CN114652893A - Method for treating bioprosthetic tissue and bioprosthetic heart valve - Google Patents
Method for treating bioprosthetic tissue and bioprosthetic heart valve Download PDFInfo
- Publication number
- CN114652893A CN114652893A CN202011573018.2A CN202011573018A CN114652893A CN 114652893 A CN114652893 A CN 114652893A CN 202011573018 A CN202011573018 A CN 202011573018A CN 114652893 A CN114652893 A CN 114652893A
- Authority
- CN
- China
- Prior art keywords
- solution
- bioprosthetic tissue
- tissue
- capping
- bioprosthetic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/20—Materials or treatment for tissue regeneration for reconstruction of the heart, e.g. heart valves
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Transplantation (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Urology & Nephrology (AREA)
- Vascular Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Materials For Medical Uses (AREA)
- Prostheses (AREA)
Abstract
According to the method for treating the bioprosthetic tissue and the bioprosthetic heart valve, provided by the invention, the calcification degree of the bioprosthetic tissue can play a relatively obvious relieving effect after the end capping treatment and the reduction treatment are respectively carried out on the at least partially crosslinked bioprosthetic tissue in two steps, and the mechanical performance of the bioprosthetic tissue is not influenced.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a method for treating bioprosthetic tissue and a bioprosthetic heart valve.
Background
At present, biological valves and biological patches commonly used in clinic, such as animal-derived collagen fiber tissue materials like bovine pericardium and small intestine mucosa, are usually subjected to cross-linking and fixing treatment by using chemical reagents such as glutaraldehyde and/or formaldehyde storage. Such fixation methods often result in residual aldehyde groups present in the bioprosthetic tissue, and numerous studies have shown that formaldehyde/glutaraldehyde-fixed bioprosthetic tissue causes in vivo calcification after implantation.
Methods for anti-calcification of bioprosthetic tissue have been reported as follows: decellularization treatment, alcohol treatment, surfactant treatment, metal ion competition, and the like. Among them, the decellularization process can remove most of antigens and DNA, but biological tissues become loose, and thus the mechanical properties thereof are degraded; alcohol treatment, which has good short-term anti-calcification effect, but has high long-term calcification risk due to the existence of residual aldehyde group; the surfactant treatment can remove phospholipids in bioprosthetic tissues, but aldehyde groups still exist as calcification-inducing factors; the metal ion competition can be combined with the existing combination sites of the bioprosthetic tissue to play a certain anti-calcification effect, but the induction effect of the aldehyde group long-term calcification cannot be completely solved. That is, although the above method has a certain anti-calcification effect, the anti-calcification effect is not good in practical use, and there is a risk of a decrease in mechanical properties.
Disclosure of Invention
The invention aims to solve the problems that the prior art bioprosthetic tissue treatment method and bioprosthetic heart valve have poor anti-calcification effect and are accompanied by the risk of reduced mechanical performance.
In order to solve the above problems, the present invention provides a method for treating bioprosthetic tissue, comprising:
step S10: providing bioprosthetic tissue, wherein the bioprosthetic tissue is at least partially cross-linked;
step S20: performing end capping treatment on the bioprosthetic tissue by using an end capping agent;
step S30: and (3) carrying out reduction treatment on the bioprosthetic tissue subjected to the end capping treatment by using a reducing agent.
Optionally, the step S30 includes: placing the bioprosthetic tissue subjected to the end capping treatment in a reducing solution consisting of the reducing agent and a phosphate buffer solution, and subjecting the bioprosthetic tissue subjected to the end capping treatment to a reducing treatment by the reducing agent under first predetermined conditions.
Optionally, the pH value of the reducing solution is 6.8-8.6, the concentration of the reducing agent in the reducing solution is 0.02-2.0% W/V, and the concentration of the phosphate buffer solution in the reducing solution is greater than 0.05M.
Optionally, the first predetermined condition includes: the method comprises the steps of obtaining a first preset temperature, a first preset rotating speed and a first preset oscillation time, wherein the first preset temperature is 15-50 ℃, the first preset rotating speed is 50-200 rpm, and the first preset oscillation time is 2-96 hours.
Optionally, after subjecting the reducing agent to a reduction treatment on the bioprosthetic component, the method further comprises: and replacing the reduction solution after the reduction treatment at least once by using the reduction solution, and enabling the reducing agent in the replaced reduction solution to continue the reduction treatment on the bioprosthetic tissue under a first preset condition.
Optionally, the number of times of replacing the reducing solution is 1-5 times.
Optionally, the reducing agent is a hydride or a sulfide.
Optionally, the reducing agent is at least one selected from sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, diisobutylaluminum hydride, diisobutyl potassium hydride, sodium bisulfite, and sodium thiosulfate.
Optionally, the method for performing the end capping treatment on the bioprosthetic tissue comprises: placing the bioprosthetic tissue in a capping solution consisting of a capping agent and a phosphate buffer solution, and allowing the capping solution to treat the bioprosthetic tissue under second predetermined conditions.
Optionally, the pH value of the end-capping solution is 6.8 to 8.6, the concentration of the end-capping agent in the end-capping solution is 0.02% to 2.0% W/V, and the concentration of the phosphate buffer solution in the end-capping solution is greater than 0.05M.
