WO2020057412A1 - 一种可预装干燥生物心脏瓣膜及其制备方法 - Google Patents

一种可预装干燥生物心脏瓣膜及其制备方法 Download PDF

Info

Publication number
WO2020057412A1
WO2020057412A1 PCT/CN2019/105345 CN2019105345W WO2020057412A1 WO 2020057412 A1 WO2020057412 A1 WO 2020057412A1 CN 2019105345 W CN2019105345 W CN 2019105345W WO 2020057412 A1 WO2020057412 A1 WO 2020057412A1
Authority
WO
WIPO (PCT)
Prior art keywords
heart valve
pericardium
biological heart
cross
preparing
Prior art date
Application number
PCT/CN2019/105345
Other languages
English (en)
French (fr)
Inventor
王云兵
雷洋
李高参
杨立
林浩昇
邝大军
余金城
Original Assignee
杭州启明医疗器械股份有限公司
四川大学
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 杭州启明医疗器械股份有限公司, 四川大学 filed Critical 杭州启明医疗器械股份有限公司
Priority to EP19862226.8A priority Critical patent/EP3854425B1/en
Publication of WO2020057412A1 publication Critical patent/WO2020057412A1/zh
Priority to US17/313,877 priority patent/US20210260247A1/en

Links

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
    • A61L27/3625—Vascular tissue, 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
    • 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
    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
    • 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

