CN111588909A - Anti-calcification method for biomedical material - Google Patents
Anti-calcification method for biomedical material Download PDFInfo
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- 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
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- 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
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- 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
- A61L27/3629—Intestinal tissue, e.g. small intestinal submucosa
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- 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
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- 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
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- 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
- A61L2400/02—Treatment of implants to prevent calcification or mineralisation in vivo
Abstract
The invention discloses a method for preventing calcification of biomedical materials, which comprises the following steps: s1, treating glutaraldehyde to obtain high-purity monomer glutaraldehyde; s2, adjusting the pH value of the first monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material in the first monomer glutaraldehyde solution, keeping the temperature and keeping the biomedical material for a certain time; s3, adjusting the pH value of the first alcohol solution to 5.0-9.0, soaking the biomedical material obtained in the step S2 in the first alcohol solution, keeping the temperature and keeping the temperature for a certain time; s4, adjusting the pH value of the second monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S3 in the second monomer glutaraldehyde solution, keeping the temperature and keeping the temperature for a certain time; s5, adjusting the pH value of the third monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S4 in the third monomer glutaraldehyde solution, keeping the temperature and continuously storing to obtain the calcification-preventing biomedical material.
Description
Technical Field
The invention relates to the technical field of medical methods, in particular to a method for preventing calcification of a biomedical material.
Background
Biomedical materials are used as a foundation for researching artificial organs and medical instruments, become important branches of modern material disciplines, and particularly become hot spots for research and development by competition among scientists of various countries along with the vigorous development and major breakthrough of biotechnology, and related reports of application and research of related medical materials and medical materials in the field of high and new medical technology are endless in recent years.
The biomedical materials are also called biological materials, the traditional fields mainly comprise artificial organs (orthopedic implants, artificial bones, artificial joints, artificial prostheses and the like) supporting the movement function, artificial organs (artificial blood vessels, artificial heart valves and the like) having the blood circulation function, artificial organs having the plastic and cosmetic functions, artificial organs having the sensory functions (artificial crystals, artificial cochlea and the like) and the like, and the novel fields mainly comprise molecular diagnosis, 3D printing and the like.
Although biomedical materials have been applied with great success, a lot of problems are exposed in long-term clinical application, and the outstanding effects are that the requirements of clinical application cannot be well met in terms of functionality, immunity, service life and the like. For example, the death rate of the heart valve reaches 58 percent after the heart valve is implanted for 12 years, the restenosis rate of the blood vessel reaches about 10 percent after the blood vessel stent is implanted, the validity period of the artificial joint is 12 to 15 years for the old group, and the middle-young group only reaches about 5 years. Among other things, calcification of the material seriously affects the functionality of the material. At present, the principle of the calcification rate of biomedical materials has not been fully studied. However, factors that are thought to affect the rate of calcification include the age of the patient, the presence of metabolic disturbances, dietary factors, the presence of infection, parenteral calcium administration, dehydration, in situ deformation of the bioprosthesis, inadequate anticoagulant therapy during the initial period of surgical implantation, and immune host-tissue response.
In recent years, much effort has been made to prevent calcification of biomedical materials. The technologies that have been harvested from these efforts can be broadly divided into two categories: it relates to the pre-or post-treatment of glutaraldehyde-fixed tissue with one or more compounds that inhibit calcification, as described in the literature: Carpentier-Edwards thermal Process A Method for extracting Calcium Binding Sites from Perical Tissue and involves fixing the Tissue with a compound other than glutaraldehyde, thereby reducing calcification.
The biological valve material is generally tanned and preserved by glutaraldehyde, glutaraldehyde can increase the thermal shrinkage temperature and mechanical tensile strength of tissues and reduce immunogenicity, but free aldehyde groups have a cytotoxic effect and reduce calcium-dependent ATPase activity on cell membranes, so that the calcium content in cells is increased, and tissues are calcified.
