WO2023003209A1 - Matériau de greffe osseuse comprenant un support dérivé d'os déminéralisé introduit et son procédé de préparation - Google Patents

Matériau de greffe osseuse comprenant un support dérivé d'os déminéralisé introduit et son procédé de préparation Download PDF

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WO2023003209A1
WO2023003209A1 PCT/KR2022/009439 KR2022009439W WO2023003209A1 WO 2023003209 A1 WO2023003209 A1 WO 2023003209A1 KR 2022009439 W KR2022009439 W KR 2022009439W WO 2023003209 A1 WO2023003209 A1 WO 2023003209A1
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bone
demineralized bone
graft material
demineralized
powder
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PCT/KR2022/009439
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English (en)
Korean (ko)
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정홍희
이유진
이재열
이대형
최예지
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한스바이오메드 주식회사
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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
    • A61L27/3608—Bone, e.g. demineralised bone matrix [DBM], bone powder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/28—Bones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
    • A61L27/3645—Connective tissue
    • A61L27/365—Bones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—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
    • A61L27/3691—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 physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
    • 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
    • A61L27/56—Porous materials, e.g. foams or sponges
    • 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
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

Definitions

  • the present invention relates to a bone graft material into which a demineralized bone-derived carrier is introduced and a manufacturing method thereof. More specifically, it relates to a bone graft material containing a demineralized bone-derived carrier, which is composed of only demineralized bone and has excellent shape retention, and a manufacturing method thereof.
  • Naturally occurring bone is composed of organic and inorganic materials.
  • Organic substances include growth factors, cartilage tissue, collagen, and other proteins, and inorganic substances include calcium phosphate.
  • Bone implants are used to augment the natural regeneration process when bone defects or injuries occur, and must be biocompatible.
  • an ideal bone graft should be capable of bone formation, i.e. bone conduction and bone induction simultaneously.
  • Bone transplantation for the purpose of treatment is performed for the purpose of reconstruction of lost bone and the purpose of reconstruction of joint surfaces that secrete synovial fluid. For example, treatment of nonunion fractures, joint fixation, filling of bone cavity, replacement of lost bone and joint parts, strengthening of acetabulum and skull, fusion of growth plate cartilage, bone and muscle transplantation for ligament loss, etc. Bone grafting is being done in the field. Bone reconstruction through bone transplantation includes autologous transplantation in which a part of one's own bone is harvested and transplanted to a target body part, allograft using the bone of someone other than the patient himself, and xenograft using the bone of another animal. there is.
  • demineralized bone is a transplant material that is safe from the risk of disease transmission from the donor site and has excellent bone regeneration ability, so it can be used in orthopedics and dentistry.
  • Bone morphogenetic protein (BMP) present in demineralized bone is a representative material that increases bone regeneration and growth.
  • demineralized bone since it is difficult to maintain the shape of demineralized bone when transplanted alone, it was necessary to link with other materials. Accordingly, graft materials formulated by mixing demineralized bone with glycerol, a chemical carrier, or poloxamer, an organic synthetic polymer, have been developed.
  • the graft material mixed with glycerol has a problem in that the content of demineralized bone is significantly lowered and the bone regeneration action is slow. There was a problem that the burden could go.
  • Patent Document 1 a method of kneading demineralized bone powder with sterilized purified water and molding it into a stick shape has also been devised. There was a problem.
  • An object of the present invention is to provide a bone graft material composed of only demineralized bone and excellent in bone induction and biocompatibility, and excellent in shape retention, and a manufacturing method thereof.
  • another object of the present invention is to provide a demineralized bone-derived carrier and a method for manufacturing the same, which can be used to manufacture a bone graft material composed of only demineralized bone and having excellent bone induction and biocompatibility and excellent shape retention will be.
  • the inventors of the present invention conducted intensive research for the purpose of advantageously solving the above problems.
  • the demineralized bone powder and water are mixed in a predetermined ratio to prepare a demineralized bone paste, and then the demineralized bone paste is treated with an autoclave.
  • demineralized bone powder can be suitably dissolved in water to obtain a demineralized bone-derived carrier having a viscosity suitable for use as a carrier for producing a bone graft material, thereby completing the present invention.
  • the step (a) of preparing demineralized bone powder by demineralizing bone powder
  • step (b) preparing a demineralized bone paste by mixing the demineralized bone powder prepared in step (a) with water;
  • a method for producing a demineralized bone-derived carrier for producing a bone graft material comprising the step (c) of treating the demineralized bone dough prepared in step (b) with an autoclave,
  • a demineralized bone-derived carrier for manufacturing a bone graft material prepared by dissolving demineralized bone in water and having a viscosity of 495 Pa ⁇ s or more and 2000 Pa ⁇ s or less is provided.