Optionally, the second predetermined condition includes: the temperature control device comprises a second preset temperature, a second preset rotating speed and a second preset oscillation time, wherein the second preset temperature is 15-50 ℃, the second preset rotating speed is 50-200 rpm, and the second oscillation time is 2-96 hours.
Optionally, the end-capping agent is selected from at least one of amino acids, saturated alkylamines, unsaturated alkylamines, aminoalcohols, polyetheramines, aminoalkylacids, aminoalkyldiacids, amino surfactants.
Optionally, after the end capping treatment, the method further includes: and cleaning the bioprosthesis tissue for the second time by using a second cleaning solution, wherein the second cleaning solution is one selected from an ethanol solution, normal saline or a phosphate buffer solution, the number of times of cleaning for the second time is 3-5, and each time of cleaning is 3-5 min.
In order to solve the above problems, the present invention also provides a bioprosthetic heart valve comprising bioprosthetic tissue treated according to the method for treating a bioprosthetic assembly described in any one of the above.
In the invention, after the end capping treatment and the reduction treatment are respectively carried out on the at least partially crosslinked bioprosthetic tissue in two steps, a relatively obvious effect of reducing the calcification degree of the bioprosthetic tissue can be achieved, and the mechanical property of the bioprosthetic tissue is not influenced.
Drawings
FIG. 1 is a schematic flow diagram of a method of treating bioprosthetic tissue of the present invention.
FIG. 2 is a comparison graph of calcium content of bovine pericardial tissue treated under different experimental conditions and implanted in rats for 8 weeks.
Detailed Description
The bioprosthetic tissue and the bioprosthetic heart valve treatment method according to the present invention will be described in further detail with reference to the accompanying drawings and specific examples. The advantages and features of the present invention will become more apparent from the following description. It is to be noted that the drawings are in a very simplified form and are not to precise scale, which is merely for the purpose of facilitating and distinctly claiming the embodiments of the present invention.
FIG. 1 is a schematic flow diagram of a method of treating bioprosthetic tissue of the present invention. FIG. 2 is a comparison graph of calcium content of bovine pericardial tissue treated under different experimental conditions and implanted in rats for 8 weeks. A method of reducing calcification in bioprosthetic tissue in accordance with the present invention is described in detail below with reference to FIGS. 1 and 2.
As shown in fig. 1, the present embodiment provides a method for treating bioprosthetic tissue, including:
step S10 is executed: providing bioprosthetic tissue, wherein the bioprosthetic tissue is at least partially cross-linked.
Wherein, in this embodiment, the groups generated by the bioprosthetic tissue during the cross-linking process are associated with toxicity, calcification, and immunogenicity within the bioprosthetic tissue. Specifically, the group is at least one of an aldehyde group and a carboxylic acid group.
And, in this embodiment, the bioprosthetic tissue includes, but is not limited to, tissue selected from the group consisting of pericardium, heart valves, tectorial membranes, pleura, small intestine submucosa, dura mater, ligament, tendon, blood vessel, intravesical membrane, or skin, wherein the pericardium may be selected from the group consisting of bovine pericardium or porcine pericardium, preferably bovine pericardium.
Also, in this embodiment, the bioprosthetic tissue may be crosslinked using glutaraldehyde or formaldehyde. For example, the crosslinking treatment may be carried out using a 0.625% W/V glutaraldehyde solution.
In this embodiment, after providing the bioprosthetic tissue, the method further comprises: and cleaning the bioprosthesis tissue for the first time by using a first cleaning solution, wherein the first cleaning solution is selected from one of normal saline, phosphate buffer solution or ethanol solution, the number of times of the first cleaning is 3-5, and each cleaning is carried out for 3-5 min. Therefore, glutaraldehyde or formaldehyde which does not participate in the reaction is sufficiently washed away, so that the treatment effect is prevented from being interfered when the bioprosthetic tissue is subsequently treated.
In the present embodiment, if the first washing is performed using the phosphate buffer solution, the pH of the phosphate buffer solution used for the first washing is 6.8 to 8.6, and the concentration is 0.05M to 0.2M. And if the first washing is performed by using an ethanol solution, the concentration of the ethanol solution used for the first washing is 20-80%.
Step S20 is performed to perform a capping treatment on the bioprosthetic tissue using a capping solution.
In this embodiment, "end-capping" refers to blocking, removing, or altering functional groups that would have an adverse effect on the tissue properties of the bioprosthesis. For example, ethanolamine is added to cap incompletely reacted aldehyde groups on the bioprosthetic tissue. This "capping" may also be referred to as capping, grafting, and the like.
Specifically, in this embodiment, the capping agent is selected from at least one of amino acids, saturated alkylamines, unsaturated alkylamines, aminoalcohols, polyether amines, aminoalkylacids, aminoalkyldiacids, and amino surfactants. Preferably, it is selected from amino alcohols, 2-aminoalkylacids, aminoalkyldiacids. More preferably, the capping agent is selected from ethanolamine, 2-aminooleic acid.
And, the method of capping the bioprosthetic tissue comprises: placing the bioprosthetic tissue in a capping solution consisting of the capping agent and a phosphate buffer solution to cause the capping agent to treat the bioprosthetic tissue under second predetermined conditions. The pH value of the end capping solution is 6.8-8.6, the concentration of the end capping agent in the end capping solution is 0.02-2.0% W/V, the concentration of the phosphate buffer solution in the end capping solution is more than 0.05M, and preferably, the concentration of the phosphate buffer solution in the end capping solution is 0.05-0.2M.