Definitions

  • the invention relates to the field of medical materials and medical devices, and in particular, to a pre-loaded dry biological heart valve and a preparation method thereof.
  • Heart valve disease is a common valvular disease. Anatomically manifested as narrowed blood passages or valvular insufficiency. Treatment of heart valve disease includes thoracotomy and percutaneous heart valve replacement. Thoracic surgery is very traumatic, high risk, slow recovery, and requires extracorporeal circulation support, which is unacceptable to many patients. Percutaneous heart valve replacement surgery has become the main trend of future valve surgery because it has less trauma to patients and low risk. Biological heart valves are a class of biomedical materials used to replace heart valves in humans. Biological heart valves are generally prepared by cross-linking glutaraldehyde from porcine pericardium and bovine pericardium.
  • the existing interventional biological valve is generally cross-linked by glutaraldehyde, and then the biological valve is sewn on a metal stent, and then stored in a glutaraldehyde solution.
  • glutaraldehyde solution Prior to interventional valve replacement surgery, multiple washing, crimping, and assembly steps are required. This preparation process is cumbersome and easily leads to an increased risk of surgery.
  • the biological valve stored in the glutaraldehyde solution for a long time can easily lead to the residual of glutaraldehyde and increase the calcification and toxicity of the biological valve.
  • the preservation of glutaraldehyde makes it necessary to wash it when it is used at the operation site. After washing, it is compressed and loaded into the delivery system at the operation site, which substantially increases the operation time.
  • the development of the biological valve into a dry film stored away from the glutaraldehyde solution, and pre-compressed and loaded in the valve delivery system can better solve the above problems.
  • the dry biological valve pre-loaded in the valve delivery system is subject to a longer period of mechanical folding and is prone to folding damage. Therefore, higher requirements are placed on the toughness of the biological valve under the dry loading state. Existing biological valve toughness is still insufficient.
  • an object of the present invention is to provide a prefilled dry biological heart valve and a preparation method thereof, which aims to solve the need for the existing interventional biological valve to be stored in a glutaraldehyde solution and cannot be pre-loaded in In the conveying system, the problems of temporary cleaning and clamping assembly and insufficient toughness are required.
  • the invention provides a method for preparing a dry biological heart valve, which includes:
  • Step A After immersing the fresh animal pericardium in an aqueous solution of soluble elastin or glycosaminoglycan, perform a cross-linking reaction in a carbodiimide / N-hydroxysuccinimide mixed solution to elastin or Glycosaminoglycans are bound to the pericardium via chemical bonds; and
  • Step B The pericardial membrane after the primary cross-linking is subjected to a secondary cross-linking reaction with an glutaraldehyde aqueous solution, and then the pericardial membrane after the secondary cross-linking is dehydrated and dried to obtain a pre-dried biological heart valve.
  • Elastin or glycosaminoglycan contains a carboxyl group that can react with amino groups in fresh pericardium, under the conditions of carbodiimide / N-hydroxysuccinimide mixed solution, chemical cross-linking reaction occurs and dehydration condensation forms amide bonds Therefore, the elastin or glycosaminoglycan is bound to the pericardium through chemical bonds, which improves the toughness of the biological valve, and avoids mechanical folding caused by compression and loading for a long time.
  • the pericardium is in an aqueous solution of soluble elastin or glycosaminoglycan with a mass concentration of 0.1 to 10% at 25 to 37 Soak for 2 to 24 hours under shaking at °C.
  • the method for preparing a preloaded dry biological heart valve wherein in the step A, the pericardium is in a carbodiimide / N-hydroxysuccinimide mixed solution with a molar concentration of 0.1 to 1M. After soaking in a shaking condition at 25 to 37 ° C for 2 to 24 hours, a crosslinking reaction was performed.
  • the pericardial membrane after one-time cross-linking is immersed in a glutaraldehyde aqueous solution with a volume concentration of 0.25 to 1% in 25 to 37
  • the cross-linking reaction was carried out for 1 to 7 days under shaking conditions at °C.
  • the drying is dehydration drying in an alcohol solvent.
  • the method for preparing a pre-loaded dry biological heart valve wherein in the step B, the pericardial membrane after secondary cross-linking is dehydrated and dried in a gradient concentration in an alcohol solvent.
  • the alcohol solvents include methanol, ethanol, isopropanol, glycerol, n-propanol, n-butanol, and n-pentyl.
  • the alcohol solvents include methanol, ethanol, isopropanol, glycerol, n-propanol, n-butanol, and n-pentyl.
  • the method for preparing a preloaded dry biological heart valve wherein in the step B, the pericardial membrane after the second cross-linking is sequentially immersed in 50% isopropanol / 50% propylene at 25-37 ° C. Triol is 20-30h, 90% isopropanol / 10% glycerol is dried for 20-30h.
  • the soluble elastin or glycosaminoglycan solution is soluble elastin, hyaluronic acid, chondroitin sulfate, dermatan sulfate, sulfuric acid
  • the soluble elastin or glycosaminoglycan solution is soluble elastin, hyaluronic acid, chondroitin sulfate, dermatan sulfate, sulfuric acid
  • the method for preparing a pre-loaded dry biological heart valve wherein in the step A, immersing the fresh animal pericardium in an aqueous solution of soluble elastin or glycosaminoglycan also includes immersing the fresh animal pericardium Cleaning steps.
  • the method for preparing a pre-loaded dry biological heart valve wherein the step of cleaning the fresh animal pericardium includes: oscillating the fresh animal pericardium at a rotation speed of 3 to 5 ° C and a rotation speed of 80 to 120 RPM. Wash with distilled water for 1.5 ⁇ 3h.
  • the method for preparing a pre-loaded dry biological heart valve, wherein the step of washing the pericardium of fresh animals clean includes a decellularization process.
  • the invention also provides a pre-loaded dry biological heart valve, which is prepared by using the preparation method described in any one of the above.
  • the present invention provides a pre-installed dry biological heart valve and a preparation method thereof.
  • the dry biological heart valve prepared by using the preparation method of the present invention has good toughness and can be quickly flattened under a simulated folding test. .
  • the present invention provides a pre-installed dry biological heart valve and a preparation method thereof.
  • the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.
  • a method for preparing a pre-loaded dry biological heart valve provided by a preferred embodiment of the present invention, that is, a method for toughening a pre-loaded dry biological heart valve, including:
  • Step S100 After immersing the fresh animal pericardium in an aqueous solution of soluble elastin or glycosaminoglycan, a cross-linking reaction is performed in a carbodiimide / N-hydroxysuccinimide mixed solution to elastin or Glycosaminoglycans are bound to the pericardium via chemical bonds; and
  • step S200 the pericardial membrane after the primary cross-linking is subjected to a secondary cross-linking reaction with an glutaraldehyde aqueous solution, and then the pericardial membrane after the secondary cross-linking is dried to obtain a preloaded dry biological heart valve.
  • the invention creatively permeates the pericardium of fresh animals by immersing them in an aqueous solution of water-soluble elastin or glycosaminoglycan, and the carboxyl group contained in elastin or glycosaminoglycan can be combined with the amino group in the fresh pericardium.
  • a mixed solution of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) / N-hydroxysuccinimide (NHS) a chemical crosslinking reaction occurs and dehydration condensation forms an amide bond. Therefore, the elastin or glycosaminoglycan is bound to the pericardium through chemical bonds, thereby improving the toughness of the biological valve.
  • the drying is preferably dehydrated and dried in an alcohol solvent, and more preferably, the pericardial membrane after the secondary crosslinking is dehydrated and dried in a gradient concentration in the alcohol solvent.