  • a method for producing a bone graft material in which the above-described demineralized bone-derived carrier for producing a bone graft material and demineralized bone powder are mixed and freeze-dried.
  • a bone graft material into which the demineralized bone-derived carrier for manufacturing the above-described bone graft material is introduced is provided.
  • a demineralized bone-derived carrier and a method for manufacturing the same, which can be used to manufacture a bone graft material composed of only demineralized bone and having excellent bone induction ability and biocompatibility as well as excellent shape retention.
  • a bone graft material composed of only demineralized bone and having excellent bone induction ability and biocompatibility as well as shape retention and a manufacturing method thereof is provided.
  • FIG. 1 is a photograph of a bone graft material manufactured by the manufacturing method of the present invention.
  • Fig. 2 is a schematic view of the bone graft material shown in Fig. 1;
  • FIG. 3 is a photograph showing a state after molding the mixture for forming a demineralized bone-derived carrier and a bone graft material prepared in Examples 1, 4, and 6 of the present invention with a mold.
  • FIG. 4 is a photograph showing a state after molding the mixture for forming a demineralized bone-derived carrier and a bone graft material prepared in Comparative Examples 1 and 3 of the present invention with a mold.
  • FIG. 5 is a photograph showing a state in which demineralized bone and water are mixed in preparation of a demineralized bone-derived carrier in Comparative Examples 23 and 24 of the present invention.
  • the demineralized bone-derived carrier according to the present invention is used to manufacture a bone graft material composed only of demineralized bone, and is manufactured by a manufacturing method including the following.
  • Step (a) preparing demineralized bone powder by demineralizing bone powder; (b) preparing a demineralized bone paste by mixing the demineralized bone powder prepared in step (a) with water in a specific ratio; Step (c) of treating the demineralized bone dough with an autoclave.
  • Step (a) preparing demineralized bone powder by demineralizing bone powder; (b) preparing a demineralized bone paste by mixing the demineralized bone powder prepared in step (a) with water in a specific ratio; Step (c) of treating the demineralized bone dough with an autoclave.
  • Step (a) of preparing demineralized bone powder by demineralizing bone powder
  • step (a) bone powder is demineralized under specific conditions to produce demineralized bone powder.
  • step (a) may include the following.
  • the bone powder is obtained by pre-processing and pulverizing bone tissue collected from a donor. A method for producing bone powder will be described later.
  • Demineralization is to facilitate the release of bone-inducing proteins such as BMP by removing calcium phosphate present in bone tissue.
  • demineralization is performed using an acidic solution such as HCl, EDTA, formic acid, citric acid, acetic acid, nitric acid, or nitrous acid. can Among these, it is preferable to demineralize using an HCl aqueous solution.
  • the concentration of the acidic solution used at the time of demineralization is 0.3N or more and 1.5N or less.
  • the demineralization time is preferably 2 hours or more, more preferably 2.5 hours or more, preferably 7 hours or less, and 6.5 hours or less, when the concentration of the acidic solution used is 0.3N or more and 0.8N or less.
  • the demineralization time is preferably 0.1 hour or more, more preferably 0.3 hour or more, still more preferably 0.5 hour or more, and 7 hours or less, when the concentration of the acidic solution used is more than 0.8N and 1.5N or less. It is preferable that it is 6.5 hours or less, and it is more preferable that it is 3 hours or less.
  • the temperature at the time of demineralization is not particularly limited, but can be carried out under conditions of, for example, 15°C or more and 25°C or less.
  • the demineralized bone obtained through the demineralization step is neutralized.
  • a neutralization method for example, sodium hydroxide (NaOH) can be used, and also washing several times with phosphate buffer saline, commercially available physiological saline, 0.9% sodium chloride solution, sterile distilled water, etc. or demineralization reaction It can be neutralized by adding it to an acidic solution.
  • the demineralized bone that has undergone the neutralization step is freeze-dried.
  • the freeze-drying method is not limited thereto, but for example, the demineralized bone is spread evenly on a square dish, the square dish is packaged with an air permeable pouch, and then put in a freeze dryer and freeze-dried for 36 hours or more to obtain demineralized bone.
  • a method may be used to achieve a moisture content of less than 6%.
  • the demineralized bone obtained through the freeze-drying step may be sterilized and stored before being used in the next step.
