KR20020011164A - Method for Manufacturing bone graft - Google Patents

Method for Manufacturing bone graft Download PDF

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KR20020011164A
KR20020011164A KR1020000044494A KR20000044494A KR20020011164A KR 20020011164 A KR20020011164 A KR 20020011164A KR 1020000044494 A KR1020000044494 A KR 1020000044494A KR 20000044494 A KR20000044494 A KR 20000044494A KR 20020011164 A KR20020011164 A KR 20020011164A
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bone
washed
dried
powder
graft material
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KR1020000044494A
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Korean (ko)
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엄인웅
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이건일
주식회사 휴먼텍
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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
    • 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
    • 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
    • A61F2/2803—Bones for mandibular reconstruction
    • A61F2002/2807—Chin implants
    • 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

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

Abstract

PURPOSE: A method for preparing bone transplanter is provided to obtain an excellent bone transplantation effect without occurring an inflammation and an immune rejection reaction after transplanting. CONSTITUTION: In the method for preparing bone transplanter, allogenic bone is collected, and soft tissue is removed from the bone. The bone is pulverized, washed, and classified into bone pieces and bone powders. The split bone pieces or bone powders are washed and defatted using ethanol and ether, respectively. The bone is dried at room temperature, and demineralized using hydrochloric acid. The demineralized bone is washed with saline solution or phosphate buffered saline, and further washed with ethanol and ether. The washed bone is dried at room temperature and lyophilized. The lyophilized bone is further classified by sizes to obtain bone transplanter.

Description

골이식 물질의 제조방법{Method for Manufacturing bone graft}Method for Manufacturing bone graft

본 발명은 신규한 골이식 물질에 관한 것으로, 보다 상세하게는 동종골을 채취 및 처리함으로써 우수한 골이식효과를 얻을 수 있는 신규한 골이식 물질 및 그 제조방법에 관한 것이다.The present invention relates to a novel bone graft material, and more particularly to a novel bone graft material and a method for producing the same bone graft effect can be obtained by collecting and treating allogeneic bone.

상악골이나 하악골에는 양성종양과 악성종앙이 빈발하고 있으며 이에 대한 외과적 치료법으로는 아직까지 병소부가 포함된 골을 절제하는 방법이 많이 시행되고 있다. 외과적인 결손부위가 커질수록 저작, 교합, 발음 등에 지장을 초래하게 되어 심미적으로도 불량하고 일상생활에 많은 불편을 주게된다. 따라서 결손부분은 정상적인 기능과 심미적인 관점에서 재건해주어야 하는데 이와 관련해서는 동종골, 이종골 및 합성이물질 등의 골대체물질의 개발이 활발하게 진행되고 있다.Benign and malignant tumors are frequently found in the maxilla and mandible. Surgical treatments for this are frequently performed to remove the bone including the lesion. The larger the surgical defect, the greater the disruption in chewing, occlusion, pronunciation, etc., resulting in poor aesthetics and inconvenient daily life. Therefore, the defect should be reconstructed from the normal function and aesthetic point of view. In this regard, development of bone substitutes such as allogeneic bone, xenograft, and synthetic foreign body is actively progressing.

신서 자가골이식(fresh autograft)는 일반적으로 면역학적으로 안정적이며 혈관의 신생이 빠르고 숙주와의 친화력이 우수하기 때문에 정상 골 조직을 형성하려는 재생의 관점에서 다른 골 이식보다 우수하며 현재 가장 좋은 방법으로 이용되고 있다. 그러나 자가골 이식은 부가적인 수술을 해야하고 이식에 필요한 모양, 크기, 골의 양과 관련된 수술상의 제약 때문에 임상적인 응용이 제한되고 있다.Fresh autografts are generally immunologically stable, their angiogenesis is fast and their affinity with the host is superior to other bone grafts in terms of regeneration to form normal bone tissue and are currently used as the best method. It is becoming. Autologous bone grafts, however, require additional surgery and are limited in clinical applications due to surgical constraints related to the shape, size, and amount of bone needed for transplantation.

이러한 신선 자가골 이식의 단점을 보완하기 위한 것으로 병소가 있는 부위의 골을 절제한 후 냉동 처리하여 재이식하는 방법이 사용되기도 하지만 종양의 재발이나 괴사가 종종 발생하기 때문에 처리방법에 대한 많은 연구가 진행되고 있다. 단순 냉동처리골에 대한 연구들을 살펴보면 마르시아니(Marciani) 박사는 개에서 부분 절제술에 의하여 제고된 하악골을 액화질소로 냉동처리한 후 다시 제자리에 넣음으로써, 처리하지 않은 본래의 하악골과 결합시키는 것이 가능하다는 것을 증명하였다. 이러한 기술을 그 자리에서 냉동시키는 것보다 외부에서 냉동시키므로써 인접 연조직에 대한 부적절한 온도상승으로 인한 창상 열개의 위험성을 감소시키면서 종양골내의 종양세포를 파괴시키는 효과가 있다고 보고하였다(Marciani RD, Bowden CM, Reimplantation of freeze-treated mandibular bone, J. Oral. Surg., 33: 261, 1975).In order to make up for the drawbacks of the fresh autologous bone graft, a method of resection and freezing after resection of the bone at the site of the lesion is used. However, many studies on the treatment method are ongoing because tumor recurrence or necrosis often occurs. It is becoming. In studies of simple frozen bones, Dr. Marcani was able to bind the untreated original mandible by freezing it with liquid nitrogen and then putting it back in place by liquid nitrogen in dogs. Proved. It has been reported that freezing this technique externally, rather than freezing in situ, has the effect of destroying tumor cells in the tumor bone while reducing the risk of wound degeneration due to inappropriate temperature rise in adjacent soft tissues (Marciani RD, Bowden CM). , Reimplantation of freeze-treated mandibular bone, J. Oral. Surg., 33: 261, 1975).

