Cross-linked artificial biological valve and preparation method thereof
Technical Field
The invention relates to the field of medical instruments, in particular to a cross-linked artificial biological valve and a preparation method thereof.
Background
With economic development and aging population, the incidence of senile Calcified Aortic Valve Disease (CAVD) is on the rise, second to coronary heart disease and hypertension. A retrospective non-stochastic research analysis in China suggests that the incidence of Aortic Valve Calcification (AVC) in middle-aged and elderly patients over 50 years old reaches 49.38%. With the aging of population, the incidence of Calcified Aortic Stenosis (CAS) is increasing, which is about to become the leading cause of valvular disease in our country.
Aortic Valve Replacement (AVR) using prosthetic valves is the gold standard for treating various types of aortic valve lesions. The use of transcatheter aortic biologic valve placement (TAVR, or TAVI) is increasing worldwide. This is probably because this technique has the potential to reduce mortality and morbidity compared to the high risk of traditional surgical AVR. Transcatheter aortic valve implantation offers new promise as a less invasive, less risky treatment for patients with severe aortic stenosis, particularly those who cannot undergo open thoracic surgery. With the wide use of the interventional artificial heart biological valve, how to improve the service life of the interventional artificial heart biological valve and reduce the decay of the biological valve are important problems facing people.
The existing biological valve is basically crosslinked by adopting a single glutaraldehyde solution, the mechanical property and durability of the use range are achieved under certain conditions, the biological valve has good biocompatibility, and then the biological valve is sewn on a memory metal bracket and then stored in glutaraldehyde storage solution with certain concentration. Prior to heart valve replacement surgery, the valve is transported to a hospital operating room, cleaned, and then delivered into the patient's heart by a delivery device. During the manufacturing, sewing, transporting and using processes of the valve, the valve must be preserved in a specific preservation solution and the valve is ensured to be in a wet state.
The existing crosslinked biological valve at home and abroad needs to be stored in a specific storage solution and ensures that the valve is in a wet state in the manufacturing, sewing, transporting and using processes, so that the valve needs additional conditions in the preparation process so as to be complicated in the preparation process, the valve needs to be separated from a conveyor in the transporting process, the valve needs to be loaded before an operation and needs to be cleaned from the residual storage solution on the valve when being loaded, the risk of calcification-caused inactivation and bacteria infection is easily lost, and the clinical application and popularization of the valve are limited. Increasing the cost of biological valve storage, transport, and use. Moreover, after the biological valve is cross-linked by glutaraldehyde, unpaired aldehyde groups from the glutaraldehyde and carboxyl groups on the valve protein tissue often remain on the valve, and the aldehyde groups and the carboxyl groups are easy to combine with calcium ions, so that calcification sites exist in the biological valve, which is an important reason for causing calcification of the biological valve. Meanwhile, due to wet film loading, the compressed size of the valve cannot be further reduced, the application range of a patient is reduced, the valve implantation path is limited, and vascular complications are increased.
With the gradual accumulation of TAVR clinical experience, complications are gradually reduced, and the indications tend to be gradually expanded to younger patients and patients with low or medium risk. Potential problems are gradually highlighted in the application process of all commercial transcatheter aortic valves at present, and mainly include the following:
1. the valve has insufficient durability and cannot meet the trend of valve patients to be younger;
2. the compressed size of the valve can not be further reduced, the adaptation range of a patient is reduced, the implantation path of the valve is limited, and vascular complications are increased;
3. the valve needs to be loaded before operation, is easy to calcify and inactivate, has poor durability, harsh manufacturing, storing and transporting conditions and the like, and limits the clinical application and popularization;
4. the biocompatibility of the existing artificial biological valve is still insufficient; the biocompatibility of the artificial biological valve is an important reason influencing the service life of the valve, and the elimination of the immunogenicity of the valve can enhance the creeping function of endothelial cells of a host and reduce the occurrence probability of perivalvular leakage.
Accordingly, the prior art is yet to be improved and developed.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a cross-linked artificial biological valve and a preparation method thereof, and aims to solve the problems that the existing artificial biological valve is insufficient in durability, the compressed size of the valve cannot be further reduced, the valve needs to be loaded before operation, is easy to calcify and inactivate, is harsh in manufacturing, storing and transporting conditions and is poor in biocompatibility.