  • sterilization E.O. It can be performed by gas sterilization, gamma ray sterilization, E-beam sterilization, etc.
  • the bone powder used in step (a) of the present invention is prepared by a manufacturing method comprising the following.
  • Pre-processing the bone collected from the donor crushing and sorting.
  • the step of pre-processing the bone collected from the donor includes the following.
  • first delipidation and immunoantigen step removing and washing soft tissue from bone collected from a donor; immersing the washed bones in an organic solvent and stirring to firstly remove lipids and immune antigens (hereinafter sometimes referred to as “first delipidation and immunoantigen step”); Crushing and sorting the bones from which lipids and immune antigens have been removed.
  • Bone tissue is collected and transported from the donor.
  • the bone (bone) suitable for producing the demineralized bone powder used in the present invention autologous, allogeneic, or xenogeneic bone may be used.
  • Useful sources of xenograft may be pigs, horses, or cattle.
  • the bone may be cortical, reticular, or reticular cortical.
  • Preferred bones are cortical allograft bones such as the femur, tibia, radius, and ulna.
  • bone marrow and blood inside the bone are removed, preferably using high-pressure sterilized purified water and a centrifugal separator to remove and wash the bone marrow and blood.
  • the bone tissue Before or after performing the step of removing and washing the soft tissue, after treating the bone tissue with antibiotics, it is preferable to store it in a freezer at -40 ° C or less. Thereby, cells in the bone tissue capable of causing immune rejection can be inactivated.
  • the bone tissue may be frozen and then thawed using a low-speed freezing system.
  • the slow freezing system can be programmed to descend at a rate of 1°C/min using liquid nitrogen, allowing freezing without forming crystals in the tissue. For example, starting at room temperature 4 ° C, dropping at a rate of 1 ° C / min to -4 ° C, dropping from -4 ° C to 20 ° C / min at a rate of -45 ° C, Rising at a rate of 15°C/min to -10°C, descending at a rate of 0.5°C/min at -10°C to -20°C, and descending at a rate of 1°C/min at -20°C to -80°C It is possible to inactivate cells in bone tissue without generating crystals in the tissue by performing an operation to drop the bone to 10°C. In the case of storage thereafter, it is stored in a cryogenic freezer (-70 ° C), and in case of use, it can be melted at 4
  • lipids and immune antigens are removed from bone tissue using various organic solvents such as hot water or chloroform/methanol, 95% ethanol, 70% ethanol, and diethyl ether.
  • organic solvents such as hot water or chloroform/methanol, 95% ethanol, 70% ethanol, and diethyl ether.
  • it is not limited thereto, but, for example, using 5 ml or more and 30 ml or less of diethyl ether per 1 g of bone tissue, at a temperature of 15 ° C. or more and 25 ° C. or less and in a vacuum, for about 3 hours or more and 24 hours or less. After stirring for 2 hours, it may be dried for 2 hours or more and 24 hours or less.
  • the bone tissue after the first step of delipidation and deimmunization is pulverized into various sizes.
  • the crushed bone pieces are separated and sorted according to size using sieves of various sizes.
  • the term bone fragments as used herein refers to relatively small bone fragments such as regular or irregular fibers, chips, splinters, powders, and the like.
  • the grinding and sorting step is not limited to being performed after the first delipidation and deimmunization step, and may optionally be further performed before or after other steps in order to make the particle size of the demineralized bone powder within a desired range.
  • a second delipidation and deimmunization step may be performed to remove lipids and immune antigens that have not yet been removed in the first delipidation and deimmunization step.
  • the type and amount of the solvent used, the stirring time and temperature, and the drying time may be the same as those in the first delipidation and deimmunization step.
  • a demineralized bone paste is prepared by mixing the demineralized bone powder obtained in step (a) with water in a predetermined ratio.
  • demineralized bone dough means a state in which demineralized bone powder and water are mixed, and the mixing method is not particularly limited.
  • the mixing ratio of demineralized bone powder and water is within the above range, the demineralized bone powder is suitably dissolved in water when treated with an autoclave, even though a sufficient amount of demineralized bone is ensured in the demineralized bone-derived carrier obtained, for use in producing a bone graft material. Since it has a viscosity suitable for use as a carrier, a bone graft material manufactured using the resulting demineralized bone-derived carrier can have both excellent bone induction ability and shape retention ability.
  • the particle size of the demineralized bone powder used in step (b) is preferably 500 ⁇ m or less, more preferably 400 ⁇ m or less, still more preferably 300 ⁇ m or less, and most preferably 200 ⁇ m or less.