  • the preparation method of the present invention includes: 1. toughening chemical treatment of the biological valve; 2. cross-linking of the biological valve; 3. drying, and the drying is preferably dehydration drying in an alcohol solvent.
  • the toughening chemical treatment of biological valves and the dehydration and drying of gradient concentration alcohol solvents are the key steps of the present invention.
  • the animal pericardium according to the present invention includes, but is not limited to, a pig pericardial valve, a bovine pericardial valve, and the like.
  • the method before immersing the fresh animal pericardium in an aqueous solution of soluble elastin or glycosaminoglycan, the method further includes a step of cleaning the fresh animal pericardium, and the fresh animal pericardium is cleaned.
  • the steps include a decellularization process. During the specific implementation of the cleaning process, fresh pericardial tissues of pigs or cows were collected and stored under low temperature and humid conditions at 4 ° C.
  • the pericardial tissues were washed with distilled water under gentle friction and fluid pressure under oscillating conditions to remove adhered non-pericardial and Non-collagenous tissue.
  • the washing according to the present invention achieves effective decellularization of the pericardial tissue through osmotic shock, and preferably the washing is continued until there is no visible adhesion of non-pericardial or non-collagenous tissue.
  • the step of cleaning the fresh animal pericardium includes: washing the fresh animal pericardium with distilled water at a temperature of 3 to 5 ° C. and a rotation speed of 80 to 120 RPM for 1.5 to 3 hours.
  • the pericardium of fresh animal pig or bovine
  • the step S100 is a toughening chemical treatment process of the biological valve, and the soluble elastin or glycosaminoglycan solution is soluble elastin, hyaluronic acid, chondroitin sulfate, dermatan sulfate, keratan sulfate, acetyl sulfate One or more mixed solutions of heparin and heparin.
  • the pericardium is immersed in an aqueous solution of soluble elastin or glycosaminoglycan having a mass concentration of 0.1 to 10% under shaking conditions at 25 to 37 ° C. for 2 to 24 hours. This step needs to ensure that the water-soluble elastin or glycosaminoglycan reaches a physical saturation close to saturation, so that as much water-soluble elastin or glycosaminoglycan is introduced as possible.
  • the pericardium is immersed in a carbodiimide / N-hydroxysuccinimide mixed solution having a molar concentration of 0.1 to 1M under a shaking condition of 25 to 37 ° C for 2 to A crosslinking reaction was performed at 24 h.
  • This step will achieve stable chemical cross-linking between the water-soluble elastin or glycosaminoglycan that penetrated in the previous step and the pericardial tissue, thereby improving the toughness of the biological valve.
  • the pericardial membrane after being cross-linked once is immersed in a glutaraldehyde aqueous solution having a volume concentration of 0.25 to 1% and subjected to a cross-linking reaction under shaking conditions at 25 to 37 ° C for 1 to 7 days.
  • This step will achieve stable cross-linking of most collagen tissues, improve the structural stability of the entire pericardial tissue, and reduce or even eliminate immunogenicity.
  • the alcohol solvents include methanol, ethanol, isopropanol, glycerol, n-propanol, n-butanol, n-pentanol, n-undecanol, and n-dodecanol.
  • the alcohol solvent of the present invention may be a mixed solvent, and may be, for example, 50% isopropanol / 50% glycerol, 90% isopropanol / 10% glycerol, or the like.
  • the pericardial membrane after the second cross-linking is sequentially immersed in 50% isopropanol / 50% glycerol at 25 to 37 ° C for 20 to 30 hours (for example, 24 hours), 90% isopropanol / 10% propane.
  • Triol 20-30h (for example, 24h), realizes the gradient dehydration of the biological valve, thereby completing the drying of the biological heart valve.
  • the embodiment of the present invention also provides a pre-loaded dry biological heart valve, which is prepared by using the preparation method described above.
  • the dried biological heart valve prepared by using the preparation method of the present invention has good toughness and can be quickly flattened under a simulated folding test.
  • the prepared dry biological heart valve has good toughness and can be quickly flattened under a simulated bending test.
  • Pre-packed dry biovalves stored without glutaraldehyde solution. It can reduce the residue of glutaraldehyde, reduce the calcification problem and toxicity caused by glutaraldehyde, simplify the preoperative installation of the valve system, and reduce the additional risk of surgery.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Water-soluble elastin was from Sigma-Aldrich.
  • the cleaned pig pericardium was immersed in a 5% water-soluble elastin solution for 24 hours, removed and rinsed, and then 0.2M carbodiimide (EDC) /0.1M N-hydroxysuccinimide (NHS ) Cross-linking treatment of the mixed aqueous solution for 2 h and washing with distilled water.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Hyaluronic acid comes from Shanghai Aladdin Biochemical Technology Co., Ltd. Soak the cleaned pig pericardium in a 5% aqueous solution of hyaluronic acid for 24 hours, remove and rinse, then use 0.2M carbodiimide (EDC) /0.1M N-hydroxysuccinimide (NHS) The mixed aqueous solution was cross-linked for 2 h and washed with distilled water.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Water-soluble elastin was from Sigma-Aldrich.
  • Hyaluronic acid comes from Shanghai Aladdin Biochemical Technology Co., Ltd.
  • the cleaned pig pericardium was immersed in a mixed aqueous solution of 2.5% water-soluble elastin and 2.5% hyaluronic acid for 24 hours, removed and rinsed, and then 0.2M carbodiimide (EDC) /0.1M N-
  • EDC carbodiimide
  • NHS hydroxysuccinimide
  • the hydroxysuccinimide (NHS) mixed aqueous solution was cross-linked for 2 h and washed with distilled water. It was then cross-linked with a 0.5% aqueous solution of glutaraldehyde (pH 7.4, 25 ° C) for 72 h. Wash with distilled water.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Water-soluble elastin was from Sigma-Aldrich.
  • Hyaluronic acid comes from Shanghai Aladdin Biochemical Technology Co., Ltd.
  • the cleaned pig pericardium was immersed in a 0.1% water-soluble elastin solution for 24 hours, removed and rinsed, and then 0.2M carbodiimide (EDC) /0.1M N-hydroxysuccinimide (NHS ) Cross-linking treatment of the mixed aqueous solution for 2 h and washing with distilled water.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Water-soluble elastin was from Sigma-Aldrich.
  • Hyaluronic acid comes from Shanghai Aladdin Biochemical Technology Co., Ltd. Soak the cleaned pig pericardium in a 10% water-soluble elastin solution for 24 hours, remove and rinse, then use 0.2M carbodiimide (EDC) /0.1M N-hydroxysuccinimide (NHS ) Cross-linking treatment of the mixed aqueous solution for 2 h and washing with distilled water.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • Fresh pig pericardium comes from the local slaughterhouse.
  • Glutaraldehyde, carbodiimide (EDC), N-hydroxysuccinimide (NHS) were from Chengdu Best Reagent Company.
  • Water-soluble elastin was from Sigma-Aldrich.
  • Hyaluronic acid comes from Shanghai Aladdin Biochemical Technology Co., Ltd.
  • the dried biological heart valve prepared in this implementation can be quickly flattened within five minutes under the condition of simulated folding test (10kg weight folding after two folds), with good toughness and no obvious folding damage. . Under the uniaxial tensile test (1 * 5cm sample size), the tensile breaking stress is greater than 15N.
  • the present invention provides a pre-installed dry biological heart valve and a preparation method thereof.
  • the dried biological heart valve prepared by using the processing method of the present invention has good toughness and can quickly develop under simulated bending test. level.
  • the pre-installed dry biological valve stored without glutaraldehyde solution can reduce the residual glutaraldehyde, reduce the calcification problem and toxicity caused by glutaraldehyde, simplify the preoperative installation of the valve system, and reduce the additional risk of surgery.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Veterinary Medicine (AREA)
  • Botany (AREA)
  • Public Health (AREA)
  • Dermatology (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Materials For Medical Uses (AREA)
  • Prostheses (AREA)