Although some of these techniques have proven effective in reducing material calcification, there remains a need in the art for further improvements to the prior art or for the development of new anti-calcification techniques.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for preventing calcification of a biomedical material.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method of preventing calcification of a biomedical material, comprising:
s1, treating glutaraldehyde to obtain high-purity monomer glutaraldehyde;
s2, adjusting the pH value of the first monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material in the first monomer glutaraldehyde solution, keeping the temperature and keeping the biomedical material for a certain time;
s3, adjusting the pH value of the first alcohol solution to 5.0-9.0, soaking the biomedical material obtained in the step S2 in the first alcohol solution, keeping the temperature and keeping the temperature for a certain time;
s4, adjusting the pH value of the second monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S3 in the second monomer glutaraldehyde solution, keeping the temperature and keeping the temperature for a certain time;
s5, adjusting the pH value of the third monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S4 in the third monomer glutaraldehyde solution, keeping the temperature and continuously storing to obtain the calcification-preventing biomedical material.
Further, the step S1 of treating glutaraldehyde specifically includes subjecting glutaraldehyde to gas membrane separation, reduced-pressure fractional distillation, and nanofiltration membrane separation, to remove polymeric substances of glutaraldehyde dimer, glutaraldehyde trimer, and glutaraldehyde tetramer in glutaraldehyde, and obtain high-purity monomeric glutaraldehyde.
Further, the first monomer glutaraldehyde solution in step S2 includes one or more of phosphoric acid buffer solution and glutaraldehyde; the concentration of the first monomer glutaraldehyde solution is 0.1% -2%; the pH of the first monomeric glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the first glutaraldehyde solution is adjusted to 3-50 ℃; the biomedical material is continuously soaked in the first glutaraldehyde solution for 4 hours to 10 days.
Further, the substance of the first alcohol solution in step S3 includes one or more of alcohols, phosphoric acid buffer solution, HEPES buffer solution; the alcohol comprises one or more of ethanol, n-propanol, isopropanol, n-butanol and isobutanol; the volume of the alcohol in the first alcohol solution accounts for 10-100%; the pH of the first alcohol solution is adjusted to 7.1-7.8; the temperature of the first alcohol solution is adjusted to 3-40 ℃; the biomedical material is continuously soaked in the first alcohol solution for 10 hours to 3 days.
Further, the second glutaraldehyde solution in step S4 includes one or more of a phosphate buffer solution and glutaraldehyde; the concentration of the second glutaraldehyde is 0.05% -2%; the pH of the second glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the second glutaraldehyde solution is adjusted to 3-45 ℃; the biomedical material is continuously soaked in the second glutaraldehyde solution for 4 hours to 7 days.
Further, the third glutaraldehyde solution in step S5 includes one or more of a phosphate buffer solution and glutaraldehyde; the concentration of the third glutaraldehyde is 0.1% -1%; the pH of the third glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the third glutaraldehyde solution is adjusted to 3-40 ℃; the biomedical material is continuously soaked in a third glutaraldehyde solution and stored for a long time.
Further, the volume of the alcohol in the first alcohol solution is 60-100%.
Further, the biomedical material is mammalian tissue; the mammal tissue is one or more of pericardial tissue, valve tissue, vascular tissue, adventitia, pleura, peritoneum, achilles tendon, and ligament.
Further, the step S2 and/or the step S3 and/or the step S4 and/or the step S5 are performed under non-oxidizing conditions including keying of an antioxidant, inert gas shielding, dark light for enhancing the performance of the biomedical material.
Further, an antioxidant is added into the first monomer glutaraldehyde solution and/or the first alcohol solution and/or the second monomer glutaraldehyde solution and/or the third monomer glutaraldehyde solution, and the antioxidant comprises ascorbic acid.
Furthermore, one or more of formaldehyde, a surfactant, ethanol and isopropanol are added into the first monomer glutaraldehyde solution and/or the second monomer glutaraldehyde solution and/or the third monomer glutaraldehyde solution so as to improve the practical efficacy of the solution.
Further, one or more of formaldehyde, a surfactant, glutaraldehyde and isopropanol are included in the first alcohol solution to improve the practical efficacy of the solution.
Furthermore, phosphoric acid buffer solution and/or HEPES buffer solution are/is adopted when the first monomer glutaraldehyde solution, the first alcohol solution, the second monomer glutaraldehyde solution and the third monomer glutaraldehyde solution are prepared.
Compared with the prior art, the glutaraldehyde adopted by the invention is high-purity monomer glutaraldehyde which is obtained by removing polymer substances such as glutaraldehyde dimers, glutaraldehyde trimers, glutaraldehyde tetramers and the like through gas membrane separation, reduced-pressure fractional distillation and nanofiltration membrane separation. The biomedical material is cross-linked by monomer glutaraldehyde with a certain concentration, then is processed by alcohol, and is stored in monomer glutaraldehyde solution after sterilization. Thereby obtaining the biomedical material with excellent calcification-preventing effect.