  • the demineralized bone powder dissolves more favorably in water when treated with an autoclave and has a viscosity suitable for use as a carrier for producing a bone graft material. The durability and compressive strength of the bone graft material manufactured using the carrier can be improved.
  • the water used in step (b) is not particularly limited, but for example, distilled water (DW), sterilized purified water, and the like can be used.
  • DW distilled water
  • sterilized purified water and the like can be used.
  • a demineralized bone-derived carrier having a viscosity suitable for use as a carrier for preparing a bone graft material is prepared by treating the demineralized bone dough prepared in step (b) with an autoclave.
  • the autoclave treatment is performed under conditions of heating and pressurization, and may be performed at a temperature of 100° C. or more and 130° C. or less and a pressure of 1.9 kgf/cm 2 or more and 2.2 kgf/cm 2 or less.
  • the autoclave treatment time can be set according to the demineralization conditions in step (a) (demineralization step).
  • the time of the autoclave treatment is more than 10 minutes and 65 minutes or less under the condition of demineralization for 2 hours or more when the concentration of the acidic solution used in (desalination step) in step (a) is 0.3 N or more and 0.8 N or less.
  • the autoclave treatment is performed during the autoclave treatment, and it is more preferable that the autoclave treatment is performed for a time of 15 minutes or more and 60 minutes or less.
  • concentration of the acidic solution used in (desalination step) in step (a) is greater than 0.8N and less than or equal to 1.5N, autoclave treatment for more than 10 minutes and less than 65 minutes under the condition of demineralization for 0.1 hour or more It is preferable, and autoclave treatment is more preferable for a time of 15 minutes or more and 60 minutes or less.
  • the demineralized bone powder is suitably dissolved in water and has a viscosity suitable for use as a carrier for producing a bone graft material, while minimizing damage to the bone structure to obtain the resulting demineralized bone-derived carrier
  • the durability and compressive strength of the bone graft material manufactured using the material can be improved.
  • the demineralized bone-derived carrier according to the present invention is produced by the above-described method, and since demineralized bone is suitably dissolved in water, it has a viscosity suitable for use as a carrier for producing a bone graft material.
  • the demineralized bone-derived carrier according to the present invention has a viscosity of 495 Pa ⁇ s or more and 2,000 Pa ⁇ s or less, preferably 700 Pa ⁇ s or more and 1500 Pa ⁇ s or less. Since the demineralized bone-derived carrier according to the present invention has such a predetermined viscosity, it can act to enable molding when manufacturing a bone graft material, so that even if the obtained bone graft material is composed of only demineralized bone, it has excellent durability and compressive strength. It is possible to make the bone graft material excellent in shape retention.
  • the method for measuring the viscosity of the demineralized bone-derived carrier according to the present invention is not particularly limited, and can be measured at 1/s shear rate at 25° C., for example, using a rheometer.
  • the bone graft material when a bone graft material is manufactured using the demineralized bone-derived carrier according to the present invention, since the bone graft material can be composed only of demineralized bone, it can have excellent bone inducing ability and biocompatibility.
  • the bone graft material according to the present invention is manufactured by a manufacturing method including the following.
  • step (d) the above-mentioned demineralized bone-derived carrier for preparing a bone graft material and demineralized bone powder are mixed.
  • the demineralized bone powder mixed in step (d) is not limited by its manufacturing method, and may be, for example, produced by the above-described step (a) of the present invention.
  • the demineralized bone powder mixed in step (d) may have a particle size different from that of the demineralized bone powder prepared in step (a) by appropriately adjusting the grinding and sorting conditions in step (a). there is.
  • the particle size of the demineralized bone powder mixed in step (d) is preferably greater than 150 ⁇ m and less than or equal to 2,000 ⁇ m, more preferably greater than or equal to 170 ⁇ m and less than or equal to 1500 ⁇ m, and still more preferably greater than or equal to 200 ⁇ m and less than or equal to 850 ⁇ m.
  • the particle size of the demineralized bone powder mixed in step (d) is within the above range, it can be uniformly mixed with the demineralized bone-derived carrier for preparing the bone graft material, and thus the finally obtained bone graft material can have excellent durability and compressive strength.
  • the mixing ratio of the demineralized bone-derived carrier for bone graft material manufacturing and the demineralized bone powder is within the above range, the demineralized bone-derived bone-derived carrier for bone graft material manufacturing and the demineralized bone powder are uniformly mixed, resulting in excellent durability and compressive strength of the finally obtained bone graft material. can do.
  • step (e) the mixture for forming a bone graft material prepared in step (d) is freeze-dried to prepare a bone graft material composed of demineralized bone.