한편, 동종골 이식(Homologous grafts, Allografts, Allogenic grafts)은 같은 종의 다른 개인에게 골이식을 하는 것으로서, 동종골은 면역학적 부작용으로 인한 면역거부반응이 나타나 그 동안 많은 연구들이 주로 동종골의 면역학적 억제와 골형성 활성을 갖도록 처리하는 방법을 찾는데 주력했다.(Langer F, Czitrom A, Pritzker KP, Gross AE; The immunogenecity of fresh an frozen allogenic bone. J Bone Joint Surg 1975; 57A: 216-220). 이식재의 항원성을 변경시킴으로써 숙주 면역반응이 자극받지 않도록 하는 방법으로는 끓이기, 단백질 제거, 화학약품사용, 냉동, 동결건조, 방사선 조사 및 건열법 등이 사용되어 왔다.Allografts (Homologous grafts, Allografts, Allogenic grafts) are bone grafts to other individuals of the same species, and allogeneic bones show immunorejection due to immunological side effects. The focus was on finding ways to treat osteogenic activity (Langer F, Czitrom A, Pritzker KP, Gross AE; The immunogenecity of fresh an frozen allogenic bone. J Bone Joint Surg 1975; 57A: 216-220). Boiling, protein removal, chemical use, freezing, lyophilization, irradiation and dry heat methods have been used to prevent the host immune response from being stimulated by altering the antigenicity of the implant.

이식제를 끓이거나 화학적으로 처리하는 것은 세포를 죽이고 항원성을 감소시키는 반면 이 방법은 어떤 골형성 유도기능도 파괴시킬 수 있는데 이는 골내에 존재하는 골형성 단백질(Bone Morphogenic Protein: BMP)을 변질시킴으로써 유발될 수 있다. 끓이기, 건조법 또는 화학처리법에 의한 처리는 이식재 내의 유기요소들을 응고시키기 때문에 숙주가 일련의 세포반응을 통해 그들을 제거하는 것은 매우 어렵다. 골의 방사선 조사는 대개 15 KGy를 사용하고 있고 멸균과 골내 함유된 항원을 파괴시키는 방법으로 사용되어 오고 있으나 멸균에 필요한 조사량이 골의 물리적 성질을 변화시키거나 골형성 유도를 저해할 수 있음을 고려해야한다.(Spengos MN. Irradiated allogenic bone grafts in the treatment of odontogenic cysts. J Oral Surg 1974; 32: 674-678)Boiling or chemically treating the implant kills cells and reduces antigenicity, while this method can destroy any bone induction function by altering Bone Morphogenic Protein (BMP) in the bone. May be induced. Because boiling, drying, or chemical treatment causes coagulation of organic elements in the implant, it is very difficult for the host to remove them through a series of cellular reactions. Irradiation of bone is usually 15 KGy and has been used as a method of sterilization and destruction of antigens contained in bone, but it should be considered that the amount of irradiation required for sterilization may change the physical properties of bone or inhibit the induction of bone formation. (Spengos MN.Irradiated allogenic bone grafts in the treatment of odontogenic cysts. J Oral Surg 1974; 32: 674-678)

현재 주로 사용되는 동종골은 대부분 동결건조법에 의해 처리된다. 동결건조(freeze-drying, lyophilization)와 단순동결(deep-freezing)은 동물실험 모델에서 골이식의 항원성을 감소시키는 것으로 알려져 있고 최근 동결건조한 동종 피질골 토끼실험 모델에서 면역반응이 거의 나타나지 않은 것으로 확인되었다.Most commonly used allografts are currently treated by lyophilization. Freeze-drying (lyophilization) and deep-freezing have been shown to reduce the antigenicity of bone grafts in animal models and have shown little immune response in recent freeze-dried allograft rabbit models. It became.

골이식후 재생에 관한 개념은 이식물이나 매식체의 이행성골대체, 융합, 골유도, 골전도 및 면역거부 등의 개념을 포함한다. 동종골의 이행성 골대체는 골화세포로 분화되고 유골기질을 생성하는 원시성 간엽세포를 공급하는 혈관이 이식체 안에서 성장하는 것에 의존한다. 이식 후에 나타나는 파골세포들과 염증세포들은 숙주에 의해서 자라 들어온 혈관에 의해 운반된다. 골의 실활기질은 혈액이 지나가는 골내 영양관들의 주위로부터 천천히 흡수된다. 실활 골조직이 이식체의 어떤 부위로부터 흡수되는 동안 골형성 간엽세포들에 의해 증여부의 다른 골주표면으로 신생골이 형성된다. 골흡수와 골침착의 유동성은 동종골이 완전히 대치될 때까지 계속되며 이 과정을 이행성 골대체라고 한다. 융합이란 이식물과 숙주사이의 결합을 위해 수여부에서 축적된 새로운 골이 이식 골조직을 둘러싸고 얽히는 과정이다. 이행성 골대체가 일시적이고 공간적인 재생활동인 반면 골융합은 초기 염증반응, 골형성 전구세포증식, 골유도와 골전도 모두를 포함하는 재생의 전과정이며 기전이라 할 수 있다. 융합의 마지막 단계는 이식시 비활성의 골조직이 수여부의 재형성된 층판골로 완전히 둘러싸이는 과정이다. 골전도는 수여부로부터 이식골체 내로확장되는 모세혈관과 혈관주위조직 및 결정성 골형성 전구세포가 성장을 통하여 수여부로부터의 신생골이 형성되면서 확장해 들어간다. 이때 골이식은 수동적인 작용만 하고 골전도는 세라믹이나 플라스틱 같은 매식물질의 흡수가 없는 비생체 재성화되어 연골이나 골을 형성하는 과정이다. 골유도는 유도물질의 영향으로 유도성 골형성전구세포가 활성화되어 연골이나 골을 형성하는 과정이다. 골형성단백질로 알려진 골유도 인자의 효과는 실험실적으로 알려져 있으며 이 단백질은 소수성 당단백질이고 다종간에 골형성반응을 일으킨다고 알려져 있다.Concepts of regeneration after bone graft include concepts such as transplantation of the implant or media, bone fusion, bone induction, bone conduction, and immunorejection. Allogeneic transitional bone substitutes rely on the growth of blood vessels within the graft that supply primitive mesenchymal cells that differentiate into osteoclasts and produce an osteoplasm. The osteoclasts and inflammatory cells that appear after the transplant are carried by blood vessels that have grown in by the host. Bone degeneration of the bone is slowly absorbed from around the bone canal through which blood flows. New bone is formed on the other bone surface of the donor by the osteogenic mesenchymal cells while the inactivated bone tissue is absorbed from any part of the implant. The fluidity of bone resorption and bone deposition continues until allogeneic bone is completely replaced and this process is called transitional bone replacement. Fusion is the process by which new bone accumulates at the recipient site for binding between the implant and the host and entangles the transplanted bone tissue. Transitional bone replacement is a temporary and spatial regenerative activity, while bone fusion is the whole process and mechanism of regeneration including both early inflammatory response, osteogenic progenitor cell proliferation, both bone induction and bone conduction. The final stage of fusion is the process in which the inactive bone tissue at the time of implantation is completely surrounded by the recipient's remodeled lamellar bone. Bone conduction expands as capillaries, perivascular tissue, and crystalline osteogenic progenitor cells grow from the donor site into the graft bone, forming new bone from the donor site. At this time, bone graft is only a passive action, and bone conduction is a process in which cartilage or bone is formed by non-living regeneration without absorption of a material such as ceramic or plastic. Osteoinduction is the process of activating inducible osteogenic progenitor cells to form cartilage or bone under the influence of inducers. The effects of osteoinductive factors known as osteoblastic proteins are known experimentally, and the proteins are hydrophobic glycoproteins and are known to cause osteogenic reactions among various species.