Further, in this embodiment, the second predetermined condition includes: the temperature control device comprises a second preset temperature, a second preset rotating speed and a second preset oscillation time, wherein the second preset temperature is 15-50 ℃, the second preset rotating speed is 50-200 rpm, and the second oscillation time is 2-96 hours. In this manner, the bioprosthetic tissue is adequately capped.
Further, in this embodiment, after the end-capping treatment is performed, the method further includes: and cleaning the bioprosthesis tissue for the second time by using a second cleaning solution, wherein the second cleaning solution is one selected from an ethanol solution, normal saline or a phosphate buffer solution, the number of times of cleaning for the second time is 3-5, and each time of cleaning is 3-5 min. Thus, the free blocking agent can be sufficiently removed, so as to reduce the influence of the blocking agent on the subsequent processing steps. And when the second cleaning solution is an ethanol solution, the concentration of the ethanol solution for the second cleaning is 10-50% V/V. When the second cleaning solution is a phosphate buffer solution, the pH value of the phosphate buffer solution for the second cleaning is 6.8-8.6, and the concentration of the phosphate buffer solution is 0.05-0.2M.
Step S30: and carrying out reduction treatment on the bioprosthetic tissue subjected to the end capping treatment by using a reduction solution.
In this embodiment, the treatment of the bioprosthetic tissue with the reducing solution means that the capped bioprosthetic component is reduced by reacting the reducing agent with the capped bioprosthetic component in step S20. In the step, the reducing agent mainly reacts with the Schiff base generated in the cross-linking and end-capping step to reduce the Schiff base, thereby stabilizing chemical bonds in the bioprosthetic tissue.
In this embodiment, the reducing agent is a hydride or a sulfide, wherein the hydride is preferably a borohydride selected from at least one of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, diisobutyl aluminum hydride, and diisobutyl potassium hydride. Wherein, when the reducing agent is sodium cyanoborohydride and sodium triacetoxyborohydride, the reduction effect is better. Further, the sulfide may include sodium bisulfite, sodium thiosulfate, or the like.
And, in this embodiment, the step S30 includes: placing the bioprosthetic tissue in a reducing solution consisting of a reducing agent and a phosphate buffer solution to cause the reducing agent to perform a reducing treatment on the bioprosthetic tissue after the capping treatment under first predetermined conditions.
Wherein the pH value of the reducing solution is 6.8-8.6, the concentration of the reducing agent in the reducing solution is 0.02-2.0% W/V, and more preferably, the concentration of the reducing agent can be 0.05-0.5% W/V. The concentration of the phosphate buffer solution in the reducing solution is more than 0.05M, and preferably, the concentration of the phosphate buffer in the reducing solution is 0.05M to 0.2M.
Further, in the present embodiment, the first predetermined condition includes: the device comprises a first preset temperature, a first preset rotating speed and a first preset oscillation time, wherein the first preset temperature is 15-50 ℃, the first preset rotating speed is 50-200 rpm, and the first preset oscillation time is 2-96 hours.
Further, after treating the bioprosthetic tissue with a reducing agent, the method further comprises: storing, drying or sterilizing the bioprosthetic tissue treated with the reducing agent.
Wherein, the bioprosthetic tissue treated by the reducing agent can be placed in a liquid for liquid storage, and optionally, the liquid can be a glutaraldehyde storage solution. Placing the bioprosthetic tissue in the liquid for storage may also sterilize the bioprosthetic tissue in addition to maintaining the properties of the bioprosthetic tissue.
The method of drying treatment may include: sequentially immersing the bioprosthetic tissue into a carbohydrate solution with increased solubility gradient at room temperature for dehydration; the saccharide solution is a water-absorbing and moisture-retaining monosaccharide, disaccharide, trisaccharide, polysaccharide or sugar alcohol saccharide solution; preferably, the saccharide solution is fructose, sucrose, trehalose, amorphous raffinose, chitosan and chitosan-modified polysaccharide, sorbitol or mannitol solution. In some embodiments of the invention, the bioprosthetic tissue is sequentially immersed in aqueous solutions of sucrose having a solubility of 30%, 40%, 50%, 55%, 60%, 65% (w/v), respectively, for 30min at room temperature, and the pericardium is then removed and dried on a fibrous desiccant. And the sterilization treatment may be EO (ethylene oxide) sterilization.
In this embodiment, the subsequent processing manner of the bioprosthetic tissue after the capping treatment and the reducing treatment is not specifically limited herein, and the actual situation is the standard.
Experiments one to six are performed below to treat the bioprosthetic tissue, which is bovine pericardial tissue, and further illustrate the beneficial effects of the method for treating the bioprosthetic tissue according to the present embodiment. In the first experiment, the bioprosthetic tissue is not subjected to end capping and reduction treatment; in the second experiment, only the bioprosthetic tissue is subjected to end capping treatment and is not subjected to reduction treatment; in the third experiment, the end capping treatment and the reduction treatment are synchronously carried out on the bioprosthetic tissue in the same step; in experiment four, experiment five and experiment six, the end capping treatment and the reduction treatment in the bioprosthetic tissue are performed step by step, wherein the reduction conditions of experiment four, experiment five and experiment six are different. In the above experiment, the bovine pericardial tissue is subjected to the cross-linking treatment under the same conditions before the experiment, that is, the bovine pericardial tissue before the cross-linking treatment is placed in a 0.625% glutaraldehyde solution for the cross-linking treatment.