  • a bone graft material composed of demineralized bone is manufactured by molding a mixture for forming a bone graft material into a desired shape using a mold and freeze-drying it.
  • the freeze-drying method is not particularly limited, and sterilization treatment may be performed after freeze-drying.
  • a sterilization method for example, E.O. Methods, such as gas sterilization, gamma-ray sterilization, and E-Beam sterilization, are mentioned.
  • the bone graft material according to the present invention is manufactured by the above-described manufacturing method and is composed of only demineralized bone. Since the bone graft material according to the present invention is composed only of demineralized bone, it has excellent bone induction ability and biocompatibility.
  • the bone graft material according to the present invention is composed of only demineralized bone and manufactured by introducing the above-described demineralized bone-derived carrier, it has high durability, high compressive strength, and excellent shape retention.
  • the bone graft material according to the present invention can be used as a bone filling material, bone bonding material, implant, and bone scaffold.
  • the bone graft material according to the present invention can be suitably used as a support for bone angle adjustment and bone tissue regeneration, as in High Tibial Osteotomy (HTO), foot & ankle surgery, etc. .
  • HTO High Tibial Osteotomy
  • the shape of the bone graft material according to the present invention is not particularly limited, but is preferably a block shape, and more preferably a wedge shape as shown in FIGS. 1 and 2 .
  • the size is not particularly limited. Referring to FIG. 2, for example, the width (W) is 10 mm or more and 50 mm or less, the length (L) is 10 mm or more and 50 mm or less, and the height is 1 (H1 ) may be 5 mm or more and 20 mm or less, and the height 2 (H2) may be 2 mm or more and 10 mm or less.
  • This size can be changed by modifying the mold used in manufacturing the bone graft material.
  • the durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate of the bone graft material composed of demineralized bone manufactured by the manufacturing method according to the present invention were measured by the following methods.
  • Block-type bone graft material 1g (W 0 ) prepared in Examples and Comparative Examples was immersed in 100mL of physiological saline and shaken at 100 rpm in an incubator at 37°C. After 3 hours, the remaining bone graft material blocks were collected, freeze-dried, and weighed to determine W t . Then, the durability of the bone graft material block was evaluated by the following formula.
  • a block-type bone graft material with a size of 0.5 cm ⁇ 0.5 cm was prepared, compressed with a universal physical property measuring device (manufactured by Instron, load cell 1 kN, crosshead speed 2 mm/sec), and the load at the strain 10% point was measured.
  • the concentration of calcium remaining in the bone graft material blocks prepared in Examples and Comparative Examples was measured in ppm units using inductively coupled plasma analysis (ICP-OES), and the units were converted into %.
  • ICP-OES inductively coupled plasma analysis
  • 0.1 g of the block-type bone graft material prepared in Examples and Comparative Examples was dissolved in 1 mL of a 4M guanidine solution. This process was performed in an incubator at 4° C. at 100 rpm for 16 hours. The solution was centrifuged at 4000 rpm for 30 minutes and the supernatant was recovered. The recovered solution was dialyzed against distilled water at 4° C. in a Spectrapor 3 tube (Mr 3500 cutoff) and lyophilized. The recovered freeze-dried powder was quantified at UV 450 nm using BMP-2 ELISA kit.
  • Soft tissue was removed from the bone tissue of the cadaver obtained from domestic and foreign tissue banks. This process was performed aseptically using a trimming set in a clean room. Then, the blood and bone marrow were removed using autoclaved water.
  • the dried bones were pulverized using a bone grinder.
  • the grinding and screening conditions were set so that the particle size of the finally obtained demineralized bone powder was 200 ⁇ m or less, and the other conditions were set so that the particle size of the finally obtained demineralized bone powder was 200 ⁇ m or more and 850 ⁇ m or less.
  • the finally obtained demineralized bone powder having a particle size of 200 ⁇ m or less is described in [1-2. Production of demineralized bone-derived carrier], and the finally obtained demineralized bone powder having a particle size of 200 ⁇ m or more and 850 ⁇ m or less is described in [1-3. Manufacture of block-type bone graft material].
  • the pulverized bone powder was put in 70% ethanol, treated at 15-25°C for 3 hours, put in 3% hydrogen peroxide, treated for 1 hour, and then washed three times for 10 minutes each time using sterile purified water. After washing, the bone powder was put into 70% ethanol again, treated at 15 ⁇ 25 °C for 1 hour, and then dried for 2 hours or more in a sterile workbench.