Reddi는 동종골 이식에서 골유도가 여러 단계를 거치는 과정이라고 가정하였는데 초기 화학주성, 피브로넥틴 결합, 간엽세포의 증식 및 혈관증식으로 시작한 후 연골아세포-골아세포 분화와 석회화과정으로 이어진다고 하였다(Reddi AH. Regulation of cartilage and bone differentiation by bone morphogenetic protein. [Review]. Curr Opin Cell Bion 1992; 4: 851-855). 기질유도성 골형성과정 동안 일련의 용해성 기질인자들이 앞서 설명한 각각의 단계를 연결하는데, 그중 가장 중요한 인자가 골형성 단백질이다. 따라서 이러한 골형성 단백질을 안정된 형태로 유지하면서 골기질의 골유도성을 최대로 유지시키는 것이 동종골 이식에 있어서 가장 중요한 관건이라 할 수 있다.Reddi assumes that bone induction is a multi-step process in allogeneic bone grafts, beginning with early chemotaxis, fibronectin binding, mesenchymal cell proliferation and vascular proliferation, followed by chondrocyte-osteoblast differentiation and calcification (Reddi AH). Regulation of cartilage and bone differentiation by bone morphogenetic protein. [Review] .Curr Opin Cell Bion 1992; 4: 851-855). During the matrix-induced bone formation process, a series of soluble matrix factors connect each of the steps described above, the most important of which is the osteogenic protein. Therefore, maintaining the bone formation protein in a stable form while maintaining the bone induction of bone substrate to the maximum can be said to be the most important key in allogeneic bone graft.

본 발명의 목적은 동종골을 이용하여 보다 우수한 이식효과를 나타낼 수 있는 신규한 골이식 물질을 제조하는 방법을 제공하는 것이다.It is an object of the present invention to provide a method for producing a novel bone graft material that can exhibit a better transplantation effect using allogeneic bone.

본 발명의 또 다른 목적은 상기 제조방법으로부터 다양한 용도의 골이식 물질을 제공하는 것이다.Another object of the present invention to provide a bone graft material for various uses from the manufacturing method.

도 1은 상악골내의 골낭을 제거한 후 골결손부위에 본 발명에 따르는 골이식 물질을 넣은 후 찍은 방사선 사진.Figure 1 is a radiograph taken after the bone graft material according to the invention in the bone defect after removing the bone sac in the maxilla.

도 2a 및 도 2b는 상악골 내의 골낭을 제거한 후 골결손 부위에 본 발명에 따르는 골이식 물질을 넣은 직후 및 6개월 후 치유된 사진.Figure 2a and Figure 2b is a picture healed immediately after and 6 months after the bone graft material according to the present invention in the bone defect after removing the bone sac in the maxilla.

도 3a 내지 도 3c는 상악골내의 치근단 골낭, 치근단 골낭을 제거한 후 본 발명에 따르는 골이식 물질을 넣은 직후 및 6개월후 치유 상태의 방사선 사진.Figure 3a to 3c is a radiograph of the healing state immediately after and 6 months after the addition of the bone graft material according to the present invention after removal of the root fracture, the root fracture in the maxilla.

도 4a 및 도 4b는 하악골내에서 치아를 발거한 후 인공치아를 악골에 매식할 때에 인공치아의 골결손 부위에 본 발명에 따르는 골이식 물질을 넣은 직후 및 6개월후의 방사선 사진.Figures 4a and 4b are radiographs immediately after and 6 months after placing the bone graft material according to the present invention to the bone defect site of the artificial tooth when the artificial tooth is buried in the jaw bone after extraction of the tooth in the mandible.

도 5a 및 도 5b는 하악골 내에서 병소부위의 치아를 발거한 후 골결손부위 및 여기에 본 발명에 따르는 골이식 물질을 넣은 후 6개월 뒤에 촬영한 방사선 사진.Figures 5a and 5b is a radiograph taken 6 months after the bone defects and the bone graft material according to the invention after the extraction of the teeth of the lesion in the mandible.