Drawings
Fig. 1 is a flowchart of a method for preventing calcification of a biomedical material according to an embodiment.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict.
The invention aims to provide a method for preventing calcification of biomedical materials, aiming at the defects of the prior art.
Example one
The embodiment provides a method for preventing calcification of a biomedical material, as shown in fig. 1, including:
s11, treating glutaraldehyde to obtain high-purity monomer glutaraldehyde;
s12, adjusting the pH value of the first monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material in the first monomer glutaraldehyde solution, keeping the temperature and keeping the biomedical material for a certain time;
s13, adjusting the pH value of the first alcohol solution to 5.0-9.0, soaking the biomedical material obtained in the step S12 in the first alcohol solution, keeping the temperature and keeping the temperature for a certain time;
s14, adjusting the pH value of the second monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S13 in the second monomer glutaraldehyde solution, keeping the temperature and keeping the temperature for a certain time;
s15, adjusting the pH value of the third monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S14 in the third monomer glutaraldehyde solution, keeping the temperature and continuously storing to obtain the calcification-preventing biomedical material.
In step S11, glutaraldehyde is treated to obtain monomeric glutaraldehyde with high purity.
In this embodiment, the glutaraldehyde is high-purity monomeric glutaraldehyde obtained by removing polymeric substances such as glutaraldehyde dimer, glutaraldehyde trimer, and glutaraldehyde tetramer through gas membrane separation, reduced-pressure fractional distillation, and nanofiltration membrane separation. Adjusting the pH of the first, second and third monomer glutaraldehyde solution to 5.0-9.0, preferably to 5.0-7.0, the glutaraldehyde is unstable under alkaline conditions and readily polymerizes to form polymers. If a solution with a pH of 8 or more is generally inactive within 4 weeks, the activated basic glutaraldehyde should be used for a period of time not exceeding two weeks. The biomedical material is soaked in glutaraldehyde solution and maintained at certain temperature for certain time. The basic structural unit of collagen is procollagen, which is fibrous protein composed of three alpha-peptide chains, twisted into a three-strand helical configuration, with a length of 300nm and a diameter of 1.5 nm. Triple helix means that each type of collagen is formed by three identical or different peptide chains. The triple-helix structure of collagen is completely loosened to form 3 free peptide chains, the relative molecular mass is from thousands to tens of thousands, and the molecular weight distribution is very wide. The cross-linking by adopting the monomer glutaraldehyde has the advantages that the monomer glutaraldehyde only can be cross-linked in the peptide chain, and the cross-linking is firmer; and the glutaraldehyde polymers are crosslinked between the strands of the collagen, so-called pseudo crosslinking is generated, and in the long-term use process of the biomaterial in the later period, the pseudo crosslinking is broken to generate a large amount of free aldehyde groups, so that the crosslinking degree of the biomaterial is greatly reduced.
In step S12, the pH of the first glutaraldehyde monomer solution is adjusted to 5.0-9.0, and the biomedical material is immersed in the first glutaraldehyde monomer solution, maintained at a certain temperature, and kept for a certain time.
In the embodiment, the main substance of the first monomer glutaraldehyde solution comprises one or more of phosphoric acid buffer solution and glutaraldehyde;
the concentration of the first monomer glutaraldehyde solution is 0.1% -2%;
the pH of the first monomeric glutaraldehyde solution is adjusted to 7.1-7.8;
the temperature of the first glutaraldehyde solution is adjusted to 3-50 ℃;
the biomedical material is continuously soaked in the first glutaraldehyde solution for 4 hours to 10 days.
Specifically, when the first glutaraldehyde solution is used, the solution is mixed with a prepared phosphoric acid buffer solution to prepare a 0.5% glutaraldehyde aqueous solution. The pH of the aqueous glutaraldehyde solution was adjusted to 7.4 using 1mol/L hydrochloric acid solution and 1mol/L sodium hydroxide solution. Completely immersing the biomedical material needing anti-calcification treatment in the solution. The glutaraldehyde solution containing the biomedical material was left at 25 ℃ for 70 hours.