The technical scheme of the invention is as follows:
a method of making a crosslinked bioprosthetic valve, comprising the steps of: immersing the artificial biological valve into a mixed solution consisting of glutaraldehyde and a polymer containing a specific chemical functional group, and carrying out cross-linking fixation to obtain a cross-linked artificial biological valve; wherein the specific chemical functional group is a hydroxyl group, a carboxyl group, an isocyanate group or a siloxane group.
The preparation method of the cross-linked artificial biological valve comprises the step of preparing the cross-linked artificial biological valve, wherein the polymer containing the specific chemical functional group comprises at least one of polyethylene glycol, polyvinyl alcohol, polyether dihydric alcohol, chitin, polyglycolic acid, polylactic acid, polyurethane and polysiloxane.
The preparation method of the cross-linked artificial biological valve comprises the step of preparing the cross-linked artificial biological valve, wherein the concentration of the glutaraldehyde is 0.1-5%.
The preparation method of the cross-linked artificial biological valve comprises the step of preparing the cross-linked artificial biological valve, wherein the concentration of the polymer containing the specific chemical functional group is 10-50%.
The preparation method of the cross-linked artificial biological valve is characterized in that the temperature for cross-linking and fixing is 25-45 ℃.
The preparation method of the cross-linked artificial biological valve comprises the step of cross-linking and fixing for 1-7 days.
The preparation method of the cross-linked artificial biological valve further comprises the following steps: and immersing the artificial biological valve into a mixed solution consisting of glutaraldehyde and the polymer containing the specific chemical functional group, carrying out cross-linking fixation, and taking out and completely drying at the temperature of 5-45 ℃.
A cross-linked bioprosthetic valve made by the method of any of the above.
Has the advantages that: the preparation method of the cross-linked artificial biological valve can enhance the physical and chemical properties and biocompatibility of the valve, prolong the service life of the valve, and simultaneously enable the valve to still keep flexible property in a dry state, thereby reducing the production and transportation costs of the valve and simplifying the use process of the valve.
Drawings
FIG. 1 is a schematic diagram of the equation for the chemical reaction of the valve of the present invention with a crosslinking agent and the polymer containing specific chemical functional groups (wherein the R group on the polymer represents a hydroxyl group, a carboxyl group, an isocyanate group or a siloxane group).
Detailed Description
The invention provides a cross-linked artificial biological valve and a preparation method thereof, and the invention is further described in detail below in order to make the purpose, technical scheme and effect of the invention clearer and more clear. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The invention creatively crosslinks the artificial biological valve (valve for short) by glutaraldehyde and a polymer mixed solution containing specific chemical functional groups, the chemical functional groups contained in the mixed solution react with groups in the valve chemically, and the valve can generate stable chemical covalent bonds with specific chemical functional groups such as hydroxyl, carboxyl, isocyanate or siloxane groups in the polymer while crosslinking with the glutaraldehyde, so that the flexibility and the compliance of the valve are improved, the valve can still maintain good physicochemical properties in a dry state, and the problem of material limitation of a pre-installed valve is solved.
It should be noted that the bioprosthetic valve of the present invention includes, but is not limited to, animal pericardial valves such as porcine pericardial valve and bovine pericardial valve.
The preparation method of the cross-linked artificial biological valve of the preferred embodiment of the invention comprises the following steps: immersing the artificial biological valve into a mixed solution consisting of glutaraldehyde and a polymer containing a specific chemical functional group, and carrying out cross-linking fixation to obtain a cross-linked artificial biological valve; wherein the specific chemical functional group is a hydroxyl group, a carboxyl group, an isocyanate group or a siloxane group.
Preferably, in a preferred embodiment of the present invention, the preparation method may further include: and immersing the artificial biological valve into a mixed solution consisting of glutaraldehyde and the polymer containing the specific chemical functional group, carrying out cross-linking fixation, and taking out and completely drying at the temperature of 5-45 ℃.
That is, in this embodiment, the method for preparing the cross-linked bioprosthetic valve may include two steps: 1. cross-linking of the biological valve; 2. and (5) drying at low temperature. The valve treated by the mixed solution is taken out and dried at a low temperature, and the obtained completely dried cross-linked artificial biological valve still has good flexibility and mechanical property.
The mixed solution composed of the glutaraldehyde and the polymer containing the specific chemical functional group is prepared by blending the glutaraldehyde and the polymer containing the specific chemical functional group to form a novel cross-linking agent formula, and a valve cross-linked by the formula can be dried and still soft and moist after being dried; conventional cross-linked valves, however, become brittle after drying, rendering them unusable.