Abstract

一种可预装干燥生物心脏瓣膜及其制备方法，制备方法包括：步骤A、将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡后，在碳二亚胺/N-羟基丁二酰亚胺混合溶液中进行一次交联反应，将弹性蛋白或糖胺多糖通过化学键结合在心包膜上；以及步骤B、将经过一次交联后的心包膜采用戊二醛水溶液进行二次交联反应，之后将二次交联后的心包膜干燥，得到可预装干燥生物心脏瓣膜。采用以上制备方法制得的干燥生物心脏瓣膜具有良好的韧性，在模拟折压测试下可以快速展平。

Description

一种可预装干燥生物心脏瓣膜及其制备方法 技术领域
本发明涉及医学材料及医疗器械领域，尤其涉及一种可预装干燥生物心脏瓣膜及其制备方法。
背景技术
心脏瓣膜疾病是一种常见的瓣膜衰退疾病。在解剖学上表现为血液通路变窄或瓣膜关闭不全。心脏瓣膜疾病的治疗包括开胸瓣膜置换手术以及经皮心脏瓣膜置换手术。开胸手术对病人创伤大、风险高、恢复慢、需体外循环支持，很多患者无法接受。经皮心脏瓣膜置换手术因为对病人创伤小、风险低，成为未来瓣膜手术的主要趋势。生物心脏瓣膜是指一类用于替换人体病变心脏瓣膜的生物医学材料。生物心脏瓣膜一般由猪心包膜、牛心包膜等通过戊二醛交联制备而成。
现有的介入生物瓣膜一般采用戊二醛交联，然后将生物瓣膜缝制于一种金属支架上，随后保存于戊二醛溶液当中。在进行介入瓣膜置换手术之前，需要经过多次清洗、压握和组装。这个准备过程繁琐，容易导致手术附加风险增大。长期存放于戊二醛溶液当中的生物瓣膜，容易导致戊二醛的残留，增加生物瓣膜的钙化和毒性。而且戊二醛的保存，使得在手术现场使用时，需要经过清洗，清洗后，在手术现场进行压缩装载到输送系统中，这无形中增加了手术的时间。因此，将生物瓣膜开发成为脱离戊二醛溶液保存的干膜，并预先压缩装载于瓣膜输送系统当中，可以较好解决上述问题。但是，预先装载于瓣膜输送系统当中的干燥生物瓣膜，由于要经受较长时间的机械压折，容易存在压折损伤，所以对干燥装载状态之下的生物瓣膜的韧性提出了更高的要求，现有的生物瓣膜韧性仍然不足。
因此，现有技术还有待于改进和发展。
发明内容
鉴于上述现有技术的不足，本发明的目的在于提供一种可预装干燥生物心脏瓣膜及其制备方法，旨在解决现有的介入生物瓣膜需要保存在戊二醛溶液当中、无法预先装载于输送系统当中、需要临时清洗压握组装以及韧性不足的问题。
本发明的技术方案如下：
本发明提供了一种可预装干燥生物心脏瓣膜的制备方法，其中，包括：
步骤A、将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡后，在碳二亚胺/N-羟基丁二酰亚胺混合溶液中进行一次交联反应，将弹性蛋白或糖胺多糖通过化学键结合在心包膜上；以及
步骤B、将经过一次交联后的心包膜采用戊二醛水溶液进行二次交联反应，之后将二次交联后的心包膜脱水干燥，得到可预装干燥生物心脏瓣膜。
弹性蛋白或糖胺多糖所含羧基可以和新鲜心包膜中的氨基，在碳二亚胺/N-羟基丁二酰亚胺混合溶液作用条件下，发生化学交联反应而脱水缩合形成酰胺键，从而将弹性蛋白或糖胺多糖通过化学键结合在心包膜上，提高了生物瓣膜的韧性，避免了压握装载后，长时间处于压缩状态而导致的机械压折。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤A中，将心包膜在质量浓度为0.1～10％的可溶性弹性蛋白或糖胺多糖的水溶液中，于25～37℃的振荡条件下浸泡2～24h。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤A中，将心包膜在摩尔浓度为0.1～1M的碳二亚胺/N-羟基丁二酰亚胺混合溶液中，于25～37℃的振荡条件下浸泡2～24h进行一次交联反应。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤B中，将经过一次交联后的心包膜浸泡于体积浓度为0.25～1％的戊二醛水溶液于 25～37℃的振荡条件下交联反应1～7天。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤B中，所述干燥是在醇类溶剂中脱水干燥。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤B中，二次交联后的心包膜在醇类溶剂中完成梯度浓度脱水干燥。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤B中，所述醇类溶剂包括甲醇、乙醇、异丙醇、丙三醇、正丙醇、正丁醇、正戊醇、正十一烷醇、正十二烷醇、2-丙醇、2-丁醇、2-己醇、环己醇、叔丁醇的一种或多种。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤B中，将二次交联后的心包膜在25～37℃下依次浸泡于50％异丙醇/50％丙三醇20～30h，90％异丙醇/10％丙三醇20～30h完成脱水干燥。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤A中，所述可溶性弹性蛋白或糖胺多糖溶液为可溶性弹性蛋白、透明质酸、硫酸软骨素、硫酸皮肤素、硫酸角质素、硫酸乙酰肝素和肝素中的一种或多种混合溶液。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述步骤A中，将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡前，还包括将新鲜动物心包膜清洗干净的步骤。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述将新鲜动物心包膜清洗干净的步骤包括：将新鲜动物心包膜在3～5℃、80～120RPM的转速振荡条件之下以蒸馏水清洗1.5～3h。
所述的可预装干燥生物心脏瓣膜的制备方法，其中，所述将新鲜动物心包膜清洗干净的步骤包括脱细胞处理工序。
本发明还提供了一种可预装干燥生物心脏瓣膜，其中，采用如以上任一项所述的制备方法制得。
有益效果：本发明提供了一种可预装干燥生物心脏瓣膜及其制备方法，采用本发明所述制备方法制得的干燥生物心脏瓣膜具有良好的韧性，在模拟折压测试下可以快速展平。
具体实施方式
本发明提供一种可预装干燥生物心脏瓣膜及其制备方法，为使本发明的目的、技术方案及效果更加清楚、明确，以下对本发明进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
本发明较佳实施例提供的一种可预装干燥生物心脏瓣膜的制备方法，也即是一种可预装干燥生物心脏瓣膜的增韧处理方法，其包括：
步骤S100、将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡后，在碳二亚胺/N-羟基丁二酰亚胺混合溶液中进行一次交联反应，将弹性蛋白或糖胺多糖通过化学键结合在心包膜上；以及
步骤S200、将经过一次交联后的心包膜采用戊二醛水溶液进行二次交联反应，之后将二次交联后的心包膜干燥，得到可预装干燥生物心脏瓣膜。
本发明创造性的将新鲜动物心包膜，通过浸泡于水溶性弹性蛋白或糖胺多糖的水溶液中后，弹性蛋白或糖胺多糖所含羧基可以和新鲜心包膜中的氨基，在碳二亚胺(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide，EDC)/N-羟基丁二酰亚胺(N-hydroxysuccinimide，NHS)混合溶液作用条件下，发生化学交联反应而脱水缩合形成酰胺键，从而将弹性蛋白或糖胺多糖通过化学键结合在心包膜上，提高了生物瓣膜的韧性。
所述步骤S200中，所述干燥优选是在醇类溶剂中脱水干燥，更优选的，二次交联后的心包膜在醇类溶剂中完成梯度浓度脱水干燥。
