% -10%. 5. The production method according to item 1, wherein, The mass fraction of the glutaraldehyde solution is 0.05% -0.25%. 6. The production method according to item 1, wherein, The first crosslinking is carried out at a temperature of 20-45 ℃, a pH value of 5-9 and a reaction time of 24-72 hours. 7. The production method according to item 1, wherein, The second crosslinking is carried out at a temperature of 10-40 ℃, a pH value of 7.3-8.5 and a reaction time of 48-120 hours. 8. The production method according to item 1, wherein, The conditions of attenuation and modification are that the temperature is 20-45 ℃, the pH value is 5-9, and the reaction time is 24-72 hours. 9. The production method according to item 1, wherein, The biological membrane is pig pericardium or cattle pericardium. The low-concentration glutaraldehyde and enzyme combined attenuation crosslinking method has the aldehyde group content which is obviously lower than that of the traditional glutaraldehyde crosslinking method, and the mechanical strength and the thermal shrinkage temperature are equivalent to those of the traditional glutaraldehyde crosslinking method, so that the attenuation crosslinking method can reduce the aldehyde group content while ensuring the crosslinking strength, and effectively reduce the potential risk brought by residual aldehyde groups in the traditional glutaraldehyde crosslinking method. The preparation method of the biological valve provided by the application has the following outstanding technical effects: The method has the advantages of high efficiency and toxicity reduction, and improves the biocompatibility, and obviously reduces the content of residual glutaraldehyde (aldehyde group) in the final product through the synergistic process of primary cross-linking of free enzyme, secondary cross-linking of low-concentration glutaraldehyde and toxicity reduction modification of free enzyme. The method is expected to directly reduce the risks of thrombosis, immune inflammatory reaction, tissue calcification and the like caused by aldehyde residue after valve implantation, thereby remarkably improving the biocompatibility and long-term safety of the biological valve. The mechanical property and the stability are excellent, and on the premise of greatly reducing the dosage of glutaraldehyde, the mechanical strength (tensile strength) and the thermal stability (thermal shrinkage temperature) of the b