In step S13, the pH of the first alcohol solution is adjusted to 5.0 to 9.0, and the biomedical material obtained in step S12 is immersed in the first alcohol solution, maintained at a certain temperature, and kept for a certain period of time.
The main substance of the first alcohol solution comprises one or more of alcohol, phosphoric acid buffer solution and HEPES buffer solution; wherein the alcohol comprises one or more of ethanol, n-propanol, isopropanol, n-butanol, and isobutanol; in addition, the first alcohol solution may contain other chemical components to ensure or enhance the actual efficacy of the solution. Such as formaldehyde, surfactants, glutaraldehyde, isopropanol, and the like.
The volume of the alcohol in the first alcohol solution accounts for 10-100 percent; preferably, the first alcohol solution comprises 60% to 100% alcohol by volume.
The pH of the first alcohol solution is adjusted to 7.1-7.8; the temperature of the first alcohol solution is adjusted to 3-40 ℃;
the biomedical material is continuously soaked in the first alcohol solution for 10 hours to 3 days.
Specifically, anhydrous ethanol and HEPES buffer solution are mixed, the volume of the anhydrous ethanol accounts for 90%, and 1mol/L hydrochloric acid solution and 1mol/L sodium hydroxide solution are used for adjusting the pH value of the glutaraldehyde aqueous solution to 7.4. The biomedical material processed in step S12 is washed clean with physiological saline and then completely immersed in the solution. The glutaraldehyde solution containing the biomedical material is placed in an environment at 25 ℃ for 40 hours.
In step S14, the pH of the second glutaraldehyde monomer solution is adjusted to 5.0-9.0, and the biomedical material obtained in step S13 is soaked in the second glutaraldehyde monomer solution, kept at a certain temperature and kept for a certain time.
The second glutaraldehyde solution comprises one or more of phosphoric acid buffer solution and glutaraldehyde;
the concentration of the second glutaraldehyde is 0.05% -2%;
the pH of the second glutaraldehyde solution is adjusted to 7.1-7.8;
the temperature of the second glutaraldehyde solution is adjusted to 3-45 ℃;
the biomedical material is continuously soaked in the second glutaraldehyde solution for 4 hours to 7 days.
Specifically, a second glutaraldehyde solution, formaldehyde and a phosphoric acid buffer solution are mixed. A0.6% aqueous solution of glutaraldehyde was prepared, and the pH of the aqueous solution of glutaraldehyde was adjusted to 7.4 using 1mol/L hydrochloric acid solution and 1mol/L sodium hydroxide solution. The biomedical material of step S13 is washed clean with physiological saline and then completely immersed in the solution. The glutaraldehyde solution containing the biomedical material was left at 25 ℃ for 70 hours.
In step S15, the pH of the third monomer glutaraldehyde solution is adjusted to 5.0-9.0, and the biomedical material obtained in step S14 is soaked in the third monomer glutaraldehyde solution, kept at a certain temperature and kept for a while to obtain the calcification-preventing biomedical material.
The third glutaraldehyde solution comprises one or more of phosphoric acid buffer solution and glutaraldehyde;
the concentration of the third glutaraldehyde is 0.1% -1%;
the pH of the third glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the third glutaraldehyde solution is adjusted to 3-40 ℃;
the biomedical material is continuously soaked in the third glutaraldehyde solution and stored for a long time.
Specifically, a third glutaraldehyde solution and a phosphoric acid buffer solution are mixed. 0.4% glutaraldehyde solution in water was prepared. The pH of the aqueous glutaraldehyde solution was adjusted to 7.4 using 1mol/L hydrochloric acid solution and 1mol/L sodium hydroxide solution. The biomedical material processed in step S14 is washed clean with physiological saline and then completely immersed in the solution. The glutaraldehyde solution with the biomedical material is stored in an environment with the temperature of 25 ℃ for a long time.
In this embodiment, an antioxidant, preferably ascorbic acid, is added to the first monomeric glutaraldehyde solution and/or the first alcohol solution and/or the second monomeric glutaraldehyde solution and/or the third monomeric glutaraldehyde solution.
Each of the glutaraldehyde solutions mentioned in this example may be used as the final disinfecting or sterilizing solution. In addition, the first monomer glutaraldehyde solution and/or the second monomer glutaraldehyde solution and/or the third monomer glutaraldehyde solution can be added with other chemical components possibly to ensure or improve the actual efficacy of the solution. Such as formaldehyde, surfactants, ethanol, isopropanol, and the like.