When the specific chemical functional group is hydroxyl, taking a pig heart envelope valve as an example, the non-crosslinked pig heart envelope is collagen and contains a large amount of amino and carboxyl, and the amino reacts with aldehyde group and the carboxyl reacts with the hydroxyl to form covalent bonds through crosslinking of glutaraldehyde and a polymer blending solution containing the specific chemical functional group, so that the formed valve has better flexibility and can still maintain the dried state.
Further, in this embodiment, the polymer having a specific chemical functional group includes at least one of polyethylene glycol, polyvinyl alcohol, polyether glycol, chitin, polyglycolic acid, polylactic acid, polyurethane, and polysiloxane. The polymer solution containing the specific chemical functional group is a liquid which is nontoxic and harmless to a human body, and is an excellent toughening agent.
In this embodiment, when the specific chemical functional group is a hydroxyl group, the polymer containing the specific chemical functional group is polyethylene glycol, polyvinyl alcohol, polyether glycol or chitin; when the specific chemical functional group is a carboxyl group, the polymer containing the specific chemical functional group is polyglycolic acid or polylactic acid; when the specific chemical functional group is an isocyanate group, the polymer containing the specific chemical functional group is polyurethane; when the specific chemical functional group is a siloxane group, the polymer containing the specific chemical functional group is a polysiloxane.
The invention adopts the polymer with the toughening effect and containing the specific chemical functional group, and can be combined with the surface of the valve through a chemical covalent bond, so that the valve also has good flexibility, and the valve can still keep excellent flexibility under a dry condition, thereby the crosslinking method can be applied to the preparation of the dry valve, and the dry storage of the valve is realized.
The cross-linking fixing method combines the polymer containing the specific chemical functional group with the soft and smooth function with the valve through chemical covalent bond, so that the polymer covers the surface of the valve, the flexibility of the polymer is endowed to the valve, and meanwhile, one part of the polymer containing the specific chemical functional group is connected to the inside of the tissue of the valve, thereby further enhancing the toughening effect.
The invention adopts artificial biological valve to crosslink by mixed solution of glutaraldehyde and hydroxyl-containing polymer. The glutaraldehyde and the polymer containing specific chemical functional groups in the solution are combined with the surface of the valve through chemical covalent bonds, so that the valve has good physical and chemical properties and good flexibility, and can still keep good flexibility under a dry condition, and the dry storage of the valve is realized. Meanwhile, the method reduces the thickness of the valve material, has the function of fold self-flattening, deactivates tissues, reduces the immunogenicity of the material, enhances the creeping capacity of endothelial cells of a host, reduces the incidence rate of long-term perivalvular leakage and prolongs the service life of the biological valve.
As shown in FIG. 1, FIG. 1 shows the equation of the chemical reaction between the bioprosthetic valve of the present invention and the cross-linking agent and the polymer having the specific chemical functional group. In the invention, glutaraldehyde and amino in the valve are subjected to cross-linking reaction through aldehyde groups, so that the physical and chemical properties of the valve are enhanced; the polymer containing the specific chemical functional group chemically reacts with amino or hydroxyl in the valve through the specific chemical functional group at the tail end, and the surface of the valve is covered with a layer of polymer while the valve is crosslinked. The polymer with larger molecular weight can cover a larger range of the valve surface by a single polymer molecule, so the polymer containing the specific chemical functional group can easily endow the chemical and physical properties of the polymer to the artificial biological valve.
More specifically, as shown in fig. 1, glutaraldehyde undergoes a cross-linking reaction with an amino group in the valve via an aldehyde group; the polymer is subjected to chemical reaction with carboxyl in the valve through terminal hydroxyl, or the terminal carboxyl is subjected to chemical reaction with amino or hydroxyl in the valve, or terminal isocyanate is subjected to reaction with the amino or hydroxyl in the valve, or terminal siloxane is subjected to reaction with the amino in the valve, and the surface of the valve is covered with a layer of polymer while the cross-linking effect is achieved.
Further, in this embodiment, the concentration of the glutaraldehyde is 0.1% to 5%. For example, the concentration of glutaraldehyde may be 0.1%, 1%, 2%, 3%, 4%, or 5%.
Further, in this embodiment, the concentration of the polymer having the specific chemical functional group is 10% to 50%. For example, the concentration of the polymer containing a particular chemical functional group may be 10%, 20%, 30%, 40%, or 50%.