상기 목적을 달성하기 위하여 본 발명에 따르는 골이식 물질의 제조방법은 동종골을 채취하여 연조직을 제거하는 단계; 상기 뼈를 분쇄하여 세척하며 골편과 골분말로 1차 분류하는 단계; 상기 1차 분류된 골편 또는 골분말을 각각 세척하는 단계; 상기 세척된 골편 또는 골분말을 각각 에탄올 및 에테르를 이용하여 탈지하는 단계; 상기 탈지된 뼈를 실온에서 건조하는 단계; 상기 건조된 뼈를 염산을 이용하여 탈회하는 단계; 상기 탈회된 뼈를 세척후 에탄올 및 에테르를 이용하여 세척하는 단계; 상기 세척된 뼈를 실온 건조후 냉동건조시키는 단계; 및 상기 건조된 뼈를 크기별로 2차 분류하는 단계로 이루어지는 것을 특징으로 한다.Method for producing a bone graft material according to the present invention to achieve the above object comprises the steps of removing the soft tissue by collecting allogeneic bone; Crushing and washing the bone and primary sorting into bone fragments and bone powder; Washing the primary sorted bone fragments or bone powder, respectively; Degreasing the washed bone fragments or bone powder using ethanol and ether, respectively; Drying the degreased bone at room temperature; Demineralizing the dried bone with hydrochloric acid; Washing the demineralized bone with ethanol and ether after washing; Lyophilizing the washed bone after room temperature drying; And second classifying the dried bone by size.

본 발명에 따르는 골이식 물질의 원료인 동종골은 사람의 사체로부터 채취된 것을 사용할 수 있다. 예컨대 기증된 시신에서 채취된 골조직을 -70℃ 이하의 냉동고에서 삼중 포장된 상태로 보관된 것을 사용하는 것이 바람직하다.Allogeneic bone, which is a raw material of bone graft material according to the present invention, may be one obtained from a human body. For example, it is preferable to use a bone tissue collected from a donated body stored in a triple package in a freezer below -70 ℃.

본 발명에 따르는 골이식 물질의 재조 방법에 있어서 연조직을 제거하는 단계에서 연조직의 제거가 부적절하게 이루어질 경우 최종 제품에서 연조직이 잔여물질로 존재하게 되므로 특별한 주의를 기울여 최대한으로 연조직을 제거하는 것이 바람직하다.In the manufacturing method of the bone graft material according to the present invention, if the soft tissue is inadequately removed in the step of removing the soft tissue, soft tissue is present as a residual material in the final product, so it is preferable to remove the soft tissue to the maximum with special care. .

본 발명에 따르는 골이식 물질의 제조방법에 있어서 탈지(defatting) 과정은에탄올이나 에테르에 담궈서 각각 1~2시간 동안 교반시킴으로써 수행된다. 이때 에탄올로 세척을 할 경우에는 탈수(dehydration)효과도 동시에 거둘 수 있다.In the method for producing a bone graft material according to the invention the defatting process is carried out by immersing in ethanol or ether and stirring for 1 to 2 hours, respectively. At this time, when washing with ethanol can also achieve a dehydration (dehydration) effect at the same time.

본 발명에 따르는 골이식 물질의 제조방법에서 분쇄과정은 미세골분쇄기 또는 호모지나이져(homogenizer) 등을 이용할 수 있으며, 이때 원료물질이 변질되지 않게 하기 위해서는 분쇄과정에서 온도가 상승되지 않도록 주의한다.In the method for producing a bone graft material according to the present invention, the grinding process may use a fine bone grinder or a homogenizer, and at this time, in order not to deteriorate the raw material, care should be taken not to increase the temperature during the grinding process.

본 발명에 따르는 골이식 물질의 제조방법에서 1차 분류과정은 뼈를 분쇄하면서 채(seive) 위에서 골편(bone chip)과 골분말(bone powder)로 분류하는 과정이다. 이때 골편의 크기는 1mm 이상의 입자크기를 갖도록 하는 것이 바람직하다. 상기와 같이 분류된 골편과 골분말은 생리식염수로 세척을 반복한다.In the method for producing a bone graft material according to the present invention, the primary classification process is a process of classifying bone chips and bone powder on a sieve while grinding bone. At this time, the size of the bone piece is preferably to have a particle size of 1mm or more. The bone fragments and bone powder classified as described above are washed with physiological saline.

분류된 동종골 분말 및 칩은 0.5N HCl에 약 1시간 30분 동안 침지시켜 탈회(demineralization)시킨다. 탈회 횟수는 입자의 크기, 용도에 따라 1회 이상으로 하며, 입자가 작은 분말의 경우는 염산 접촉면적이 상대적으로 많기 때문에 칩에 비하여 탈회 횟수를 적게 할 수 있다.Sorted allogeneic powders and chips are demineralized by soaking in 0.5N HCl for about 1 hour 30 minutes. The number of times of deliming is one or more times depending on the size and use of the particles. In the case of small particles, the number of times of deliming can be reduced compared to chips because the contact area of hydrochloric acid is relatively large.

탈회를 거친 분말 또는 칩은 식염수나 인산염 완충용액(phosphate buffered saline)으로 세척한 다음 에탄올 및 에테르에 각각 1~2 시간씩 침지하여 세척한다. 이는 분말 또는 칩 형상의 동종골을 다시 한번 탈지시킴과 동시에 멸균시키는 효과를 거둘 수 있다.The demineralized powder or chips are washed with saline or phosphate buffered saline and then immersed in ethanol and ether for 1-2 hours each. This can achieve the effect of degreasing at the same time degreasing powder or chip allograft once again.

상기 탈지된 골편 또는 골분말은 실온에서 하룻밤 동안 건조한 다음, 냉동건조시킨다. 이때 냉동건조는 -70℃의 온도에서 약 48시간 동안 수행하는 것이 바람직하다.The degreased bone fragment or bone powder is dried overnight at room temperature and then lyophilized. The freeze-drying is preferably performed for about 48 hours at a temperature of -70 ℃.