整体来说，本发明所述制备方法包括：1.生物瓣膜的增韧化学处理；2.生物瓣膜的交联；3.干燥，所述干燥优选为在醇类溶剂中脱水干燥。其中，生物瓣膜的增韧化学处理和梯度浓度醇类溶剂的脱水干燥是本发明的 关键步骤。
进一步的，本发明所述动物心包膜包括但不限于猪心包瓣膜、牛心包瓣膜等。所述步骤S100中，将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡前，还包括将新鲜动物心包膜清洗干净的步骤，所述将新鲜动物心包膜清洗干净的步骤包括脱细胞处理工序。清洗过程具体实施时，采集新鲜的猪或牛的心包组织，并于4℃低温湿润状态下保存，采用柔和摩擦和流体压力在振荡条件之下用蒸馏水清洗心包组织，去除粘附的非心包和非胶原组织。本发明所述清洗通过渗压休克实现对心包组织有效脱细胞，优选清洗持续到没有可见的粘附的非心包或非胶原组织。具体实施时，优选的，所述将新鲜动物心包膜清洗干净的步骤包括：将新鲜动物心包膜在3～5℃、80～120RPM的转速振荡条件之下以蒸馏水清洗1.5～3h。例如，可以在4℃100RPM转速振荡条件之下蒸馏水清洗2h，然后将清洗干净的新鲜动物(猪或牛)心包膜浸入可溶性弹性蛋白或是糖胺多糖的水溶液中。
进一步的，所述步骤S100为生物瓣膜的增韧化学处理过程，所述可溶性弹性蛋白或糖胺多糖溶液为可溶性弹性蛋白、透明质酸、硫酸软骨素、硫酸皮肤素、硫酸角质素、硫酸乙酰肝素和肝素中的一种或多种混合溶液。优选的，将心包膜在质量浓度为0.1～10％的可溶性弹性蛋白或糖胺多糖的水溶液中，于25～37℃的振荡条件下浸泡2～24h。此步骤需要确保水溶性弹性蛋白或糖胺多糖达到接近饱和的物理渗透，从而尽可能多地引入水溶性弹性蛋白或糖胺多糖。
进一步的，所述步骤S100中，将心包膜在摩尔浓度为0.1～1M的碳二亚胺/N-羟基丁二酰亚胺混合溶液中，于25～37℃的振荡条件下浸泡2～24h进行一次交联反应。此步骤将实现前一步骤渗透的水溶性弹性蛋白或糖胺多糖与心包组织发生稳定的化学交联，从而提高生物瓣膜的韧性。
进一步的，所述步骤S200中，将经过一次交联后的心包膜浸泡于体积浓度为0.25～1％的戊二醛水溶液于25～37℃的振荡条件下交联反应1～7天。 此步骤将实现大部分胶原组织的稳定交联，提高整个心包组织的结构稳定性，降低以至于消除免疫原性。
进一步的，所述步骤S200中，所述醇类溶剂包括甲醇、乙醇、异丙醇、丙三醇、正丙醇、正丁醇、正戊醇、正十一烷醇、正十二烷醇、2-丙醇、2-丁醇、2-己醇、环己醇、叔丁醇的一种或多种。本发明醇类溶剂可以为混合溶剂，例如可以为50％异丙醇/50％丙三醇、90％异丙醇/10％丙三醇等。优选的，将二次交联后的心包膜在25～37℃下依次浸泡于50％异丙醇/50％丙三醇20～30h(例如24h)，90％异丙醇/10％丙三醇20～30h(例如24h)，实现生物瓣膜的梯度脱水，从而完成生物心脏瓣膜的干燥。
本发明实施例还提供了一种可预装干燥生物心脏瓣膜，其采用以上所述的制备方法制得。采用本发明所述制备方法制得的干燥生物心脏瓣膜具有良好的韧性，在模拟折压测试下可以快速展平。
本发明通过独特的干膜增韧处理技术，使得制备的干燥生物心脏瓣膜具有良好的韧性，在模拟折压测试下可以快速展平。脱离戊二醛溶液保存的可预装干燥生物瓣膜。可以减少戊二醛的残留，减少戊二醛带来的钙化问题以及毒性，简化瓣膜系统的术前安装，降低手术的附加风险。
下面以具体实施例对本发明做详细说明：
实施例1
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)来自于成都贝斯特试剂公司。水溶性弹性蛋白来自于Sigma-Aldrich公司。将清洗干净的猪心包膜在浓度为5％的水溶性弹性蛋白水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC)/0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇 24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
实施例2
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)自于成都贝斯特试剂公司。透明质酸来自于上海阿拉丁生化科技股份有限公司。将清洗干净的猪心包膜在浓度为5％的透明质酸水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC)/0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
实施例3
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)自于成都贝斯特试剂公司。水溶性弹性蛋白来自于Sigma-Aldrich公司。透明质酸来自于上海阿拉丁生化科技股份有限公司。将清洗干净的猪心包膜在浓度为2.5％的水溶性弹性蛋白以及2.5％的透明质酸混合水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC) /0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
实施例4
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)自于成都贝斯特试剂公司。水溶性弹性蛋白来自于Sigma-Aldrich公司。透明质酸来自于上海阿拉丁生化科技股份有限公司。将清洗干净的猪心包膜在浓度为0.1％的水溶性弹性蛋白水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC)/0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
实施例5
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)自于成都贝斯特试剂公司。水溶性弹性蛋白来自于 Sigma-Aldrich公司。透明质酸来自于上海阿拉丁生化科技股份有限公司。将清洗干净的猪心包膜在浓度为10％的水溶性弹性蛋白水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC)/0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
实施例6
新鲜猪心包膜来自当地屠宰场。戊二醛、碳二亚胺(EDC)、N-羟基丁二酰亚胺(NHS)自于成都贝斯特试剂公司。水溶性弹性蛋白来自于Sigma-Aldrich公司。透明质酸来自于上海阿拉丁生化科技股份有限公司。将清洗干净的猪心包膜在浓度为2.5％的水溶性弹性蛋白、透明质酸和硫酸软骨素水溶液中浸泡24h，取出冲洗，然后采用0.2M碳二亚胺(EDC)/0.1M N-羟基丁二酰亚胺(NHS)混合水溶液交联处理2h，蒸馏水清洗。然后采用0.5％戊二醛水溶液(pH 7.4，25℃)交联72h。蒸馏水清洗。然后在25℃条件之下依次浸泡于50％异丙醇/50％丙三醇24h，90％异丙醇/10％丙三醇24h，实现生物瓣膜的梯度脱水。制备得到具有良好韧性的可预装干燥生物心脏瓣膜。
经检测发现，本实施制备的干燥生物心脏瓣膜在模拟折压测试条件之下(对折两次之后10kg重物折压30天)能够在五分钟之内快速展平，韧性良好，无明显折叠损伤。单轴拉伸测试之下(1*5cm样品尺寸)拉伸断裂应力大于15N。
综上所述，本发明提供了一种可预装干燥生物心脏瓣膜及其制备方法，采用本发明所述处理方法制备的干燥生物心脏瓣膜具有良好的韧性，在模拟折压测试下可以快速展平。脱离戊二醛溶液保存的可预装干燥生物瓣膜，可以减少戊二醛的残留，减少戊二醛带来的钙化问题以及毒性，简化瓣膜系统的术前安装，降低手术的附加风险。
应当理解的是，本发明的应用不限于上述的举例，对本领域普通技术人员来说，可以根据上述说明加以改进或变换，所有这些改进和变换都应属于本发明所附权利要求的保护范围。