  • the dried bone powder was treated with a demineralization solution (acidic solution, 0.5N HCl) at 15 to 25° C. for 6 hours.
  • a demineralization solution acidic solution, 0.5N HCl
  • the demineralized bone powder was washed three times with sterilized purified water for 10 minutes each time, and then treated with DPBS (phosphate buffer solution) until the pH reached 7.
  • DPBS phosphate buffer solution
  • the neutralized demineralized bone powder was evenly spread on a square dish, packed in an air permeable pouch, put in a freeze dryer and freeze-dried for 36 hours or more to obtain demineralized bone powder having a moisture content of less than 6%.
  • demineralized bone powder by mixing the demineralized bone powder having a particle size of 200 ⁇ m or less and distilled water to prepare a demineralized bone paste.
  • This demineralized bone dough was treated with an autoclave to obtain a demineralized bone-derived carrier.
  • the autoclave treatment was performed for 60 minutes under conditions of a temperature of 121°C and a pressure of 2 kgf/cm 2 .
  • FIG. 3 (a) A photograph of the demineralized bone-derived carrier obtained in Example 1 is shown in FIG. 3 (a).
  • the demineralized bone powder was sufficiently dissolved in water.
  • the viscosity of this demineralized bone-derived carrier obtained in Example 1 was 714.6 Pa ⁇ s.
  • the mixture for forming a bone graft material was prepared by mixing the demineralized bone powder having a particle size of 200 ⁇ m or more and 850 ⁇ m or less obtained through the step. At this time, the mixing ratio of the demineralized bone-derived carrier and the demineralized bone powder was 73:27 by weight.
  • the mixture for forming the bone graft material was put into a mold and molded into a block shape as shown in FIG. 1, and then freeze-dried in a freeze dryer to obtain a block-type bone graft material.
  • FIG. 3(d) a photograph after molding the mixture for forming a bone graft material of Example 1 into a mold is shown in FIG. 3(d).
  • durability and compressive strength were measured by the method described above. The results are shown in Table 1 below.
  • Demineralized bone powder and demineralized bone were prepared in the same manner as in Example 1, except that the mixing ratio of demineralized bone powder and distilled water was changed so that the ratio of demineralized bone powder and distilled water was 20:80 in [Production of demineralized bone-derived carrier].
  • Derived carriers and block-type bone graft materials were prepared, and durability and compressive strength were measured according to the method described above. The results are shown in Table 1 below. On the other hand, the viscosity of the demineralized bone-derived carrier obtained in Example 2 was 790.8 Pa ⁇ s.
  • Fig. 3(b) shows a photograph of the demineralized bone-derived carrier obtained in Example 4
  • Fig. 3(e) shows a photograph after molding the mixture for forming a bone graft material in a mold.
  • the viscosity of the bone-derived carrier was 986.4 Pa ⁇ s.
  • Fig. 3(c) shows a photograph of the demineralized bone-derived carrier obtained in Example 6, and Fig. 3(f) shows a photograph after molding the mixture for forming a bone graft material in a mold.
  • the viscosity of the bone-derived carrier was 1501.9 Pa ⁇ s.
  • a photograph of the demineralized bone-derived carrier obtained in Comparative Example 1 is shown in Fig. 4 (a), and the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 1 was 186.3 Pa ⁇ s.
  • a photograph after molding the mixture for forming a bone graft material of Comparative Example 1 in a mold is shown in FIG. 4(c), and it can be seen that a lot of moisture remains and molding is not performed.
  • FIG. 4(b) a photograph of the demineralized bone-derived carrier obtained in Comparative Example 3 is shown in FIG. 4(b), and in the demineralized bone-derived carrier obtained in Comparative Example 3, undissolved demineralized bone powder remains, and the viscosity can be measured with a rheometer device. It was impossible.
  • a photograph after molding the mixture for forming a bone graft material of Comparative Example 3 in a mold is shown in FIG.
  • the demineralized bone is dissolved in water. It was found that it was suitably dissolved and that the demineralized bone-derived carrier had a viscosity suitable for use as a carrier for producing a bone graft material.
  • the bone graft materials (Examples 1 to 6) according to the present invention have a low residual calcium concentration, a high residual amount of BMP-2, and a high cell proliferation rate, and thus have excellent bone induction ability and biocompatibility. All were found to be excellent.
  • the demineralized bone powder when used beyond the scope of the present invention in the manufacture of a demineralized bone-derived carrier for the manufacture of a bone graft material, the demineralized bone powder is not sufficiently soluble in water, and the obtained demineralized bone-derived carrier is too stiff.