본 발명에 따르는 골이식 물질의 제조방법에서 2차 분류 과정은 채를 이용하여 냉동 건조된 골분말을 입자 크기 별로 분류하는 과정이다. 예컨대, 212㎛ 이하, 212~500㎛, 500~700㎛ 및 700~1000㎛ 입자크기의 골분말로 각각 분류할 수 있다.In the method for preparing bone graft material according to the present invention, the secondary classification process is a process of classifying the freeze-dried bone powder by particle size using a shaft. For example, it can be classified into bone powder of 212㎛ or less, 212 ~ 500㎛, 500 ~ 700㎛ and 700 ~ 1000㎛ particle size, respectively.

상기 분류된 골편 또는 골분말은 멸균된 바이알에 분주하여 통상의 방법대로 멸균 포장한다. 예컨대, 에틸렌 옥사이드(ethylene oxide) 가스소독을 실시하여 무균상태에서 포장하고 통상의 멸균 방법으로 소독한다. 또한, 필요에 따라서 에틸렌 옥사이드 가스소독이나 방사선 소독을 추가로 수행할 수 있다.The classified bone fragments or bone powders are dispensed into sterile vials and sterilely packaged according to a conventional method. For example, ethylene oxide gas sterilization is carried out, packaged in a sterile state, and sterilized by a conventional sterilization method. In addition, ethylene oxide gas sterilization or radiation disinfection may be further performed as necessary.

이상과 같은 제조과정을 거쳐 제조된 본 발명에 따르는 골이식 물질은 낭적출환자, 발치 및 인공치아 매식시 골결손부위 및 치주질환에 의한 치조골 결손부위에 사용한 결과 우수한 치유 능력을 보였다.The bone graft material according to the present invention prepared through the manufacturing process as described above showed excellent healing ability when used in alveolar bone defect site and bone defect site at the time of foci extraction, extraction and artificial tooth implantation.

이하 실시예를 통해 본 발명에 따르는 골재생 촉진제에 대하여 보다 상세하게 설명한다.Hereinafter, the bone regeneration accelerator according to the present invention will be described in more detail with reference to the following examples.

실시예 1: 골 이식 분말의 제조Example 1: Preparation of Bone Graft Powder

-70℃ 이하의 냉장고에서 삼중 포장한 상태로 보관되어 있는 기증된 시신으로부터 채취된 골조직을 해동시켰다. 작업에 들어가기 전에 스왑 배양(swab culture)를 시행한 다음, 멸균된 작업대 위에 해동된 뼈를 꺼내어 멸균된 생리식염수로 잘 세척하고 혈액 및 연조직을 잘 제거한 다음 미세골분쇄기를 이용하여 분쇄하였다. 이때 먼저 몇 개의 조각으로 분쇄하여 연조직 및 혈액제거를 계속하면서 뼈를 분쇄하고 채 위에서 세척과 동시에 1mm 이상의 골편과 그 미만의 골분말로 분류하였다. 분류된 골편 또는 골분말 각각에 멸균된 생리식염수를 1.5l를 넣고 교반하면서 세척하였다. 이 세척과정은 30분마다 생리식염수 용액을 교환하여 4회 시행하였다.Bone tissues were thawed from donated bodies stored in triplicate packaging in a refrigerator below −70 ° C. Swap culture was performed before entering the work, and then thawed bone was removed from the sterilized workbench, washed well with sterile saline solution, blood and soft tissue were removed, and then ground using a fine bone grinder. At this time, the first crushed into a few pieces, while continuing to remove the soft tissue and blood, the bone was crushed and washed at the same time and classified into more than 1mm bone fragments and less than the bone powder. 1.5 l of sterile saline was added to each of the classified bone fragments or bone powder, and the mixture was washed with stirring. This washing process was performed four times by replacing the physiological saline solution every 30 minutes.

세척된 골을 99.9% 에탄올 2l을 넣어 90분간 교반하여 세척한 다음, 2l의 에테르를 넣고 90분간 교반하여 세척한 후 하룻밤 동안 상온에서 건조시켰다.The washed bone was added to 2 l of 99.9% ethanol and washed for 90 minutes, and then, 2 l of ether was added thereto, stirred for 90 minutes, and dried overnight at room temperature.

골편에 대해서는 g당 0.5ml의 0.5N HCl을 넣어 90분간 침지시켜 교반후 세척하는 과정을 반복하여 탈회시키고, 골분말에 대해서는 g당 1ml의 0.5N HCl을 넣어 90분간 침지시켜 교반후 세척하여 탈회시켰다. 탈회과정을 거친 골을 각각 1l의 생리식염수로 30분간 3회 세척한 다음 1l의의 PBS 용액으로 30 분간 세척하였다.For bone fragments, 0.5 ml of 0.5N HCl per gram was added and immersed for 90 minutes, followed by stirring. The process of washing was repeated and demineralized. For bone powder, 1 ml of 0.5N HCl per gram was added and immersed for 90 minutes, stirred and washed. Demineralized. The demineralized bone was washed three times for 30 minutes with 1 l of physiological saline and then for 30 minutes with 1 l of PBS solution.

이상과 같이 처리된 골편 및 골분말에 각각 1.5l의 99.9% 에탄올을 넣고 90분간 교반하면서 세척한 다음, 1.5l의 에테르를 넣고 60분간 침지시켜 교반하면서 세척하였다. 이를 각각 배트(vat)에 고르게 펴서 하룻밤 동안 상온 건조시킨 다음, 건조된 골편 및 골분말을 각각 48-96 시간동안 냉동건조시켰다.1.5 l of 99.9% ethanol was added to the treated bone fragments and bone powder, respectively, and the mixture was washed with stirring for 90 minutes. Then, 1.5 l of ether was added thereto, followed by immersion for 60 minutes, followed by washing. It was evenly spread over each bat and dried at room temperature overnight, and the dried bone fragments and bone powder were freeze-dried for 48-96 hours, respectively.