Claims (13)

  1. 一种可预装干燥生物心脏瓣膜的制备方法，其特征在于，包括：
    步骤A、将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡后，在碳二亚胺/N-羟基丁二酰亚胺混合溶液中进行一次交联反应，将弹性蛋白或糖胺多糖通过化学键结合在心包膜上；以及
    步骤B、将经过一次交联后的心包膜采用戊二醛水溶液进行二次交联反应，之后将二次交联后的心包膜干燥，得到可预装干燥生物心脏瓣膜。
  2. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤A中，将心包膜在质量浓度为0.1～10％的可溶性弹性蛋白或糖胺多糖的水溶液中，于25～37℃的振荡条件下浸泡2～24h。
  3. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤A中，将心包膜在摩尔浓度为0.1～1M的碳二亚胺/N-羟基丁二酰亚胺混合溶液中，于25～37℃的振荡条件下浸泡2～24h进行一次交联反应。
  4. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤B中，将经过一次交联后的心包膜浸泡于体积浓度为0.25～1％的戊二醛水溶液于25～37℃的振荡条件下交联反应1～7天。
  5. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤B中，所述干燥是在醇类溶剂中脱水干燥。
  6. 根据权利要求5所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤B中，二次交联后的心包膜在醇类溶剂中完成梯度浓度脱水干燥。
  7. 根据权利要求5所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤B中，所述醇类溶剂包括甲醇、乙醇、异丙醇、丙三醇、正丙醇、正丁醇、正戊醇、正十一烷醇、正十二烷醇、2-丙醇、2-丁醇、2-己醇、环己醇、叔丁醇的一种或多种。
  8. 根据权利要求6或7所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤B中，将二次交联后的心包膜在25～37℃下依次浸泡于50％异丙醇/50％丙三醇20～30h，90％异丙醇/10％丙三醇20～30h完成脱水干燥。
  9. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤A中，所述可溶性弹性蛋白或糖胺多糖溶液为可溶性弹性蛋白、透明质酸、硫酸软骨素、硫酸皮肤素、硫酸角质素、硫酸乙酰肝素和肝素中的一种或多种混合溶液。
  10. 根据权利要求1所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述步骤A中，将新鲜动物心包膜在可溶性弹性蛋白或糖胺多糖的水溶液中浸泡前，还包括将新鲜动物心包膜清洗干净的步骤。
  11. 根据权利要求10所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述将新鲜动物心包膜清洗干净的步骤包括：将新鲜动物心包膜在3～5℃、80～120RPM的转速振荡条件之下以蒸馏水清洗1.5～3h。
  12. 根据权利要求10所述的可预装干燥生物心脏瓣膜的制备方法，其特征在于，所述将新鲜动物心包膜清洗干净的步骤包括脱细胞处理工序。
  13. 一种可预装干燥生物心脏瓣膜，其特征在于，采用如权利要求1～12任一项所述的制备方法制得。
PCT/CN2019/105345 2018-09-19 2019-09-11 一种可预装干燥生物心脏瓣膜及其制备方法 WO2020057412A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP19862226.8A EP3854425B1 (en) 2018-09-19 2019-09-11 Pre-loadable dried biological heart valve and preparation method thereof
US17/313,877 US20210260247A1 (en) 2018-09-19 2021-05-06 Pre-Loadable Dried Biological Heart Valve and Preparation Method Thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201811096562.5 2018-09-19
CN201811096562.5A CN109260517B (zh) 2018-09-19 2018-09-19 一种可预装干燥生物心脏瓣膜及其制备方法

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US17/313,877 Continuation US20210260247A1 (en) 2018-09-19 2021-05-06 Pre-Loadable Dried Biological Heart Valve and Preparation Method Thereof

Publications (1)

Publication Number Publication Date
WO2020057412A1 true WO2020057412A1 (zh) 2020-03-26

Family

ID=65198247

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/105345 WO2020057412A1 (zh) 2018-09-19 2019-09-11 一种可预装干燥生物心脏瓣膜及其制备方法

Country Status (4)

Country Link
US (1) US20210260247A1 (zh)
EP (1) EP3854425B1 (zh)
CN (1) CN109260517B (zh)
WO (1) WO2020057412A1 (zh)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109260517B (zh) * 2018-09-19 2020-10-30 杭州启明医疗器械股份有限公司 一种可预装干燥生物心脏瓣膜及其制备方法
CN109821070A (zh) * 2019-02-22 2019-05-31 四川大学 一种可浸水快速展平的干燥生物心脏瓣膜及其制备方法
CN110101912B (zh) * 2019-05-16 2020-03-10 四川大学 一种含促进内皮化多肽的干燥人工生物心脏瓣膜及其制备方法
CN110101911B (zh) * 2019-05-16 2020-03-10 四川大学 一种含促进内皮化生长因子的干燥人工生物心脏瓣膜及其制备方法
CN110339400A (zh) * 2019-08-07 2019-10-18 太仓新亚逊生物科技有限公司 生物材料的抗钙化处理方法
CN110507856A (zh) * 2019-09-29 2019-11-29 四川大学 一种全预装干燥介入主动脉瓣膜材料及其制备方法
CN111567514A (zh) * 2020-04-20 2020-08-25 上海以心医疗器械有限公司 一种保存新鲜生物组织的方法
CN111658824B (zh) * 2020-06-15 2021-03-30 四川大学 一种具有协同抗凝及抗钙化功能的瓣膜材料及其制备方法
CN114681673B (zh) * 2020-12-31 2023-05-23 杭州启明医疗器械股份有限公司 抗折痕的脱水交联生物材料及其制备方法和应用
CN112957530B (zh) * 2021-02-05 2022-09-20 上海翰凌医疗器械有限公司 一种干燥生物组织材料及其制备方法