Experiment one:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
Then, the bovine pericardium is dehydrated and dried, namely, the bovine pericardium is sequentially immersed into sucrose aqueous solutions with the solubility of 30%, 40%, 50%, 55%, 60% and 65% (w/v) respectively at room temperature for 30min each time, and then the bovine pericardium is taken out and dried on a fiber drying agent.
Experiment two:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
And then, carrying out end-capping treatment, namely preparing 1L of end-capping solution, putting the cleaned bovine pericardium tissue into the end-capping solution, placing the bovine pericardium tissue on a constant-temperature shaking table, oscillating the bovine pericardium tissue at a constant temperature of 25 ℃ for 24 hours at a rotating speed of 60rpm, and cleaning the bovine pericardium tissue for 5 times by using 0.1M sterile phosphate buffer solution for 5min each time. Wherein the blocking solution is a phosphate buffer solution of 8mM ethanolamine, and the concentration of the phosphate buffer solution is 0.1M. The pH value of the end capping solution is between 7.40 +/-0.1.
And finally, performing dehydration drying treatment, namely sequentially immersing the bovine pericardium into sucrose aqueous solutions with the solubilities of 30%, 40%, 50%, 55%, 60% and 65% (w/v) at room temperature for 30min each time, and taking out the bovine pericardium and drying on a fiber drying agent.
Experiment three:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
And then, carrying out end-capping reduction treatment, namely preparing 1L of end-capping reduction solution, putting the cleaned bovine pericardium tissue into the end-capping reduction solution, placing the solution on a constant-temperature shaking table, oscillating the solution at a constant temperature of 25 ℃ for 4 hours at a rotating speed of 60rpm, cleaning the solution for 5 times by using 0.1M sterile phosphate buffer solution after the end of the process, cleaning the solution for 5 minutes each time, and cleaning the solution for 5 times by using 20% ethanol, and cleaning the solution for 5 minutes each time. Wherein the end-capping reduction solution is a phosphate buffer solution of 8mM ethanolamine and 0.4% sodium borohydride (W/V), and the concentration of the phosphate buffer solution is 0.1M.
And finally, performing dehydration drying treatment, namely sequentially immersing the bovine pericardium into sucrose aqueous solutions with the solubilities of 30%, 40%, 50%, 55%, 60% and 65% (w/v) at room temperature for 30min each time, and taking out the bovine pericardium and drying on a fiber drying agent.
Experiment four:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
And then, carrying out end-capping treatment, namely preparing 1L of end-capping solution, putting the cleaned bovine pericardium tissue into the end-capping solution, placing the bovine pericardium tissue on a constant-temperature shaking table, oscillating the bovine pericardium tissue at a constant temperature of 25 ℃ for 24 hours at a rotating speed of 60rpm, and cleaning the bovine pericardium tissue for 5 times by using 0.1M sterile phosphate buffer solution for 5min each time. Wherein the end capping solution is phosphate buffer solution of 8mM ethanolamine, the concentration of the phosphate buffer solution is 0.1M, and the pH value of the end capping solution is 7.40 +/-0.1.
And then, carrying out reduction treatment, namely, assembling the capped bovine pericardium into a reduction solution, oscillating at a constant temperature of 25 ℃ for 4 hours at a rotation speed of 60rpm, and then washing for 5 times by using 20% ethanol for 5min each time. Wherein the reducing solution is 0.4% sodium triacetoxyborohydride (W/V) phosphate buffer solution, and the concentration of the phosphate buffer solution is 0.1M.
And finally, performing dehydration drying treatment, namely sequentially immersing the bovine pericardium into sucrose aqueous solutions with the solubilities of 30%, 40%, 50%, 55%, 60% and 65% (w/v) at room temperature for 30min each time, and taking out the bovine pericardium and drying on a fiber drying agent.
Experiment five:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
And then, carrying out end-capping treatment, namely preparing 1L of end-capping solution, putting the cleaned bovine pericardium tissue into the end-capping solution, placing the bovine pericardium tissue on a constant-temperature shaking table, oscillating the bovine pericardium tissue at a constant temperature of 25 ℃ for 24 hours at a rotating speed of 60rpm, and cleaning the bovine pericardium tissue for 5 times by using 0.1M sterile phosphate buffer solution for 5min each time. Wherein the end capping solution is phosphate buffer solution of 8mM ethanolamine, the concentration of the phosphate buffer solution is 0.1M, and the pH value of the end capping solution is 7.40 +/-0.1.
And then, carrying out reduction treatment, namely, assembling the capped bovine pericardium into a reduction solution, and oscillating for 2 hours at a constant temperature of 25 ℃ at a rotation speed of 60 rpm. Then, the solution was washed 5 times with 20% ethanol for 5min each time. Wherein the reducing solution is a phosphate buffer solution of 0.05% sodium borohydride (W/V), and the concentration of the phosphate buffer solution is 0.1M.