냉동 건조된 골분말은 건조된 각각의 무게를 측정하고 채와 채 쉐이커(seive shaker)를 사용하여 입자 크기별(212㎛ 이하, 212~500㎛, 500~700㎛ 및 700~1000㎛)로 분류하였다.Freeze-dried bone powder was weighed by each dried and classified by particle size (less than 212㎛, 212 ~ 500㎛, 500 ~ 700㎛ and 700 ~ 1000㎛) using a shake shaker (seive shaker) .

멸균된 바이알에 일정 용량의 골편 또는 골분말을 분주하고, 이때의 골편 또는 골분말의 일부를 취하여 배양하며 바이알 등의 용기에 대해서도 스왑 배양을 실시하였다. 에틸렌 옥사이드 가스로 1차 소독을 실시한 다음 멸균 포장하여 필요에 따라 에틸렌 옥사이드 가스 또는 방사선으로 소독을 반복하였다.A predetermined amount of bone fragments or bone powders were dispensed into sterilized vials, a portion of the bone fragments or bone powders were incubated at this time and cultured, and a swap culture was also performed on containers such as vials. The first disinfection was performed with ethylene oxide gas and then sterilized, and the disinfection was repeated with ethylene oxide gas or radiation as necessary.

실시예 2: 본 발명에 따르는 골이식 물질의 낭적출 환자에 대한 치유능Example 2: Healing ability of the bone graft material according to the present invention for the extraction of patients

본 발명에 따르는 골이식 물질의 치유능을 확인하기 위하여 서울대학교 치과대학 구강악안면 외과에 내원한 환자 총 92명(여자 43명, 남자 49명)을 대상으로 하여 낭 적출후 악골 결손부위에 대한 이식시술을 하였다. 골이식 직후, 6개월 및 12개월 후의 방사선학적 소견을 파노라마 방사선 사진 AUTO-III-ECM 방사선 촬영기(75 kVp, 10mA, 12 sec)을 사용하여 촬영하였다.A total of 92 patients (43 women, 49 men) who visited the Oral and Maxillofacial Surgery of Seoul National University Dental University to confirm the healing ability of the bone graft material according to the present invention transplanted to the jaw defect site after the sac extraction The procedure was performed. Immediately after bone graft, radiological findings at 6 and 12 months were taken using a panoramic radiograph AUTO-III-ECM radiograph (75 kVp, 10 mA, 12 sec).

도 1은 상악골내의 골낭을 제거한 후 골견손부위에 본 발명에 따르는 골이식 물질을 넣은 직후 촬영한 방사성 사진이며, 도2a 및 도2b는 같은 환자에서 골이식 직후 및 6개월 후의 치유된 모습을 찍은 사진이다. 도 3a 내지 3c는 상악골 내에 치근단 골낭(3a)을 제거한 후 본 발명에 따르는 골편을 넣은 후(3b), 수술후 6개월후(3c) 수술부위가 새로 생성된 골로 치유되는 상태를 찍은 방사선 사진이다.1 is a radiographic picture taken immediately after the bone graft in the maxilla was removed after the bone graft material according to the present invention, Figures 2a and 2b is taken in the same patient healed immediately after the bone graft and 6 months later It is a photograph. Figure 3a to 3c is a radiograph showing the state of healing of the newly generated bone after the removal of the apical endocyst 3a in the maxilla (3b), the bone fragment according to the present invention (3b), 6 months after the operation (3c).

상기 실험결과 이식후 후유증으로 동통이나 부종이 적은 비율의 환자에서 나타났으나 이는 일반적인 수술 후의 양상과 같았으며 감염으로 이식골을 제거한 경우는 없었고 그 외에 다른 후유증은 나타나지 않았다.As a result of the post-transplant sequelae, patients with low pain or swelling showed the same symptoms as in the general postoperative course, and none of the grafts were removed due to infection.

실시예 3: 본 발명에 따르는 골이식 물질의 발치와 및 치조골 결손부위에 대한 치유능Example 3: Healing ability against extraction and alveolar bone defect of bone graft material according to the present invention

본 발명에 따르는 골이식 물질의 발치와, 인공치아 매식시 골결손부위 및 치주질환에 의한 치조골 결손부위에 대한 효능을 확인하기 위하여 서울, 인천지역의 인치과에 내원한 상기 증례의 환자 232명(여자 112명, 남자 120명)을 대상으로 육아조직을 골결손부위로부터 완전히 제거한 다음 인공치아를 악골에 매식할 때에 인공치아 주위의 골결손 부위에 본 발명에 따르는 골이식 물질을 넣은 직후와 6개월, 12개월 후를 방사선 사진을 촬영하였다.232 patients (female) of the above case who visited a dental department in Seoul, Incheon in order to verify the extraction of bone graft material and the efficacy of alveolar bone defect caused by periodontal disease during artificial tooth implantation 112 men, 120 men) and 6 months immediately after the granulation tissue was completely removed from the bone defect site and the artificial tooth was embedded in the jaw bone when the bone graft material according to the present invention was placed in the bone defect site around the artificial tooth. Twelve months later, radiographs were taken.

도 4a 및 4b는 하악골에서 치아를 발거한 후 인공치아를 악골에 매식할 때 인공치아 주위의 골 결손부위에 본 발명에 따르는 골이식 물질로서 골분말을 넣은 후 촬영한 방사선사진(4a)과 6개월후 골 결손부위가 잘 치유되어 인공치아가 하악골과 잘 결합된 상태를 보여주는 방사선 사진(4b)이다.Figures 4a and 4b is a radiograph (4a) and 6 taken after placing the bone powder as a bone graft material according to the present invention in the bone defects around the artificial teeth when the artificial teeth are buried in the jaw after removing the teeth in the mandible After 4 months, the bone defect was well healed and the radiograph (4b) shows the artificial teeth combined with the mandible.