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5782931A (en) * 1996-07-30 1998-07-21 Baxter International Inc. Methods for mitigating calcification and improving durability in glutaraldehyde-fixed bioprostheses and articles manufactured by such methods
CN104334121A (zh) * 2012-06-04 2015-02-04 爱德华兹生命科学公司 预组装的生物假体瓣膜和密封导管
CN105194733A (zh) * 2014-06-30 2015-12-30 王静 一种化学处理人工瓣膜的方法
CN105326581A (zh) * 2015-09-29 2016-02-17 中国科学院金属研究所 一种制备聚乙二醇-蛋白质纤维复合人工心脏瓣膜的方法
CN105963783A (zh) * 2016-04-29 2016-09-28 陕西瑞盛生物科技有限公司 一种生物补片的交联方法以及交联生物补片
CN106913909A (zh) * 2017-02-27 2017-07-04 杭州启明医疗器械有限公司 一种干燥人工生物瓣膜及其制备方法
CN108785747A (zh) * 2018-07-07 2018-11-13 四川大学 一种提高生物材料抗钙化性能的处理方法及其生物材料
CN109260517A (zh) * 2018-09-19 2019-01-25 杭州启明医疗器械有限公司 一种可预装干燥生物心脏瓣膜及其制备方法

Family Cites Families (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4755593A (en) * 1985-07-24 1988-07-05 Lauren Mark D Novel biomaterial of cross-linked peritoneal tissue
US8067149B2 (en) * 1990-09-12 2011-11-29 Lifecell Corporation Acellular dermal matrix and method of use thereof for grafting
US6290991B1 (en) * 1994-12-02 2001-09-18 Quandrant Holdings Cambridge Limited Solid dose delivery vehicle and methods of making same
US6210440B1 (en) * 1995-09-15 2001-04-03 Kevin R. Stone Anterior cruciate ligament xenografts
US6193749B1 (en) * 1996-02-05 2001-02-27 St. Jude Medical, Inc. Calcification-resistant biomaterials
US20050163818A1 (en) * 1996-11-05 2005-07-28 Hsing-Wen Sung Drug-eluting device chemically treated with genipin
US6506339B1 (en) * 1997-02-10 2003-01-14 Biomedical Design, Inc. Method of sterilization
AU746318B2 (en) * 1997-04-11 2002-04-18 Cryolife, Inc. Tissue decellularization
US6254635B1 (en) * 1998-02-02 2001-07-03 St. Jude Medical, Inc. Calcification-resistant medical articles
US6383732B1 (en) * 1999-02-11 2002-05-07 Crosscart, Inc. Method of preparing xenograft heart valves
US6258320B1 (en) * 1998-04-09 2001-07-10 David H. Persing Method for preservation of nucleic acids
CA2335152C (en) * 1998-05-14 2008-11-18 The Cleveland Clinic Foundation Processing of implantable animal tissues for dry storage
US6933326B1 (en) * 1998-06-19 2005-08-23 Lifecell Coporation Particulate acellular tissue matrix
US6156531A (en) * 1998-07-20 2000-12-05 Sulzer Carbomedics Inc. Cross-linking tissue with a compound having a C8 to C40 aliphatic chain
US6177514B1 (en) * 1999-04-09 2001-01-23 Sulzer Carbomedics Inc. Blocked functional reagants for cross-linking biological tissues
US6132986A (en) * 1999-04-23 2000-10-17 Sulzer Carbomedics Inc. Tissue crosslinking for bioprostheses using activated difunctional or polyfunctional acids
US6312474B1 (en) * 1999-09-15 2001-11-06 Bio-Vascular, Inc. Resorbable implant materials
KR20010038098A (ko) * 1999-10-22 2001-05-15 박호군 헤파린 처리된 항석회화성 생체조직 이식물 및 이의 제조 방법
US6322994B1 (en) * 1999-11-04 2001-11-27 Genetix Limited Method of freeze-drying organisms
US6479079B1 (en) * 1999-12-13 2002-11-12 Sulzer Carbomedics Inc. Anticalcification treatments for fixed biomaterials
US6653062B1 (en) * 2000-07-26 2003-11-25 Wisconsin Alumni Research Foundation Preservation and storage medium for biological materials
US6696074B2 (en) * 2000-12-04 2004-02-24 Tei Biosciences, Inc. Processing fetal or neo-natal tissue to produce a scaffold for tissue engineering
GB2375771A (en) * 2001-05-24 2002-11-27 Univ Leeds Decellularisation of tissue implant material
US6878168B2 (en) * 2002-01-03 2005-04-12 Edwards Lifesciences Corporation Treatment of bioprosthetic tissues to mitigate post implantation calcification
US7189259B2 (en) * 2002-11-26 2007-03-13 Clemson University Tissue material and process for bioprosthesis
US20060177426A1 (en) * 2005-02-09 2006-08-10 Cecil Gibson Method of preserving lyophilized microorganisms for transport, storage and recovery of viable microorganisms
US20060204445A1 (en) * 2005-03-11 2006-09-14 Anthony Atala Cell scaffold matrices with image contrast agents
US7579381B2 (en) * 2005-03-25 2009-08-25 Edwards Lifesciences Corporation Treatment of bioprosthetic tissues to mitigate post implantation calcification
KR100739422B1 (ko) * 2005-04-26 2007-07-13 박기동 헤파린이 결합된 항석회화성 무세포성 생체조직 이식물 및그 제조방법
US20070292459A1 (en) * 2005-07-18 2007-12-20 Cooper Sarah M Halloysite microtubule processes, structures, and compositions
CN101626682B (zh) * 2006-10-27 2014-04-16 爱德华兹生命科学公司 用于外科植入的生物组织
US20080195230A1 (en) * 2007-02-09 2008-08-14 Quijano Rodolfo C Pericardial tissue sheet
US9101691B2 (en) * 2007-06-11 2015-08-11 Edwards Lifesciences Corporation Methods for pre-stressing and capping bioprosthetic tissue
US8357387B2 (en) * 2007-12-21 2013-01-22 Edwards Lifesciences Corporation Capping bioprosthetic tissue to reduce calcification
CN101721745B (zh) * 2009-11-09 2015-04-01 山东省千佛山医院 用于人工心脏瓣膜及其他生物修补材料的驴心包加工方法
CN101947147A (zh) * 2010-06-23 2011-01-19 乐普（北京）医疗器械股份有限公司 一种生物组织材料的化学处理方法
KR101327630B1 (ko) * 2012-03-05 2013-11-13 울산대학교 산학협력단 아텔로콜라겐을 이용한 췌도세포 이식용 담체의 제조방법 그리고 이를 이용하여 제조된 인공췌장
RU2519219C1 (ru) * 2012-12-25 2014-06-10 Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Московский государственный университет имени М.В. Ломоносова" (МГУ) Биологический перикардиальный протез клапана сердца с хитозановым покрытием и способ его получения
CN106190949B (zh) * 2015-05-08 2020-04-10 上海微创心通医疗科技有限公司 一种干态动物源性胶原纤维组织材料及其制备方法和生物假体
US20200038193A1 (en) * 2017-08-18 2020-02-06 Collagen Matrix, Inc. Collagen-based meniscus implants
EP3852683B1 (en) * 2018-11-01 2024-05-29 Edwards Lifesciences Corporation Transcatheter pulmonic regenerative valve
CN114096567A (zh) * 2019-04-29 2022-02-25 鲍希与洛姆伯股份有限公司 糖磷脂聚合物网络及其用途
AU2020399770A1 (en) * 2019-12-09 2022-01-06 Edwards Lifesciences Corporation Bioprosthetic tissue preparation
US20220195387A1 (en) * 2020-12-21 2022-06-23 The Research Foundation For The State University Of New York Compositions, apparatuses and methods for making and using bioscaffolds
US20220288276A1 (en) * 2021-03-11 2022-09-15 Precise Bio Inc. Artificial endothelial keratoplasty graft and methods of preparation thereof
US20220331488A1 (en) * 2021-04-18 2022-10-20 TissX, Inc. Prosthetic Tissue Treatment For Desirable Mechanical Properties
US20220346926A1 (en) * 2021-04-28 2022-11-03 Medos International Sàrl Systems and methods for using structured tissue augmentation constructs in soft tissue fixation repair
EP4279027A3 (en) * 2022-04-29 2024-02-07 Cortronik GmbH Biological cover