And finally, performing dehydration drying treatment, namely sequentially immersing the bovine pericardium into sucrose aqueous solutions with the solubilities of 30%, 40%, 50%, 55%, 60% and 65% (w/v) at room temperature for 30min each time, and taking out the bovine pericardium and drying on a fiber drying agent.
Experiment six:
first, the bovine pericardial tissue was washed 3 times in 20% ethanol for 5min each.
And then, carrying out end-capping treatment, namely preparing 1L of end-capping solution, putting the cleaned bovine pericardium tissue into the end-capping solution, placing the bovine pericardium tissue on a constant-temperature shaking table, oscillating the bovine pericardium tissue at a constant temperature of 25 ℃ for 24 hours at a rotating speed of 60rpm, and cleaning the bovine pericardium tissue for 5 times by using 0.1M sterile phosphate buffer solution for 5min each time. Wherein the blocking solution is a phosphate buffer solution of 8mM ethanolamine, the concentration of the phosphate buffer solution is 0.1M, and the pH value of the blocking solution is 7.40 +/-0.1.
And then, carrying out reduction treatment, namely, assembling the capped bovine pericardium into a reduction solution, oscillating at a constant temperature of 25 ℃ for 2h at a rotation speed of 60rpm, and then washing for 5 times by using 20% ethanol for 5min each time. Wherein the reducing solution is a phosphate buffer solution of 0.4% sodium cyanoborohydride (W/V), and the concentration of the phosphate buffer solution is 0.1M.
And finally, performing dehydration drying treatment, namely sequentially immersing the bovine pericardium into sucrose aqueous solutions with the solubilities of 30%, 40%, 50%, 55%, 60% and 65% (w/v) at room temperature for 30min each time, and taking out the bovine pericardium and drying on a fiber drying agent.
In addition, in the first to sixth experiments, the last step does not affect the experimental effect, and in this embodiment, the last step is drying treatment, and optionally, liquid storage treatment may be performed.
And after the experiment is finished, carrying out mechanical tensile test in a solution environment after the dehydrated and dried bovine pericardial tissue is rehydrated. Wherein, the length of the bovine pericardium tissue in each experiment is 50mm, the width is 5mm, the length of the test gauge is set to be 25mm, the tensile rate is 100mm/min on a universal material testing machine, and the tensile test is carried out until the test sample is broken. Then, acquiring the breaking strength and the breaking elongation of the test sample, wherein the breaking strength refers to the ratio of the tensile force when the test sample breaks to the breaking cross-sectional area when the mechanical tensile test is carried out; elongation at break refers to the ratio of the length of the test specimen after stretching to the length before stretching when tested in mechanical tensile. Wherein, the higher the breaking strength, the better the mechanical property of the material, and the larger the breaking elongation, the better the elasticity of the material.
The mechanical tensile results of the bovine pericardium tissue after the six experimental treatments are shown in the table below.
| Item | Experiment one | Experiment two | Experiment three | Experiment four | Experiment five | Experiment six |
| Breaking strength/Mpa | 11.89±5.61 | 12.59±6.34 | 8.01±4.31 | 13.24±6.41 | 12.21±6.36 | 12.94±7.83 |
| Elongation at break/% | 28.6±5.9 | 26.8±6.2 | 36.6±6.6 | 28.0±5.7 | 27.3±7.7 | 28.6±9.56 |
Watch 1
From the above results, it can be seen that the rupture strength in the mechanical property test of the bovine pericardial tissue treated in the first experiment, the second experiment, the fourth experiment, the fifth experiment and the sixth experiment was substantially the same. However, when the capping treatment and the reducing treatment are performed simultaneously in the same step, the rupture strength of the treated bovine pericardium tissue in experiment three is obviously lower than that of the treated bovine pericardium tissue in experiment one, experiment two, experiment four, experiment five and experiment six. That is, the mechanical properties of the bioprosthetic tissue are poor when the capping treatment and the reducing treatment are performed simultaneously in the same step.
In this example, bovine pericardial tissue treated in the above experiment was implanted into rats of 3 weeks old from lactation, and the samples were taken out after 8 weeks of implantation and dried in a constant temperature oven at 80 ℃ for 48 hours to a constant weight. Then, calcium content of the bovine pericardium tissue treated in the above experiment was measured by a flame-atomic absorption spectrophotometer, and the test results are shown in fig. 2.
As can be seen from fig. 2, in the present experiment, the bovine pericardium tissue in the first experiment was not subjected to the capping treatment and the reduction treatment, and the degree of calcification was high. The bovine pericardial tissue in the second experiment is not subjected to reduction treatment, the calcification degree of the bovine pericardial tissue is lower than that of the bovine pericardial tissue which is not subjected to blocking treatment and reduction treatment in the first experiment, but is still higher than that of the bovine pericardial tissue subjected to blocking treatment and reduction treatment in the third experiment in the same step, and is also higher than that of the bovine pericardial tissue respectively treated in the two steps of blocking treatment and reduction treatment in the fourth experiment, the fifth experiment and the sixth experiment, and the bovine pericardial tissue subjected to reduction treatment in the third experiment, the fourth experiment, the fifth experiment and the sixth experiment shows good calcification reduction effect. Therefore, in the embodiment, after the bovine pericardium tissue is subjected to the step-by-step end-capping treatment and the calcification treatment, the calcification degree of the bovine pericardium tissue is greatly reduced, and the mechanical property of the bovine pericardium tissue is not affected.