도 5a 및 도 5b는 하악골 내에서 병소로 인하여 치아를 발거한 후 골결손부위를 촬영한 방사선 사진(5a)과 6개월후 골결손부위에 인공치아를 매식하면서 본 발명에 따르는 골분말을 넣은 후 골 결손부위가 잘 치유되어 인공치아가 하악골과 잘 결합한 상태를 보여주는 방사선 사진(5b)이다.5a and 5b is a radiographic image (5a) of the bone defects after the extraction of the teeth due to the lesion in the mandible and 6 months after the artificial bones are embedded in the bone defects after the bone powder according to the present invention Radiograph (5b) shows that the bone defect is well healed and that the artificial tooth is combined with the mandible.

상기 실험 결과 이식후 관찰기간동안 환자로부터 특기할 만한 염증소견이나 거부반응은 일어나지 않았으며, 이식후 6개월 후에는 방사선 불투과상이 되어 골화가 촉진되었다는 것을 확인할 수 있었다.As a result, no significant inflammation or rejection was observed from the patient during the post-transplantation observation period, and 6 months after the transplantation, it became radiopaque and promoted ossification.

실시예 4: 골밀도의 변화분석Example 4 Analysis of Changes in Bone Mineral Density

상기 실시예 2 및 3에서의 방사선촬영을 기초로 하여 디지털 영상 분석을 하여 본 발명에 따르는 골이식 물질을 이식한 환부의 골밀도 변화를 분석하였다.Digital image analysis was performed on the basis of radiographic imaging in Examples 2 and 3 to analyze the bone density change of the affected area in which the bone graft material according to the present invention was implanted.

골밀도 분석은 영상처리프로그램 NIH 이미지 1.61(미국 NIH의 Wayne Rasband, zippy.nimh.nih.gov 또는 http://rsb.info.nih.gov/nih-image/)에 의해서분석하였다. 골밀도의 변화는 골결손부위의 흑화도의 변화를 감지함으로써 계측하였고 이를 위하여 회색계조도(백색을 0 흑색을 255)를 사용하였다. 흑화도는 선택된 ROI(region of interest) 내의 평균흑화도를 측정하였다. 정상골과의 골밀도변화를 비교하기 위하여 골결손부위가 아닌 인접하고 치아가 포함되지 않은 정상골부위에서 일정크기(50 pixels x 50 pixels)의 ROI를 선택한 후 평균흑화도를 구하였으며 이를 대조값으로 하여 이에 대한 골결손 부위의 평균흑화도의 비율을 구하여 비교하였다(표 1). 통계처리는 SPSS 7.5 프로그램을 사용하여 스튜던트 t-테스트를 시행하였다.Bone mineral density analysis was analyzed by image processing program NIH image 1.61 (Wayne Rasband, zippy.nimh.nih.gov or http://rsb.info.nih.gov/nih-image/) of NIH, USA. Changes in bone density were measured by detecting changes in the degree of blackening of the bone defect area. Gray gradation (white for black and 255 for black) was used. Blackness was measured as the average blackness within the selected region of interest (ROI). In order to compare the bone mineral density with normal bone, the average blackness was calculated after selecting ROI of a certain size (50 pixels x 50 pixels) in the normal bone area without adjacent bone defects and not including teeth. The ratio of average blackness of the bone defect site was calculated and compared (Table 1). Statistical processing was conducted using Student's t-test using the SPSS 7.5 program.

<표 1>TABLE 1

이식전Before transplant 이식직후Immediately after transplantation 6개월후6 months later 12개월후12 months later 낭적출부위Local extraction site 2.74±0.432.74 ± 0.43 2.69±0.252.69 ± 0.25 2.10±0.31* 2.10 ± 0.31 * 1.65±0.57* 1.65 ± 0.57 * 발치와Extraction and 2.67±0.522.67 ± 0.52 2.61±0.372.61 ± 0.37 2.03±0.24* 2.03 ± 0.24 * 1.55±0.36* 1.55 ± 0.36 * 인공치아매식시Artificial tooth feeding 골결손부위Bone defect 2.53±0.232.53 ± 0.23 2.47±0.362.47 ± 0.36 2.11±0.43* 2.11 ± 0.43 * 1.57±0.61* 1.57 ± 0.61 * 치주질환에 의한 치조골 결손부위Alveolar bone defect due to periodontal disease 2.41±0.392.41 ± 0.39 2.35±0.212.35 ± 0.21 2.03±0.16* 2.03 ± 0.16 * 1.69±0.31* 1.69 ± 0.31 *

상기 표 1로부터 확인되는 바와 같이 본 발명에 따르는 골이식 물질을 시술하기 전후의 골밀도 변화를 정상골부위와 비교한 결과 이식후 6개월 및 12개월에서 이식 직후에 비하여 유의적인 차이를 보였으며, 이는 본 발명에 따르는 골이식 물질에 의해 현저하게 골화가 촉진되었다는 것을 의미한다.As can be seen from Table 1, as a result of comparing the bone density change before and after the bone graft material according to the present invention with the normal bone area, there was a significant difference compared to immediately after transplantation at 6 months and 12 months after transplantation. It is meant that ossification was markedly promoted by the bone graft material according to the invention.

이상에서 살펴본 바와 같이, 본 발명에 따르는 골이식 물질의 제조방법에 의해 제조된 골이식 물질은 이식후 환자들로부터 염증소견이나 면역거부반응을 일으키지 않고 정상 골밀도에 근접하는 정도로 골화를 촉진시키는 것으로 확인되었으며, 따라서 본 발명에 따르는 골이식 물질은 각종 골결손부위의 손상을 치유하기 위하여 사용될 수 있는 매우 유용한 발명이다.As described above, the bone graft material prepared by the method for producing a bone graft material according to the present invention is confirmed to promote ossification to close to normal bone density without causing inflammatory findings or immune rejection reactions from patients after transplantation Therefore, the bone graft material according to the present invention is a very useful invention that can be used to cure damage of various bone defects.