  • Demineralized bone powder was not mixed uniformly, so when manufacturing a block-type bone graft material, it was not molded according to the shape of the mold, and the resulting bone graft material had poor durability and compressive strength, resulting in poor shape retention (Comparative Examples 3 and 4) .
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 0.1N HCl and the demineralization time was changed to 0.5 hour, and the above [1-2.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 5 was 0.06 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 5, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 6 was 0.09 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 5, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 7 was 0.10 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 0.1 N HCl and the demineralization time was changed to 1 hour, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 8 was 0.08 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 8, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 9 was 0.12 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-shaped bone graft material were prepared in the same manner as in Comparative Example 8, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 10 was 0.22 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 0.1 N HCl and the demineralization time was changed to 3 hours, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 11 was 0.12 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 11, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 12 was 0.33 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 11, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 13 was 0.57 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 0.1N HCl, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 14 was 0.69 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 14, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 15 was 0.97 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 14, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 2 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 16 was 1.23 Pa ⁇ s.
  • the demineralization time in the above (1-1-5. Demineralization) was changed to 0.5 hour, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 17 was 105.8 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 17, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 18 was 113.3 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 17, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 19 was 122.0 Pa ⁇ s.
  • the demineralization time in the above (1-1-5. Demineralization) was changed to 1 hour, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 20 was 180.3 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 20, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 21 was 221.8 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Comparative Example 20, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Comparative Example 22 was 386.3 Pa ⁇ s.
  • the demineralization time in the above (1-1-5. Demineralization) was changed to 3 hours, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 7 was 495.8 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 7, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 8 was 669.2 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 7, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 9 was 844.5 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 10 was 910.6 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 3 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 11 was 967.7 Pa ⁇ s.
  • the demineralization conditions and autoclave treatment conditions during the production of demineralized bone powder are the conditions specified in the present invention (the concentration of the acid solution is 0.3N or more and 0.8 or less,
  • the demineralized bone-derived carrier satisfies the following criteria: demineralization time is 2 hours or more and 7 hours or less, autoclave treatment time is 10 minutes or more and 65 minutes or less)
  • the prepared demineralized bone-derived carrier has a viscosity suitable for use as a carrier for preparing a bone graft material, It can be seen that the block-type bone graft material into which the demineralized bone-derived carrier was introduced had low residual calcium concentration, good residual amount of BMP-2, excellent bone induction ability due to high cell proliferation rate, and excellent shape retention ability due to high durability and compressive strength. (Examples 4, 7 to 11).
  • any of the remaining calcium concentration, BMP-2, and cell proliferation rate of the obtained bone graft material It was found that the bone induction ability of the bone graft material was poor because one or more of them were missing, and the demineralized bone-derived carrier did not have a viscosity suitable for use as a carrier for manufacturing a bone graft material, and the shape retention ability of the bone graft material was also poor because sufficient durability and compressive strength were not obtained ( Comparative Examples 17 to 22).
  • the demineralized bone powder is not sufficiently soluble in water, and the bone graft material It was found that carriers derived from demineralized bone suitable for use as carriers for production could not be obtained (Comparative Examples 23 and 24).
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 1.0 N HCl and the demineralization time was changed to 0.5 hour, and the above [1-2.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 12 was 773.9 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 12, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 13 was 800.2 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 12, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 14 was 927.1 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 1.0 N HCl and the demineralization time was changed to 1 hour, and the above [1-2.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 15 was 1013.4 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 15, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 16 was 1058.5 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 15, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 17 was 1193.2 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 1.0 N HCl and the demineralization time was changed to 3 hours, and the above [1-2.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 18 was 1306.0 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 18, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 19 was 1415.4 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 18, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 20 was 1516.3 Pa ⁇ s.
  • the demineralization solution in the above (1-1-5. Demineralization) was changed to 1.0 N HCl, and the above [1-2. Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 4, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 15 minutes.
  • the obtained block-type bone graft material durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 21 was 1670.7 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 21, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 30 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 22 was 1708.4 Pa ⁇ s.
  • Demineralized bone powder, demineralized bone-derived carrier, and block-type bone graft material were prepared in the same manner as in Example 21, except that the autoclave treatment time in [Production of demineralized bone-derived carrier] was changed to 60 minutes.
  • durability, compressive strength, residual calcium concentration, BMP-2 residual amount, and cell proliferation rate were measured according to the above-described methods. The results are shown in Table 4 below.
  • the viscosity of the demineralized bone-derived carrier obtained in Example 23 was 1832.6 Pa ⁇ s.