In summary, in this embodiment, after the capping treatment and the reducing treatment are respectively performed in two steps on the at least partially crosslinked bioprosthetic tissue, a relatively significant reduction effect can be achieved on the calcification degree of the bioprosthetic tissue, and the mechanical properties of the bioprosthetic tissue are not affected.
Further, in this embodiment, after the reduction treatment of the bioprosthetic tissue with the reducing agent, the method further comprises: replacing the reduction solution after the bioprosthetic tissue is treated at least once by using the reduction solution without treating the bioprosthetic tissue, and enabling the reducing agent in the replaced reduction solution to continuously perform reduction treatment on the bioprosthetic tissue under a first preset condition. Wherein the number of times of replacing the reducing solution is 1-5 times. Therefore, the bioprosthesis component subjected to the end capping treatment is subjected to at least two times of reduction treatment, and the bioprosthesis subjected to the end capping treatment can be reduced more completely by the at least two times of reduction treatment, so that the reduction effect is further improved, and further, the chemical bond in the bioprosthesis tissue is more stable. Further, the amount of the reducing agent used per reduction treatment is smaller than that used in the case where only one reduction is performed. In this manner, the reducing environment for each reduction is made milder to reduce the impact of the reduction process on the bioprosthetic tissue.
Furthermore, in this embodiment, the reducing solution of the bioprosthetic tissue that has been treated may be replaced with the reducing solution of the bioprosthetic tissue that has not been treated in the same vessel. The bioprosthetic tissue that has been subjected to reduction treatment may also be removed from the reducing solution in which the bioprosthetic tissue has been treated, and then placed in another reducing solution with untreated bioprosthetic tissue for further treatment. The specific situation is not limited herein.
Further, the present embodiment also discloses a bioprosthetic heart valve comprising bioprosthetic tissue treated by the bioprosthetic component treatment method according to any one of the above items.
The above description is only for the purpose of describing the preferred embodiments of the present invention, and is not intended to limit the scope of the present invention, and any variations and modifications made by those skilled in the art based on the above disclosure are within the scope of the appended claims.
Claims (14)
1. A method of treating bioprosthetic tissue, comprising:
step S10: providing bioprosthetic tissue, wherein the bioprosthetic tissue is at least partially cross-linked;
step S20: performing end capping treatment on the bioprosthetic tissue by using an end capping agent;
step S30: and (3) carrying out reduction treatment on the bioprosthetic tissue subjected to the end capping treatment by using a reducing agent.
2. The method for treating bioprosthetic tissue of claim 1, wherein the step S30 comprises: placing the bioprosthetic tissue subjected to the end capping treatment in a reducing solution consisting of the reducing agent and a phosphate buffer solution, and subjecting the bioprosthetic tissue subjected to the end capping treatment to a reducing treatment by the reducing agent under first predetermined conditions.
3. The method of treating bioprosthetic tissue of claim 2, wherein the pH of the reducing solution is 6.8 to 8.6, the concentration of the reducing agent in the reducing solution is 0.02% to 2.0% W/V, and the concentration of the phosphate buffer solution in the reducing solution is greater than 0.05M.
4. The method of treating bioprosthetic tissue of claim 2, wherein the first predetermined condition comprises: the device comprises a first preset temperature, a first preset rotating speed and a first preset oscillation time, wherein the first preset temperature is 15-50 ℃, the first preset rotating speed is 50-200 rpm, and the first preset oscillation time is 2-96 hours.
5. The method of treating bioprosthetic tissue of claim 2, wherein after subjecting the reducing agent to a reducing treatment on the bioprosthetic component, the method further comprises: and replacing the reduction solution after the reduction treatment at least once by using the reduction solution, and enabling the reducing agent in the replaced reduction solution to continue the reduction treatment on the bioprosthetic tissue under a first preset condition.
6. The method of treating bioprosthetic tissue of claim 5, wherein the number of times the reducing solution is replaced is 1 to 5 times.
7. The method of treating bioprosthetic tissue of claim 1, wherein the reducing agent is a hydride or a sulfide.
8. The method of treating bioprosthetic tissue of claim 7, wherein the reducing agent is selected from at least one of sodium borohydride, potassium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, diisobutylaluminum hydride, diisobutyl potassium hydride, sodium bisulfite, and sodium thiosulfate.
9. The method of treating bioprosthetic tissue of claim 1, wherein the method of capping the bioprosthetic tissue comprises: placing the bioprosthetic tissue in a capping solution consisting of a capping agent and a phosphate buffer solution, and allowing the capping solution to treat the bioprosthetic tissue under second predetermined conditions.
10. The method of claim 9, wherein the capping solution has a pH of 6.8 to 8.6, the capping agent concentration in the capping solution is 0.02% to 2.0% W/V, and the phosphate buffer solution concentration in the capping solution is greater than 0.05M.
11. The method of treating bioprosthetic tissue of claim 9, wherein the second predetermined condition comprises: the temperature control device comprises a second preset temperature, a second preset rotating speed and a second preset oscillation time, wherein the second preset temperature is 15-50 ℃, the second preset rotating speed is 50-200 rpm, and the second oscillation time is 2-96 hours.