Other chemical components may be included in the first alcohol solution to ensure or enhance the actual efficacy of the solution. Such as formaldehyde, surfactants, glutaraldehyde, isopropanol, and the like.
In practice, terminal sterilization is preferably performed in sealed containers in which the biomedical material will be stored and transported until implantation is complete. In this process, the storage temperature requirements must be met, with a temperature range of 3 ℃ to 40 ℃ generally being preferred. The pH of the preservation solution is preferably 7.1 to 7.8, more preferably 7.4. The pH value is kept to be stable at a certain temperature. The containers are transported, placed in a hospital or other location, and stored until use.
The present embodiment contemplates that step S12 and/or step S3 and/or step S4 and/or step S15 are performed under non-oxidizing conditions, wherein the non-oxidizing conditions include keying in of antioxidants, inert gas shielding, dim light, for enhancing the performance of the biomedical material.
Phosphoric acid buffer solution and/or HEPES buffer solution were used for preparing the glutaraldehyde solution and the alcohol solution.
Each of the steps (steps S12-S15) is preferably performed in as sterile an environment as possible, such as a ten-thousand class clean room, a hundred class laminar flow chamber, a biosafety cabinet, etc., to reduce bioburden and particulate matter in the environment, etc.
The biomedical material useful in this embodiment is preferably mammalian tissue, particularly pericardial tissue, valve tissue, vascular tissue, adventitia, pleura, peritoneum, achilles tendon, ligament, and the like.
In a preferred embodiment, the method of anti-calcification of biomedical materials is applied to any component containing mammalian tissue. Various implantable articles such as heart valves, valved conduits, vascular prostheses, biological patches, ligaments, and the like, are used in replacement or repair procedures.
Compared with the prior art, the glutaraldehyde adopted in the embodiment is high-purity monomeric glutaraldehyde obtained by removing polymer substances such as glutaraldehyde dimers, glutaraldehyde trimers and glutaraldehyde tetramers through gas membrane separation, reduced-pressure fractional distillation and nanofiltration membrane separation. The biomedical material is cross-linked by monomer glutaraldehyde with a certain concentration, then is processed by alcohol, and is stored in monomer glutaraldehyde solution after sterilization. Thereby obtaining the biomedical material with excellent calcification-preventing effect.
Example two
The present embodiment provides a method for preventing calcification of a biomedical material, which is different from the first embodiment in that:
this example illustrates the following three experiments comparing the difference between the process of the present invention and the conventional treatment process.
Experiment one:
the cross-linking degree test and inspection method is usually a leather hot-shrinking method, and the cross-linking degree of the collagen tissue is positively correlated with the shrinking temperature of the heated collagen tissue. Crosslinking degree comparison experiment:
the high-purity monomeric glutaraldehyde obtained by removing polymer substances such as glutaraldehyde dimers, glutaraldehyde trimers and glutaraldehyde tetramers through gas membrane separation, reduced-pressure fractional distillation and nanofiltration membrane separation is taken as a first group (A1, A2, A3, A4 and A5), and the glutaraldehyde which is not subjected to the above-mentioned process treatment is taken as a second group (B1, B2, B3, B4 and B5). Bovine pericardium was treated separately. After the product is prepared, the product is put into a fatigue testing system to accelerate fatigue for one hundred million times, and then the sample thermal shrinkage temperature is tested by a leather thermal shrinkage method.
The method comprises cutting each group of test pieces into 10cm × 50cm, washing with distilled water, inserting the test pieces into upper and lower hooks of leather thermometer, attaching 3g weight to the other end, heating with distilled water as medium from room temperature, raising temperature by 2 deg.C per minute, and using the shrinkage pointer of the test pieces as its thermal shrinkage temperature. In this example, the data of the cross-linking degree test after the fatigue test are shown in Table 1:
TABLE 1
Experiment two:
the content of free aldehyde groups in the liquid after the fatigue test.
The solutions of the first group (A1, A2, A3, A4, A5) and the second group (B1, B2, B3, B4, B5) in the first experiment are taken for testing, and the content of free aldehyde groups is tested, as shown in Table 2.
Two methods are commonly used for the detection of aldehyde groups, one is a newly prepared silver ammonia solution, and the other is a newly prepared copper hydroxide suspension, and the two methods are respectively described below.