Claims (3)

동종골을 채취하여 연조직을 제거하는 단계; 상기 뼈를 분쇄하여 세척하며 골편과 골분말로 1차 분류하는 단계; 상기 1차 분류된 골편 또는 골분말을 각각 세척하는 단계; 상기 세척된 골편 또는 골분말을 각각 에탄올 및 에테르를 이용하여 탈지하는 단계; 상기 탈지된 뼈를 실온에서 건조하는 단계; 상기 건조된 뼈를 염산을 이용하여 탈회하는 단계; 상기 탈회된 뼈를 세척후 에탄올 및 에테르를 이용하여 세척하는 단계; 상기 세척된 뼈를 실온 건조후 냉동건조시키는 단계; 및 상기 건조된 뼈를 크기별로 2차 분류하는 단계로 이루어지는 것을 특징으로 하여 골이식 물질을 제조하는 방법.Removing allograft to remove soft tissue; Crushing and washing the bone and primary sorting into bone fragments and bone powder; Washing the primary sorted bone fragments or bone powder, respectively; Degreasing the washed bone fragments or bone powder using ethanol and ether, respectively; Drying the degreased bone at room temperature; Demineralizing the dried bone with hydrochloric acid; Washing the demineralized bone with ethanol and ether after washing; Lyophilizing the washed bone after room temperature drying; And classifying the dried bone by size in a second manner. 제 1 항에 있어서,The method of claim 1, 상기 탈회단계는 0.5N 염산에서 약 1시간 30분 동안 침지시키는 것을 특징으로 하는 방법.The deliming step is characterized in that immersed in 0.5N hydrochloric acid for about 1 hour 30 minutes. 제 1 항에 있어서,The method of claim 1, 상기 냉동 건조 단계를 거친 다음 추가로 에틸렌 옥사이드 가스소독을 하는 단계로 이루어지는 것을 특징으로 하는 방법.After the freeze-drying step, and further comprising the step of further sterilizing ethylene oxide gas.
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WO2011016684A3 (en) * 2009-08-06 2011-06-03 Um In Woong Alveolar bone graft processing method and alveolar bone graft therefrom and method for treatment using alveolar bone graft
KR101139337B1 (en) * 2010-02-09 2012-05-03 조선대학교기술지주 주식회사 Grafting material for teeth bone and method of it
WO2012018241A3 (en) * 2010-08-05 2012-05-18 Um In Woong Method for processing bone graft material using teeth, and bone graft material processed thereby
WO2012057454A3 (en) * 2010-10-27 2012-07-26 Park Chang Soo Method for producing a bone transplant material, and bone transplant material produced by same
KR101229435B1 (en) * 2010-08-16 2013-02-05 한스바이오메드 주식회사 Manufacturing Method of Virus Inactivated Bone Powder for Grafting
WO2013108963A1 (en) * 2012-01-17 2013-07-25 Cho Seong-Yong Dental block bone graft and method for manufacturing same
KR101382067B1 (en) * 2013-02-14 2014-04-04 티비엠 주식회사 Bone powder made of own teeth of human being having a crown surpport and manufacturing method thereof
KR101386322B1 (en) * 2013-02-14 2014-04-17 티비엠 주식회사 Bone powder made of own teeth of human being and manufacturing method thereof

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KR100978562B1 (en) * 2008-12-31 2010-08-27 주식회사 코리아본뱅크 Cancellous bone graft substitute and its process
US8409623B2 (en) 2008-12-31 2013-04-02 Korea Bone Bank, Inc. Cancellous bone graft substitute and method of manufacturing the same
CN102470034A (en) * 2009-08-06 2012-05-23 严仁雄 Alveolar bone graft processing method and alveolar bone graft therefrom and method for treatment using alveolar bone graft
EP2462899A4 (en) * 2009-08-06 2013-08-07 In Woong Um Alveolar bone graft processing method and alveolar bone graft therefrom and method for treatment using alveolar bone graft
WO2011016684A3 (en) * 2009-08-06 2011-06-03 Um In Woong Alveolar bone graft processing method and alveolar bone graft therefrom and method for treatment using alveolar bone graft
JP2013500823A (en) * 2009-08-06 2013-01-10 イン ウォン ウン Method of processing alveolar bone graft material and alveolar bone graft material, and treatment method using the above alveolar bone graft material
KR101139337B1 (en) * 2010-02-09 2012-05-03 조선대학교기술지주 주식회사 Grafting material for teeth bone and method of it
CN103068415A (en) * 2010-08-05 2013-04-24 严仁雄 Method for processing bone graft material using teeth, and bone graft material processed thereby
WO2012018241A3 (en) * 2010-08-05 2012-05-18 Um In Woong Method for processing bone graft material using teeth, and bone graft material processed thereby
KR101229435B1 (en) * 2010-08-16 2013-02-05 한스바이오메드 주식회사 Manufacturing Method of Virus Inactivated Bone Powder for Grafting
WO2012057454A3 (en) * 2010-10-27 2012-07-26 Park Chang Soo Method for producing a bone transplant material, and bone transplant material produced by same
CN103200972A (en) * 2010-10-27 2013-07-10 克世摸生物医学有限公司 Method for producing a bone transplant material, and bone transplant material produced by same
US9610383B2 (en) 2010-10-27 2017-04-04 Cosmobiomedicare Co., Ltd. Method for producing a bone transplant material, and bone transplant material produced by same
WO2013108963A1 (en) * 2012-01-17 2013-07-25 Cho Seong-Yong Dental block bone graft and method for manufacturing same
KR101382067B1 (en) * 2013-02-14 2014-04-04 티비엠 주식회사 Bone powder made of own teeth of human being having a crown surpport and manufacturing method thereof
KR101386322B1 (en) * 2013-02-14 2014-04-17 티비엠 주식회사 Bone powder made of own teeth of human being and manufacturing method thereof

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