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5782931A (en) * 1996-07-30 1998-07-21 Baxter International Inc. Methods for mitigating calcification and improving durability in glutaraldehyde-fixed bioprostheses and articles manufactured by such methods
CN104334121A (zh) * 2012-06-04 2015-02-04 爱德华兹生命科学公司 预组装的生物假体瓣膜和密封导管
CN105194733A (zh) * 2014-06-30 2015-12-30 王静 一种化学处理人工瓣膜的方法
CN105326581A (zh) * 2015-09-29 2016-02-17 中国科学院金属研究所 一种制备聚乙二醇-蛋白质纤维复合人工心脏瓣膜的方法
CN105963783A (zh) * 2016-04-29 2016-09-28 陕西瑞盛生物科技有限公司 一种生物补片的交联方法以及交联生物补片
CN106913909A (zh) * 2017-02-27 2017-07-04 杭州启明医疗器械有限公司 一种干燥人工生物瓣膜及其制备方法
CN108785747A (zh) * 2018-07-07 2018-11-13 四川大学 一种提高生物材料抗钙化性能的处理方法及其生物材料
CN109260517A (zh) * 2018-09-19 2019-01-25 杭州启明医疗器械有限公司 一种可预装干燥生物心脏瓣膜及其制备方法

Also Published As

Publication number Publication date
US20210260247A1 (en) 2021-08-26
EP3854425A4 (en) 2022-08-31
EP3854425A1 (en) 2021-07-28
EP3854425C0 (en) 2023-07-26
EP3854425B1 (en) 2023-07-26
CN109260517A (zh) 2019-01-25
CN109260517B (zh) 2020-10-30

Similar Documents

Publication Publication Date Title
WO2020057412A1 (zh) 一种可预装干燥生物心脏瓣膜及其制备方法
WO2020057414A1 (zh) 一种可快速吸水展平的干燥生物心脏瓣膜及其制备方法
WO2020057415A1 (zh) 一种可快速复水的预装式生物心脏瓣膜及其制备方法
WO2021239080A1 (zh) 一种兼具抗凝血和抗钙化性能的生物瓣膜及其制备方法
CN111166938B (zh) 一种非戊二醛可预装干燥生物瓣膜材料及制备方法和应用
JP6117250B2 (ja) コラーゲン被覆組織ベース膜及び方法
CN111420120A (zh) 一种具有抗凝血和抗钙化功能的生物瓣膜及其制备方法
WO2013063842A1 (zh) 一种边缘刚性化人工生物瓣膜的制备方法
WO2022143700A1 (zh) 抗折痕的脱水交联生物材料及其制备方法和应用
US20110224779A1 (en) Thin collagen tissue for medical device applications
KR20170128385A (ko) 탈세포화 조직을 사용한 유착 방지재 및 대용 생체막
CN109833519A (zh) 一种人工生物瓣膜的方法
WO2022057841A1 (zh) 一种人工生物心脏瓣膜及其制备方法
WO2022134858A1 (zh) 生物假体组织的处理方法和生物假体心脏瓣膜
CN104028434B (zh) 一种在钛表面构建层粘连蛋白/肝素/SDF-1α抗凝及诱导内皮化多功能层的方法
EP4108265A1 (en) Functionalized biological matrix material, preparation method therefor and use thereof
CN115087470A (zh) 一种功能化生物基质材料及其制备方法和应用
CN116271236A (zh) 一种双键后交联生物瓣膜材料及其制备和应用
EP4135786A1 (en) Method of processing collagen-based tissue for bioprosthetic devices
CN112674908B (zh) 一种耐折弯的干燥生物心脏瓣膜及其制备方法
WO2020038293A1 (zh) 一种采用酶交联和茶多酚组合联用处理生物瓣膜的方法
CN109395164B (zh) 一种干化的动物细胞外基质材料的制备方法
CN109820624A (zh) 一种采用光交联处理生物瓣膜的方法
WO2024103389A1 (zh) 醛基交联后双键聚合制备生物瓣膜材料的方法及生物瓣膜材料和应用
WO2023088330A1 (zh) 一种生物瓣膜材料及其制备方法和应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19862226

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2019862226

Country of ref document: EP

Effective date: 20210419