12. The method of treating bioprosthetic tissue of claim 1, wherein the capping agent is selected from at least one of amino acids, saturated alkyl amines, unsaturated alkyl amines, amino alcohols, polyether amines, amino alkyl acids, amino alkyl diacids, amino surfactants.
13. The method of treating bioprosthetic tissue of claim 1, wherein after the capping treatment, the method further comprises: and cleaning the bioprosthesis tissue for the second time by using a second cleaning solution, wherein the second cleaning solution is one selected from an ethanol solution, normal saline or a phosphate buffer solution, the number of times of cleaning for the second time is 3-5, and each time of cleaning is 3-5 min.
14. A bioprosthetic heart valve comprising bioprosthetic tissue treated by the method of treating a bioprosthetic component according to any one of claims 1 to 13.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011573018.2A CN114652893A (en) | 2020-12-23 | 2020-12-23 | Method for treating bioprosthetic tissue and bioprosthetic heart valve |
| AU2021407693A AU2021407693A1 (en) | 2020-12-23 | 2021-10-29 | Biological prosthetic tissue processing method, and biological prosthetic heart valve |
| PCT/CN2021/127694 WO2022134858A1 (en) | 2020-12-23 | 2021-10-29 | Biological prosthetic tissue processing method, and biological prosthetic heart valve |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011573018.2A CN114652893A (en) | 2020-12-23 | 2020-12-23 | Method for treating bioprosthetic tissue and bioprosthetic heart valve |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114652893A true CN114652893A (en) | 2022-06-24 |
Family
ID=82025314
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011573018.2A Pending CN114652893A (en) | 2020-12-23 | 2020-12-23 | Method for treating bioprosthetic tissue and bioprosthetic heart valve |
Country Status (3)
| Country | Link |
|---|---|
| CN (1) | CN114652893A (en) |
| AU (1) | AU2021407693A1 (en) |
| WO (1) | WO2022134858A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024001217A1 (en) * | 2022-06-28 | 2024-01-04 | 江苏臻亿医疗科技有限公司 | Treatment method for biological tissue and medical material |
| WO2024078253A1 (en) * | 2022-10-12 | 2024-04-18 | 上海微创心通医疗科技有限公司 | Anti-calcification method for bioprosthetic tissue, and bioprosthetic tissue |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115337456B (en) * | 2022-08-08 | 2023-09-15 | 科凯(南通)生命科学有限公司 | High permeability solution and application thereof |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5697972A (en) * | 1994-07-13 | 1997-12-16 | Korea Institute Of Science And Technology | Bioprosthetic heart valves having high calcification resistance |
| WO1999022779A1 (en) * | 1997-10-30 | 1999-05-14 | Mitroflow International Inc. | Borohydride reduction of biological tissues |
| US20060217804A1 (en) * | 2005-03-25 | 2006-09-28 | Dove Jeffrey S | Treatment of bioprosthetic tissues to mitigate post implantation calcification |
| CN101818140A (en) * | 2010-04-28 | 2010-09-01 | 湖南大学 | Immobilization pseudomonas aeruginosa as well as preparation method and application thereof |
| CN101965205A (en) * | 2007-12-21 | 2011-02-02 | 爱德华兹生命科学公司 | Capping Bioprosthetic Tissue to Reduce Calcification |
| CN102737786A (en) * | 2012-06-28 | 2012-10-17 | 北京理工大学 | Preparation method of cellulose nanofiber-based flexible transparent conductive membrane |
| CN102946916A (en) * | 2010-06-17 | 2013-02-27 | 爱德华兹生命科学公司 | Methods for stabilizing a bioprosthetics tissue by chemical modification of antigenic carbohydrates |
| CN103313735A (en) * | 2010-11-17 | 2013-09-18 | 爱德华兹生命科学公司 | Double cross-linkage process to enhance post-implantation bioprosthetic tissue durability |
| CN106190949A (en) * | 2015-05-08 | 2016-12-07 | 上海微创心通医疗科技有限公司 | A kind of dry state animal derived collagenous tissue material and preparation method thereof and bioprosthesis |
| US20170252280A1 (en) * | 2014-10-15 | 2017-09-07 | Henkel Ag & Co. Kgaa | Method for simultaneously and permanently reshaping and coloring keratin fibers |
| CN108410913A (en) * | 2018-01-31 | 2018-08-17 | 杨妮 | The method and purposes of carrot tissue or callus asymmetric reduction p- phenylpropyl alcohol ketone class compounds |
| CN110366398A (en) * | 2017-03-06 | 2019-10-22 | 爱德华兹生命科学公司 | Moisture management packaging system and method |
| CN111217708A (en) * | 2020-02-21 | 2020-06-02 | 中国林业科学研究院林产化学工业研究所 | Preparation method and weeding application of alkyl perillamine derivative |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107441556B (en) * | 2017-07-05 | 2020-07-17 | 北京大清生物技术股份有限公司 | Polyamino acid-terminated tissue repair material and preparation method thereof |
-
2020
- 2020-12-23 CN CN202011573018.2A patent/CN114652893A/en active Pending
-
2021
- 2021-10-29 WO PCT/CN2021/127694 patent/WO2022134858A1/en active Application Filing
- 2021-10-29 AU AU2021407693A patent/AU2021407693A1/en active Pending