  • the demineralization conditions and autoclave treatment conditions during the production of demineralized bone powder are the conditions specified in the present invention (the concentration of the acid solution is greater than 0.8N and less than or equal to 1.5N).
  • the prepared demineralized bone-derived carrier has a viscosity suitable for use as a carrier for manufacturing a bone graft material, It was found that the block-type bone graft material into which the demineralized bone-derived carrier was introduced had low residual calcium concentration, good residual amount of BMP-2, excellent bone induction ability due to high cell proliferation rate, and excellent shape retention ability due to high durability and compressive strength. It was possible (Examples 12 to 23).
  • demineralized bone-derived carrier when an acidic solution having a concentration of more than 0.8 N and 1.5 N or less is used in the production of demineralized bone powder, demineralization proceeds sufficiently even in a relatively short time, and the resulting demineralized bone is produced. It was found that the bone graft material containing the derived carrier had sufficient durability and compressive strength.

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  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
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  • Dermatology (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
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  • Orthopedic Medicine & Surgery (AREA)
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Abstract

La présente invention concerne un procédé de préparation d'un matériau de greffe osseuse qui consiste en un os déminéralisé uniquement et un matériau de greffe osseuse ainsi préparé. Selon la présente invention, par un procédé de préparation comprenant : une étape (a) d'obtention de poudre osseuse déminéralisée par déminéralisation de poudre osseuse ; une étape (b) de préparation d'une pâte osseuse déminéralisée par mélange de l'os déminéralisé et de l'eau à un rapport prédéterminé ; et une étape (c) de traitement de la pâte osseuse déminéralisée dans un autoclave, un support dérivé d'os déminéralisé pour préparer un matériau de greffe osseuse est obtenu, et un procédé de préparation d'un matériau de greffe osseuse, comprenant : une étape (d) de préparation d'un mélange pour former un matériau de greffe osseuse par mélange du support dérivé d'os déminéralisé pour préparer un matériau de greffe osseuse et de la poudre osseuse déminéralisée ; et une étape (e) de lyophilisation du mélange pour former un matériau de greffe osseuse. Selon le procédé de préparation décrit ci-dessus de la présente invention, un support dérivé d'os déminéralisé ayant une viscosité appropriée pour une utilisation en tant que support pour préparer une charge osseuse peut être préparé. De plus, en introduisant le support dérivé d'os déminéralisé, un matériau de greffe osseuse constitué de l'os déminéralisé uniquement et ayant en conséquence une excellente induction osseuse et une excellente biocompatibilité ainsi qu'une excellente rétention de forme est fourni.
PCT/KR2022/009439 2021-07-22 2022-06-30 Matériau de greffe osseuse comprenant un support dérivé d'os déminéralisé introduit et son procédé de préparation WO2023003209A1 (fr)

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KR1020210096608A KR102527814B1 (ko) 2021-07-22 2021-07-22 탈회골 유래 담체가 도입된 골이식재 및 그 제조 방법

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060280803A1 (en) * 2004-10-14 2006-12-14 Mukesh Kumar Method and apparatus for repairing bone
KR20100134306A (ko) * 2009-06-15 2010-12-23 한스바이오메드 주식회사 탈회골(ｄｂｍ)을 포함하는 골 충진재의 제조방법
WO2021050901A1 (fr) * 2019-09-11 2021-03-18 Warsaw Orthopedic, Inc. Fibres de matrice osseuse déminéralisée, procédés de production et d'utilisation associés
US20210196858A1 (en) * 2019-12-31 2021-07-01 Helena Lovick Shapeable demineralized bone matrix products and method of manufacture thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2416481C (fr) * 2000-07-19 2011-02-15 Osteotech, Inc. Implant osseux et son procede de fabrication
KR101817002B1 (ko) 2016-06-10 2018-01-09 조선대학교산학협력단 압축성형 몰드를 이용한 베타-트리칼슘포스페이트계 합성골 웨지의 제조방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060280803A1 (en) * 2004-10-14 2006-12-14 Mukesh Kumar Method and apparatus for repairing bone
KR20100134306A (ko) * 2009-06-15 2010-12-23 한스바이오메드 주식회사 탈회골(ｄｂｍ)을 포함하는 골 충진재의 제조방법
WO2021050901A1 (fr) * 2019-09-11 2021-03-18 Warsaw Orthopedic, Inc. Fibres de matrice osseuse déminéralisée, procédés de production et d'utilisation associés
US20210196858A1 (en) * 2019-12-31 2021-07-01 Helena Lovick Shapeable demineralized bone matrix products and method of manufacture thereof

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