(one) testing with freshly prepared silver ammonia solution
1: preparation of silver ammonia solution
As the experiment needs to use a newly prepared silver-ammonia solution, the silver-ammonia solution needs to be prepared at present, and the method for preparing the silver-ammonia solution comprises the following steps: and dropwise adding ammonia water into the silver nitrate solution, and stopping dropwise immediately until the white precipitate generated at the beginning is just completely dissolved, so that the silver ammonia solution is prepared. The white precipitate produced in the intermediate process is silver hydroxide and the equation for the reaction is as follows:
2: detection of aldehyde groups in organic substances by silver ammonia solution
Taking a clean test tube, injecting an organic solution to be detected into the test tube, adding a silver ammonia solution, uniformly mixing, putting the test tube into a water bath for heating, controlling the temperature of the water bath to be 60-70 ℃, and if a bright silver mirror appears in the test tube finally, proving that the organic molecule contains aldehyde groups, if acetaldehyde, the related reaction is as follows: if formaldehyde, the equation is as follows:
although formaldehyde is a monoaldehyde, the structure is quite special and corresponds to a dialdehyde, so that the consumption amount of the silver ammonia solution is twice of that of other monoaldehydes in the amount of the substances. It is for this reason that it is usually necessary to verify whether the monoaldehyde is formaldehyde or not in the case of xiti in which the monoaldehyde reacts with a silver-ammonia solution
The method is used for detecting aldehyde groups, and the experimental key points are as follows: the ammonia water cannot be excessive when preparing the silver-ammonia solution, and a water bath is required to be heated.
Silver diammine hydroxide is the solute in silver ammonia solution, and is a strong electrolyte and a strong base. Because bright silver mirror is produced in the experimental process, the reaction is also called silver mirror reaction chemically. After the test, the silver on the inner wall of the test tube can be removed by dilute sulfuric acid.
(II) examination with freshly prepared suspension of copper hydroxide
1: preparation of copper hydroxide suspension
The method for detecting aldehyde group also needs to be used for preparing the copper hydroxide suspension on site, and the preparation method comprises the following steps: a small amount of copper sulfate solution was added dropwise to the sodium hydroxide solution until suspension appeared, thereby preparing a copper hydroxide suspension. The reaction equation in this process is:
2: detection of aldehyde groups in organic materials with freshly prepared copper hydroxide suspension
Taking a clean test tube, injecting an organic solution to be detected into the test tube, adding a newly prepared copper hydroxide suspension, uniformly mixing, directly placing the test tube on an alcohol lamp, heating to boil, and if brick red precipitate finally appears, indicating that the molecular of the tested organic substance contains aldehyde group, wherein the brick red precipitate is cuprous oxide.
If the organic substance to be examined is acetaldehyde, the relevant reactions are:
the method is used for detecting aldehyde groups, and the experimental key points are as follows: when preparing a freshly prepared copper hydroxide suspension, an excess of sodium hydroxide is required, so that a small amount of copper sulfate solution is required for the preparation. In addition, when the aldehyde group is detected, the aldehyde group needs to be directly heated to boiling, the boiling time cannot be too long, and the decomposition of copper hydroxide is prevented to generate black copper oxide.
The principle of the experiment can be used for the detection of diabetes in medicine, because the urine of the diabetic contains glucose, and the glucose molecule contains aldehyde group.
And (3) knotting: the test of aldehyde group is carried out under alkaline condition. Common aldehyde group-containing substances are: aldehydes, formic acid, formate, glucose, maltose, etc., all of which can react with silver mirror and also react with newly prepared copper hydroxide suspension to generate brick red precipitate. In addition, some saccharides do not have an aldehyde group per se, but can be hydrolyzed under acidic conditions to form a substance having an aldehyde group. For example, sucrose does not have an aldehyde group in the molecule, but glucose, which is one of the hydrolysis products, contains an aldehyde group. Both starch and cellulose hydrolysates are glucose. In the case of testing the hydrolysate of glucide, since the hydrolysis is carried out under acidic conditions, the hydrolysate is acidic and needs to be adjusted to be alkaline for testing.
The content of free aldehyde groups in the liquid after fatigue testing is shown in table 2:
TABLE 2
Experiment three:
the high-purity monomeric glutaraldehyde obtained by removing polymer substances such as glutaraldehyde dimers, glutaraldehyde trimers and glutaraldehyde tetramers through gas membrane separation, reduced-pressure fractional distillation and nanofiltration membrane separation is taken as a first group (C1, C2, C3, C4, C5, C6, C7 and C8), and the glutaraldehyde which is not treated by the process is taken as a second group (D1, D2, D3, D4, D5, D6, D7 and D8). Bovine pericardium was treated separately. The bovine pericardium obtained was cut into 10mm × 10mm samples, and rinsed with sterile physiological saline. Under aseptic condition, the bovine pericardium is respectively implanted under the back skin of a mouse, the wound is sewed by using 5-0Prolene wires, the bovine pericardium is sent to an animal laboratory for routine breeding, and the bovine pericardium is subjected to routine penicillin injection for 3d (20 ten thousand U/(kg. d)) after operation.
Tissue calcium content detection: and (3) overnarcotizing rats (178-327 g) with pentobarbital sodium after the bovine pericardium test piece is implanted for 60 days, taking out each group of test pieces, and detecting the calcium content of the tissues: rinsing each group of test pieces with distilled water for 3 times and 10 min/time, drying surface water by filter paper, drying at 80 deg.C, and measuring calcium content by atomic absorption flame method. The results are shown in Table 3:
TABLE 3
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (13)
1. A method of preventing calcification of a biomedical material, comprising:
s1, treating glutaraldehyde to obtain high-purity monomer glutaraldehyde;
s2, adjusting the pH value of the first monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material in the first monomer glutaraldehyde solution, keeping the temperature and keeping the biomedical material for a certain time;
s3, adjusting the pH value of the first alcohol solution to 5.0-9.0, soaking the biomedical material obtained in the step S2 in the first alcohol solution, keeping the temperature and keeping the temperature for a certain time;
s4, adjusting the pH value of the second monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S3 in the second monomer glutaraldehyde solution, keeping the temperature and keeping the temperature for a certain time;
s5, adjusting the pH value of the third monomer glutaraldehyde solution to 5.0-9.0, soaking the biomedical material obtained in the step S4 in the third monomer glutaraldehyde solution, keeping the temperature and continuously storing to obtain the calcification-preventing biomedical material.
2. The method for preventing calcification of a biomedical material as claimed in claim 1, wherein the step S1 of treating glutaraldehyde specifically includes subjecting glutaraldehyde to gas membrane separation, reduced-pressure fractional distillation, and nanofiltration membrane separation, and removing polymeric substances of glutaraldehyde dimer, glutaraldehyde trimer, and glutaraldehyde tetramer in glutaraldehyde to obtain high-purity monomeric glutaraldehyde.
3. The method for preventing calcification of a biomedical material as claimed in claim 1, wherein said first monomeric glutaraldehyde solution in step S2 includes one or more of phosphoric acid buffer solution and glutaraldehyde; the concentration of the first monomer glutaraldehyde solution is 0.1% -2%; the pH of the first monomeric glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the first glutaraldehyde solution is adjusted to 3-50 ℃; the biomedical material is continuously soaked in the first glutaraldehyde solution for 4 hours to 10 days.
4. The method for preventing calcification of a biomedical material as claimed in claim 3, wherein the substance of the first alcohol solution in step S3 includes one or more of alcohol, phosphoric acid buffer solution, HEPES buffer solution; the alcohol comprises one or more of ethanol, n-propanol, isopropanol, n-butanol and isobutanol; the volume of the alcohol in the first alcohol solution accounts for 10-100%; the pH of the first alcohol solution is adjusted to 7.1-7.8; the temperature of the first alcohol solution is adjusted to 3-40 ℃; the biomedical material is continuously soaked in the first alcohol solution for 10 hours to 3 days.
5. The method for preventing calcification of a biomedical material as claimed in claim 4, wherein said second glutaraldehyde solution in step S4 includes one or more of phosphate buffer solution and glutaraldehyde; the concentration of the second glutaraldehyde is 0.05% -2%; the pH of the second glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the second glutaraldehyde solution is adjusted to 3-45 ℃; the biomedical material is continuously soaked in the second glutaraldehyde solution for 4 hours to 7 days.
6. The method for preventing calcification of a biomedical material as claimed in claim 5, wherein the substance of the third glutaraldehyde solution in step S5 includes one or more of phosphate buffer solution and glutaraldehyde; the concentration of the third glutaraldehyde is 0.1% -1%; the pH of the third glutaraldehyde solution is adjusted to 7.1-7.8; the temperature of the third glutaraldehyde solution is adjusted to 3-40 ℃; the biomedical material is continuously soaked in a third glutaraldehyde solution and stored for a long time.
7. The method of claim 6, wherein the first alcohol solution comprises 60% to 100% alcohol by volume.
8. The method of claim 1, wherein the biomedical material is mammalian tissue; the mammal tissue is one or more of pericardial tissue, valve tissue, vascular tissue, adventitia, pleura, peritoneum, achilles tendon, and ligament.
9. The method for preventing calcification of a biomedical material as claimed in claim 1, wherein said step S2 and/or step S3 and/or step S4 and/or step S5 are performed under non-oxidizing conditions, said non-oxidizing conditions including keying of antioxidants, inert gas shielding, dim light, for enhancing the performance of the biomedical material.
10. The method for preventing calcification of a biomedical material as claimed in claim 1, wherein an antioxidant is added to said first monomeric glutaraldehyde solution and/or said first alcohol solution and/or said second monomeric glutaraldehyde solution and/or said third monomeric glutaraldehyde solution, said antioxidant comprising ascorbic acid.
11. The method for preventing calcification of a biomedical material as claimed in claim 10, wherein one or more of formaldehyde, surfactant, ethanol and isopropanol are added to the first monomeric glutaraldehyde solution and/or the second monomeric glutaraldehyde solution and/or the third monomeric glutaraldehyde solution to improve the actual efficacy of the solution.
12. The method of claim 1, wherein the first alcohol solution comprises one or more of formaldehyde, a surfactant, glutaraldehyde, and isopropanol to improve the effective efficacy of the solution.
13. The method for preventing calcification of a biomedical material as claimed in claim 1, wherein a phosphoric acid buffer solution and/or a HEPES buffer solution is/are used in the preparation of the first glutaraldehyde solution, the first alcohol solution, the second glutaraldehyde solution and the third glutaraldehyde solution.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1136780A (en) * | 1993-10-21 | 1996-11-27 | 密执安大学 | Method for making calcification-resistant bioprosthetic tissue |
| US6214055B1 (en) * | 1998-10-30 | 2001-04-10 | Mures Cardiovascular Research, Inc. | Method and kit for rapid preparation of autologous tissue medical devices |
| CN1371750A (en) * | 2002-02-28 | 2002-10-02 | 中南大学湘雅二医院 | Biological cardiac valves 2,3-butanediol calcification prevention modifying method |
| US6561970B1 (en) * | 1994-07-29 | 2003-05-13 | Edwards Lifesciences Corporation | Methods for treating implantable biological tissues to mitigate the calcification thereof and bioprosthetic articles treated by such methods |
| CN101184516A (en) * | 2005-03-25 | 2008-05-21 | 爱德华兹生命科学公司 | Treatment of bioprosthetic tissues to mitigate post implantation calcification |
| CN109589452A (en) * | 2018-11-21 | 2019-04-09 | 杭州嘉和众邦生物科技有限公司 | A kind of anticalcium processing method of biomaterial |
-
2020
- 2020-02-24 CN CN202010111152.4A patent/CN111588909A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1136780A (en) * | 1993-10-21 | 1996-11-27 | 密执安大学 | Method for making calcification-resistant bioprosthetic tissue |
| US6561970B1 (en) * | 1994-07-29 | 2003-05-13 | Edwards Lifesciences Corporation | Methods for treating implantable biological tissues to mitigate the calcification thereof and bioprosthetic articles treated by such methods |
| US6214055B1 (en) * | 1998-10-30 | 2001-04-10 | Mures Cardiovascular Research, Inc. | Method and kit for rapid preparation of autologous tissue medical devices |
| CN1371750A (en) * | 2002-02-28 | 2002-10-02 | 中南大学湘雅二医院 | Biological cardiac valves 2,3-butanediol calcification prevention modifying method |
| CN101184516A (en) * | 2005-03-25 | 2008-05-21 | 爱德华兹生命科学公司 | Treatment of bioprosthetic tissues to mitigate post implantation calcification |
| CN109589452A (en) * | 2018-11-21 | 2019-04-09 | 杭州嘉和众邦生物科技有限公司 | A kind of anticalcium processing method of biomaterial |
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