Further, in this embodiment, the temperature for cross-linking and fixing is 25-45 ℃. For example, the temperature for crosslinking fixation may be 25 ℃, 30 ℃, 35 ℃, 40 ℃ or 45 ℃.
Furthermore, in this embodiment, the time for the cross-linking and fixing is 1 to 7 days. For example, the time for cross-linking fixation may be 1 day, 2 days, 3 days, 4 days, or 5 days.
In specific implementation, the step of cross-linking the bioprosthetic valve of the present invention may be: immersing the artificial biological valve into a mixed solution of glutaraldehyde and a polymer containing a specific chemical functional group for crosslinking, wherein the concentration of the glutaraldehyde is 0.1-5%, the concentration of the polymer containing the specific chemical functional group is 10-50%, the crosslinking time is 1-7 days, and meanwhile, the temperature is controlled to be 25-45 ℃, and the valve is ensured to be flatly unfolded.
The invention also provides a cross-linked artificial biological valve which is prepared by the preparation method of the cross-linked artificial biological valve.
By adopting the preparation method of the cross-linked artificial biological valve, the valve leaf is thinner, firmer, more durable, better in biocompatibility and more stable in property. The reduction of the thickness of the valve leaflet reduces the overall dimension of the valve after being compressed by 80%, and the method can be applied to the preparation of a dry valve, so that the valve can be preassembled in a delivery system and delivered out of a factory, can be used at any time, does not need preoperative installation, shortens the operation time, and reduces the death risk caused during the period of waiting for the implantation of the valve in the operation of a patient. The valve cross-linked by the method does not need to be placed in a presettable system stored in a storage solution containing glutaraldehyde, so that the problem of valve durability is thoroughly solved, the biocompatibility of the valve is improved, the endocarditis occurrence probability caused by immunogen risks is reduced, the host endothelial cell climbing function is enhanced, and the perivalvular leakage occurrence probability is reduced.
The invention is illustrated in detail below with specific examples:
example 1
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 5% of glutaraldehyde, 10% of polyethylene glycol and 20% of polyvinyl alcohol at 45 ℃ for 2 days, taking out and completely drying at 45 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 2
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 4% of glutaraldehyde and 20% of polyether glycol for 5 days at the temperature of 40 ℃, taking out the porcine pericardium valve and completely drying the porcine pericardium valve at the temperature of 35 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 3
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 3% glutaraldehyde and 30% chitin for 3 days at 35 ℃, taking out and completely drying at 35 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 4
Cross-linking and soaking the porcine pericardium valve in a mixed solution of 2% of glutaraldehyde, 10% of polyglycolic acid and 20% of polylactic acid for 1 day at the temperature of 30 ℃, taking out and completely drying at the temperature of 25 ℃ to obtain a cross-linked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 5
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 1% glutaraldehyde and 40% polyurethane at 35 ℃ for 1 day, taking out and completely drying at 25 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 6
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 0.1% glutaraldehyde and 50% polysiloxane at 25 ℃ for 1 day, taking out and completely drying at 5 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 7
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 0.1 percent of glutaraldehyde and 10 percent of polysiloxane at the temperature of 25 ℃ for 1 day, taking out and completely drying at the temperature of 5 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
Example 8
Crosslinking and soaking the porcine pericardium valve in a mixed solution of 0.1 percent of glutaraldehyde and 10 percent of polyethylene glycol for 7 days at the temperature of 25 ℃, taking out and completely drying at the temperature of 5 ℃ to obtain a crosslinked artificial biological valve; the detection shows that the cross-linked artificial biological valve prepared by the embodiment still keeps excellent flexibility after being dried, the external dimension of the compressed valve is reduced by 80%, and the cross-linked artificial biological valve has good biocompatibility and stable properties.
In summary, the present invention provides a bioprosthetic valve and a preparation method thereof, wherein the preparation method comprises the steps of: immersing the artificial biological valve into a mixed solution consisting of glutaraldehyde and a polymer containing a specific chemical functional group, and carrying out cross-linking fixation to obtain a cross-linked artificial biological valve; wherein the specific chemical functional group is a hydroxyl group, a carboxyl group, an isocyanate group or a siloxane group. The preparation method of the cross-linked artificial biological valve can enhance the physical and chemical properties and biocompatibility of the valve, prolong the service life of the valve, and simultaneously enable the valve to still keep flexible property in a dry state, thereby reducing the production and transportation costs of the valve and simplifying the use process of the valve.
It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.