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5697972A (en) * | 1994-07-13 | 1997-12-16 | Korea Institute Of Science And Technology | Bioprosthetic heart valves having high calcification resistance |
| WO1999022779A1 (en) * | 1997-10-30 | 1999-05-14 | Mitroflow International Inc. | Borohydride reduction of biological tissues |
| US20060217804A1 (en) * | 2005-03-25 | 2006-09-28 | Dove Jeffrey S | Treatment of bioprosthetic tissues to mitigate post implantation calcification |
| CN101965205A (en) * | 2007-12-21 | 2011-02-02 | 爱德华兹生命科学公司 | Capping Bioprosthetic Tissue to Reduce Calcification |
| CN103705980A (en) * | 2007-12-21 | 2014-04-09 | 爱德华兹生命科学公司 | Clutch suitable for vehicles' powered mirrors |
| CN101818140A (en) * | 2010-04-28 | 2010-09-01 | 湖南大学 | Immobilization pseudomonas aeruginosa as well as preparation method and application thereof |
| CN102946916A (en) * | 2010-06-17 | 2013-02-27 | 爱德华兹生命科学公司 | Methods for stabilizing a bioprosthetics tissue by chemical modification of antigenic carbohydrates |
| CN103313735A (en) * | 2010-11-17 | 2013-09-18 | 爱德华兹生命科学公司 | Double cross-linkage process to enhance post-implantation bioprosthetic tissue durability |
| CN102737786A (en) * | 2012-06-28 | 2012-10-17 | 北京理工大学 | Preparation method of cellulose nanofiber-based flexible transparent conductive membrane |
| US20170252280A1 (en) * | 2014-10-15 | 2017-09-07 | Henkel Ag & Co. Kgaa | Method for simultaneously and permanently reshaping and coloring keratin fibers |
| CN106190949A (en) * | 2015-05-08 | 2016-12-07 | 上海微创心通医疗科技有限公司 | A kind of dry state animal derived collagenous tissue material and preparation method thereof and bioprosthesis |
| CN110366398A (en) * | 2017-03-06 | 2019-10-22 | 爱德华兹生命科学公司 | Moisture management packaging system and method |
| CN108410913A (en) * | 2018-01-31 | 2018-08-17 | 杨妮 | The method and purposes of carrot tissue or callus asymmetric reduction p- phenylpropyl alcohol ketone class compounds |
| CN111217708A (en) * | 2020-02-21 | 2020-06-02 | 中国林业科学研究院林产化学工业研究所 | Preparation method and weeding application of alkyl perillamine derivative |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024001217A1 (en) * | 2022-06-28 | 2024-01-04 | 江苏臻亿医疗科技有限公司 | Treatment method for biological tissue and medical material |
| WO2024078253A1 (en) * | 2022-10-12 | 2024-04-18 | 上海微创心通医疗科技有限公司 | Anti-calcification method for bioprosthetic tissue, and bioprosthetic tissue |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2022134858A1 (en) | 2022-06-30 |
| AU2021407693A1 (en) | 2023-06-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114652893A (en) | Method for treating bioprosthetic tissue and bioprosthetic heart valve | |
| US7579381B2 (en) | Treatment of bioprosthetic tissues to mitigate post implantation calcification | |
| US20060217804A1 (en) | Treatment of bioprosthetic tissues to mitigate post implantation calcification | |
| US10383978B2 (en) | Dry animal-derived collagen fiber tissue material and preparation method and bioprothesis thereof | |
| EP3854425B1 (en) | Pre-loadable dried biological heart valve and preparation method thereof | |
| WO2018167536A1 (en) | Implantable material and method for preserving | |
| CN113499478B (en) | Method for processing biological tissue material | |
| EP3693030A1 (en) | Method for preparing biological tissue for surgical implantation | |
| CN107802889A (en) | A kind of animal derived collagenous tissue material of dry state and preparation method thereof | |
| US20240075187A1 (en) | Biological tissue material and preparation method therefor | |
| CN112755247B (en) | Acellular dermal matrix and preparation method thereof | |
| CN111701079A (en) | Biological valve and treatment method thereof | |
| WO2022143700A1 (en) | Fold resistant dehydrated cross-linked biological material, preparation method therefor, and application thereof | |
| US20220176018A1 (en) | A process for prevention of degradation and degeneration of tissue used in bioprosthesis | |
| US5120833A (en) | Method of producing grafts | |
| CN111110920B (en) | Biological patch and preparation method thereof | |
| CN105770991A (en) | Preparing method of biological source venous valve | |
| CN109820624B (en) | Method for treating biological valve by adopting photo-crosslinking | |
| CN110917398A (en) | Method for oxidation resistance of biomaterial and biomaterial | |
| US20190374680A1 (en) | Method for the preparation of biological tissue for dry use in an implant | |
| WO2020038293A1 (en) | Method for treating biological valve using combination of enzymatic cross-linking and tea polyphenol | |
| CN112236175B (en) | Method for preparing biological tissue for surgical implantation | |
| US20200171205A1 (en) | Collagen fibers and articles formed therefrom | |
| WO2024078253A1 (en) | Anti-calcification method for bioprosthetic tissue, and bioprosthetic tissue | |
| WO2024114050A1 (en) | Biological tissue material, preparation method therefor, and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |