JP2005110710A - Bone replacing material and its manufacturing method - Google Patents
Bone replacing material and its manufacturing method Download PDFInfo
- Publication number
- JP2005110710A JP2005110710A JP2003344710A JP2003344710A JP2005110710A JP 2005110710 A JP2005110710 A JP 2005110710A JP 2003344710 A JP2003344710 A JP 2003344710A JP 2003344710 A JP2003344710 A JP 2003344710A JP 2005110710 A JP2005110710 A JP 2005110710A
- Authority
- JP
- Japan
- Prior art keywords
- bone
- bone marrow
- tcp
- calcium phosphate
- patient
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 88
- 239000000463 material Substances 0.000 title claims abstract description 60
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims abstract description 79
- 210000001185 bone marrow Anatomy 0.000 claims abstract description 55
- 210000004623 platelet-rich plasma Anatomy 0.000 claims abstract description 52
- 238000000034 method Methods 0.000 claims abstract description 32
- 230000008569 process Effects 0.000 claims abstract description 26
- 229910000389 calcium phosphate Inorganic materials 0.000 claims abstract description 25
- 239000001506 calcium phosphate Substances 0.000 claims abstract description 25
- 235000011010 calcium phosphates Nutrition 0.000 claims abstract description 25
- 210000004369 blood Anatomy 0.000 claims abstract description 12
- 239000008280 blood Substances 0.000 claims abstract description 12
- 238000012258 culturing Methods 0.000 claims abstract description 9
- 238000000605 extraction Methods 0.000 claims description 10
- 238000005470 impregnation Methods 0.000 claims description 6
- 230000000638 stimulation Effects 0.000 claims description 5
- 230000021164 cell adhesion Effects 0.000 abstract description 11
- 230000011164 ossification Effects 0.000 abstract description 11
- 238000005070 sampling Methods 0.000 abstract 3
- 230000007547 defect Effects 0.000 description 16
- 210000000963 osteoblast Anatomy 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 7
- 239000011148 porous material Substances 0.000 description 6
- 238000001356 surgical procedure Methods 0.000 description 6
- 239000002131 composite material Substances 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 210000002798 bone marrow cell Anatomy 0.000 description 4
- DHCLVCXQIBBOPH-UHFFFAOYSA-N Glycerol 2-phosphate Chemical compound OCC(CO)OP(O)(O)=O DHCLVCXQIBBOPH-UHFFFAOYSA-N 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002188 osteogenic effect Effects 0.000 description 3
- 210000003455 parietal bone Anatomy 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 230000009469 supplementation Effects 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 208000006735 Periostitis Diseases 0.000 description 2
- 206010036790 Productive cough Diseases 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000001000 micrograph Methods 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 210000003460 periosteum Anatomy 0.000 description 2
- 210000003802 sputum Anatomy 0.000 description 2
- 208000024794 sputum Diseases 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- -1 β-TCP Chemical compound 0.000 description 2
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 description 1
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 description 1
- 241001314440 Triphora trianthophoros Species 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 230000001936 parietal effect Effects 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Images
Landscapes
- Materials For Medical Uses (AREA)
- Prostheses (AREA)
Abstract
Description
本発明は、人体内の骨欠損部に補填される骨補填材、及びその製造方法に関するものである。 The present invention relates to a bone filling material to be filled in a bone defect portion in a human body and a method for manufacturing the same.
近年、骨腫瘍摘出や外傷等により生じた骨の欠損部に、骨補填材を補填することにより、骨を再生させて骨欠損部を修復することが可能になってきている。骨補填材としては、ハイドロキシアパタイトやリン酸カルシウムが知られているが、体内に異物を残さないとする考え方から、例えばβ−リン酸三カルシウム（β−ＴＣＰ）等、リン酸カルシウムの多孔体からなる足場材が使用されることが多くなってきている。このβ−ＴＣＰを骨欠損部の骨組織に接触させておくと、破骨細胞がβ−ＴＣＰを吸収し、骨芽細胞が新しい骨を形成する、いわゆるリモデリングが行われる。すなわち、骨欠損部に補填された骨補填材は、経時的に自家骨に置換されていくこととなる。 In recent years, it has become possible to regenerate a bone and repair the bone defect by replenishing the bone defect caused by bone tumor extraction, trauma, or the like with a bone filling material. Hydroxyapatite and calcium phosphate are known as bone prosthetic materials, but from the idea that no foreign matter remains in the body, for example, a scaffolding material comprising a porous body of calcium phosphate such as β-tricalcium phosphate (β-TCP) Are increasingly being used. When this β-TCP is kept in contact with the bone tissue of the bone defect portion, so-called remodeling is performed in which osteoclasts absorb β-TCP and osteoblasts form new bone. That is, the bone prosthetic material compensated in the bone defect portion is replaced with the autologous bone with time.
β−ＴＣＰをはじめとするリン酸カルシウムの多孔体としては、例えば特許文献１に記載されている方法によって製造されたものが知られている。このように、所定の気孔率を有する多孔質焼結体とすることで、人体内に埋め込まれ補填された際に、多数の気孔内に十分に骨芽細胞を侵入させて新生骨を形成することができる。
しかし、細胞とβ−ＴＣＰとの接着性に関しては、未だ十分と言えるものではなかった。すなわち、β−ＴＣＰを多孔体として内部に細胞を侵入させ易くしても、気孔内における細胞の細胞接着に時間がかかってしまい、そのため自家骨に置換されるまでに長期間を要しているのが実情であった。こうしたことから、細胞接着をより早期に実現させて、自家骨への置換をより短時間で行い得るようにすることが求められていた。 However, the adhesion between the cells and β-TCP has not been sufficient. That is, even if β-TCP is used as a porous body and cells can easily enter inside, it takes time for the cells to adhere to the cells in the pores, and thus it takes a long time to be replaced with autologous bone. It was the actual situation. For these reasons, it has been demanded that cell adhesion can be realized at an earlier stage and replacement with autologous bone can be performed in a shorter time.
本発明は上記事情に鑑みてなされたもので、補填後の細胞接着を早期に実現させて骨形成を促進し、自家骨への置換を短期間で行い得る骨補填材、及びその製造方法を提供することを目的とする。 The present invention has been made in view of the above circumstances, a bone prosthetic material that can realize cell adhesion after supplementation at an early stage to promote bone formation, and can be replaced with autologous bone in a short period of time, and a method for producing the same. The purpose is to provide.
請求項１に記載の発明は、リン酸カルシウムを足場材とする骨補填材であって、患者の血液から抽出した多血小板血漿が、前記リン酸カルシウムに含有されていることを特徴とする。 The invention according to claim 1 is a bone prosthetic material using calcium phosphate as a scaffold, wherein platelet-rich plasma extracted from the blood of a patient is contained in the calcium phosphate.
請求項２に記載の発明は、請求項１に記載の骨補填材であって、前記患者から採取した骨髄が、前記リン酸カルシウムに含有されていることを特徴とする。 The invention according to claim 2 is the bone grafting material according to claim 1, wherein bone marrow collected from the patient is contained in the calcium phosphate.
請求項３に記載の発明は、リン酸カルシウムを足場材とする骨補填材を製造する方法であって、患者の血液を採取し多血小板血漿を抽出する抽出過程と、該多血小板血漿を前記リン酸カルシウムに混合する混合過程と、を有することを特徴とする。 The invention according to claim 3 is a method for producing a bone grafting material using calcium phosphate as a scaffold, wherein an extraction process of collecting blood from a patient and extracting platelet-rich plasma, and converting the platelet-rich plasma into the calcium phosphate And a mixing process of mixing.
請求項４に記載の発明は、請求項３に記載の骨補填材の製造方法であって、前記抽出過程の前に、前記患者から骨髄を採取する骨髄採取過程と、該骨髄を前記リン酸カルシウムに含浸させる含浸過程と、前記リン酸カルシウムに含浸された前記骨髄を所定時間培養する培養過程と、を有することを特徴とする。 Invention of Claim 4 is a manufacturing method of the bone grafting material of Claim 3, Comprising: The bone marrow collection process which extract | collects a bone marrow from the said patient before the said extraction process, This bone marrow is made into the said calcium phosphate. An impregnation process for impregnation, and a culture process for culturing the bone marrow impregnated with the calcium phosphate for a predetermined time.
請求項５に記載の発明は、請求項４に記載の骨補填材の製造方法であって、前記培養過程において、前記骨髄に超音波刺激を与えることを特徴とする。 The invention according to claim 5 is the method for producing the bone grafting material according to claim 4, wherein ultrasonic stimulation is applied to the bone marrow in the culturing process.
このように、患者の血液から抽出した多血小板血漿をリン酸カルシウムに混合し含有させるようにしているので、多血小板血漿に含有されている天然の形成因子によって、骨形成に関与する細胞（骨芽細胞等）の活性化が促進される。そのため、補填後の骨補填材において細胞接着が早期に実現されて骨形成が促進されるので、自家骨への置換を短期間で行うことができる。 In this way, platelet-rich plasma extracted from the patient's blood is mixed with calcium phosphate so that it is contained, so that cells involved in bone formation (osteoblasts) by natural formation factors contained in platelet-rich plasma. Etc.) is activated. Therefore, since cell adhesion is realized at an early stage and bone formation is promoted in the bone filling material after filling, replacement with autologous bone can be performed in a short period of time.
また、患者から採取した骨髄もリン酸カルシウムに含浸させるようにすれば、骨髄に含有されている骨髄細胞がリン酸カルシウムに播種されるので、予め骨髄細胞を播種した状態の骨補填材とすることができる。そして、この骨髄細胞のリン酸カルシウム上での挙動が、多血小板血漿の形成因子の作用によって、ｉｎ−ｖｉｔｒｏの状態で活性化される。このように、骨髄細胞が活性化された状態となっている骨補填材を補填することができるので、細胞接着が更に早期に、更に的確に実現され、自家骨への置換をより短期間で行うことができる。 In addition, if the bone marrow collected from the patient is also impregnated with calcium phosphate, the bone marrow cells contained in the bone marrow are seeded on the calcium phosphate, so that it can be used as a bone filling material in a state where bone marrow cells are seeded in advance. The behavior of bone marrow cells on calcium phosphate is activated in an in-vitro state by the action of platelet-rich plasma forming factors. In this way, the bone grafting material in which the bone marrow cells are activated can be supplemented, so that cell adhesion can be realized more quickly and more accurately, and replacement with autologous bone can be performed in a shorter period of time. It can be carried out.
更に、患者から多血小板血漿を抽出する、あるいは骨髄を採取するようにしているので、一連の作業を全て骨補填手術の最中において行うことができ、手術中の短時間内に骨補填材の製造を完了し、その後速やかに患者の体内に埋め込み補填することができる。そして、多血小板血漿あるいは骨髄とも、患者から採取したものをまた同一患者の体内に戻すために、拒絶反応等を起こすおそれが無く、極めて信頼性の高い手術を的確に行うことができる。 Furthermore, since platelet-rich plasma is extracted from the patient or the bone marrow is collected, a series of operations can be performed during the bone repair operation, and within a short time during the operation, The manufacture can be completed and then immediately implanted into the patient's body. Since both platelet-rich plasma and bone marrow are collected from the patient and returned to the body of the same patient, there is no risk of rejection and the like, and an extremely reliable operation can be performed accurately.
以下、本発明の実施の形態について説明する。
本実施形態に係る骨補填材の製造方法は、患者の骨欠損部に骨補填材を補填するための手術（骨補填手術）の最中において、骨補填材を製造するものである。
Embodiments of the present invention will be described below.
The method for manufacturing a bone grafting material according to the present embodiment is for producing a bone grafting material during a surgery (bone grafting surgery) for filling a bone defecting part of a patient with a bone grafting material.
先ず手術前の準備作業として、β−リン酸三カルシウム（β−ＴＣＰ）からなる多孔体を、所望の形状、つまり補填される骨欠損部の形状に合致する形状として、予めブロック状に成形しておく。このβ−ＴＣＰは、骨補填材のベースとなる足場材として用いられるものであり、例えば特開平５−２３７１７８号公報に開示されている方法によって製造される。 First, as a preparatory work before surgery, a porous body made of β-tricalcium phosphate (β-TCP) is formed into a block shape in advance so as to have a desired shape, that is, a shape that matches the shape of the bone defect to be compensated. Keep it. This β-TCP is used as a scaffolding material that serves as a base for the bone grafting material, and is manufactured by, for example, a method disclosed in JP-A-5-237178.
次に、手術中の作業として、手術中の患者から血液を採取し、この血液から多血小板血漿（ＰＲＰ）を抽出する（抽出過程）。このＰＲＰの抽出は、例えば公知の遠心分離法により行われる。そして抽出したＰＲＰを、予め製造しておいたβ−ＴＣＰに混合する（混合過程）。このとき、β−ＴＣＰの気孔内にもＰＲＰが十分に行き渡るようにする。 Next, as an operation during surgery, blood is collected from the patient during surgery, and platelet-rich plasma (PRP) is extracted from this blood (extraction process). This extraction of PRP is performed, for example, by a known centrifugation method. Then, the extracted PRP is mixed with β-TCP prepared in advance (mixing process). At this time, PRP should be sufficiently distributed in the pores of β-TCP.
ＰＲＰには、例えばＴＧＦ−β１、ＰＤＧＦ、ＩＧＦ−１といった、種々な天然の形成因子（自家成長因子）が含まれており、これら形成因子によって骨芽細胞等の活性化が促進されることが、わかってきている。このため、こうしたＰＲＰを含有するβ−ＴＣＰを骨補填材として骨欠損部に補填すれば、骨補填材周辺の骨芽細胞等は直ちに活性化され、β−ＴＣＰの気孔内に速やかに侵入していき、β−ＴＣＰへの細胞接着が早期に実現されて、骨形成が促進される。 PRP contains various natural forming factors (autologous growth factors) such as TGF-β1, PDGF, and IGF-1, and activation of osteoblasts and the like is promoted by these forming factors. I know. Therefore, if β-TCP containing such PRP is used as a bone grafting material to fill a bone defect part, osteoblasts and the like around the bone grafting material are immediately activated and quickly penetrate into the pores of β-TCP. As a result, cell adhesion to β-TCP is realized at an early stage, and bone formation is promoted.
なお、このようにＰＲＰを含有した状態のβ−ＴＣＰを、骨補填材として骨欠損部に補填しても良いが、骨髄を混合するようにすれば、より好ましい。こうしたＰＲＰ及び骨髄を含有する骨補填材の製造方法は、上記抽出過程の前に、後述する骨髄採取過程、含浸過程、及び培養過程を有するものである。 It should be noted that β-TCP containing PRP as described above may be supplemented to the bone defect part as a bone grafting material, but it is more preferable to mix bone marrow. Such a method for producing a bone grafting material containing PRP and bone marrow has a bone marrow collection process, an impregnation process, and a culture process, which will be described later, before the extraction process.
先ず、手術中の患者の骨欠損部等から、骨髄を採取し（骨髄採取過程）、この骨髄をβ−ＴＣＰに含浸させる（含浸過程）。このとき、β−ＴＣＰの気孔内にも骨髄が十分に行き渡るようにする。 First, bone marrow is collected from a bone defect or the like of a patient during surgery (bone marrow collection process), and this bone marrow is impregnated with β-TCP (impregnation process). At this time, the bone marrow should be sufficiently distributed in the pores of β-TCP.
そして、骨髄を含浸したβ−ＴＣＰを、骨形成培地に所定時間浸漬することで、骨髄を所定時間培養する（培養過程）。ここで用いる骨形成培地は、デキサメタゾン（Ｄｅｘａｍｅｔｈａｓｏｎｅ）、β−グリセロフォスフェイト（β−ＧＰ）、及びビタミンＣを含有するものである。なお、培養時間すなわち骨形成培地への浸漬時間は、約２時間とすれば、ほぼ十分である。また、ここでいう「培養」とは、細胞の増殖や分化を期待して行うものではなく、骨髄中に含まれている骨芽細胞をβ−ＴＣＰの表面に付着させることを、主たる目的として行うものである。こうすることで、骨芽細胞がβ−ＴＣＰに播種されるので、予め骨芽細胞を播種した状態の骨補填材とすることができる。 Then, by immersing β-TCP impregnated with bone marrow in an osteogenic medium for a predetermined time, the bone marrow is cultured for a predetermined time (culture process). The osteogenic medium used here contains dexamethasone, β-glycerophosphate (β-GP), and vitamin C. It should be noted that the culture time, that is, the immersion time in the bone formation medium, is approximately 2 hours if it is approximately 2 hours. In addition, “culture” as used herein is not performed in the hope of cell proliferation or differentiation, but mainly for attaching osteoblasts contained in the bone marrow to the surface of β-TCP. Is what you do. By carrying out like this, since an osteoblast is seed | inoculated by (beta) -TCP, it can be set as the bone filling material of the state seed | inoculated the osteoblast previously.
なお、この培養過程においては、超音波発生手段を用いて、骨髄に超音波刺激を与えることが好ましい。このような刺激を与えることで、骨形成培地において、β−ＴＣＰと骨髄との初期親和性、細胞の分化・増殖を高めることができる。 In this culturing process, it is preferable to apply ultrasonic stimulation to the bone marrow using ultrasonic wave generating means. By giving such stimulation, initial affinity between β-TCP and bone marrow and differentiation / proliferation of cells can be increased in the osteogenic medium.
このように、ＰＲＰを混合する前に、骨髄をβ−ＴＣＰに含浸させて培養することで、骨髄に含まれている骨芽細胞のβ−ＴＣＰ上での挙動が、ＰＲＰの形成因子の作用によって、ｉｎ−ｖｉｔｒｏの状態で活性化される。このような、ＰＲＰ及び骨髄を含有するβ−ＴＣＰを骨補填材として骨欠損部に補填すれば、上述したように、ＰＲＰの形成因子の作用によって、骨補填材周辺の骨芽細胞は直ちに活性化され、β−ＴＣＰの気孔内に速やかに侵入していき、β−ＴＣＰへの細胞接着が早期に実現される。なおこのとき、β−ＴＣＰ上の骨芽細胞は予め活性化された状態となっているので、これら細胞同士がβ−ＴＣＰ内で速やかに一体化しようとする。その結果、β−ＴＣＰへの細胞接着が、更に早期に且つ的確に実現される。 Thus, before mixing PRP, the bone marrow is impregnated with β-TCP and cultured, so that the behavior of osteoblasts contained in the bone marrow on β-TCP is affected by the action of the formation factor of PRP. Is activated in an in-vitro state. When β-TCP containing PRP and bone marrow is used as a bone grafting material to fill a bone defect, as described above, osteoblasts around the bone grafting material are immediately activated by the action of the PRP forming factor. And rapidly penetrates into the pores of β-TCP, thereby realizing early cell adhesion to β-TCP. At this time, since the osteoblasts on β-TCP are in an activated state in advance, these cells try to integrate quickly in β-TCP. As a result, cell adhesion to β-TCP is realized more quickly and accurately.
以下、本発明に係る実施例について説明する。本実施例においては、実験用の兎を用いて、骨補填材を補填した後の骨形成状態を観察・確認した。 Examples according to the present invention will be described below. In this example, the bone formation state after supplementing the bone grafting material was observed and confirmed using an experimental scissors.
実験に用いた兎は、ニュージーランド白色家兎とし、ＳＰＦ（Ｓｐｅｃｉｆｉｃ ｐａｔｈｏｇｅｎ−ｆｒｅｅ）の雄で、体重は約３ｋｇのものを、９羽用意した。各々の兎の頭蓋骨頂骨に縦皮切を加え、頭頂骨を露出させた。この際、骨膜は温存した。そして、各兎の頭頂骨の、縫合栓に左右対称となる２箇所に、直径１０ｍｍの骨欠損部を各々作製し、これら骨欠損部に骨補填材を補填した後、骨膜を縫合した。なお、縫合糸はナイロン製のものを用いた。 Nine white white rabbits, SPF (Specific pathogen-free) males with a body weight of about 3 kg were prepared. A longitudinal incision was made on the skull parietal bone of each heel to expose the parietal bone. At this time, the periosteum was preserved. Then, bone defect portions having a diameter of 10 mm were respectively produced at two locations on the parietal bone of each heel that were symmetrical with respect to the suture plugs, and after filling the bone defect material with these bone defect portions, the periosteum was sutured. The suture was made of nylon.
骨補填材として用いた材料は、次の通りである。
β−ＴＣＰ： ０．１５ｇ（０．２ｍｌ）
骨髄 ： ０．２ｍｌ
ＰＲＰ ： ０．２ｍｌ
β−ＴＣＰは、多孔質状の顆粒であって、気孔率７５％、その直径は０．５〜１．５ｍｍのものを用いた。このβ−ＴＣＰを、上記同様に、補填される骨欠損部の形状に合致する形状として、予めブロック状に成形した。また骨髄は、各々の兎の大腿骨顆部より採取したものを用いた。更にＰＲＰは、各々の兎から血液を採取し、この血液から抽出・作製したものを用いた。
The materials used as the bone grafting material are as follows.
β-TCP: 0.15 g (0.2 ml)
Bone marrow: 0.2ml
PRP: 0.2ml
β-TCP is a porous granule having a porosity of 75% and a diameter of 0.5 to 1.5 mm. In the same manner as above, this β-TCP was previously formed into a block shape so as to match the shape of the bone defect to be compensated. Bone marrow was collected from the femoral condyle of each heel. Furthermore, PRP used blood which extract | collected from each sputum, extracted and produced from this blood.
ここでは９羽の兎を、補填した骨補填材の種類によって、Ａグループ（６羽）とＢグループ（３羽）の２つに分類した。
Ａグループに属する６羽には、上記２箇所の骨欠損部のうちの一方に、ＰＲＰ及び骨髄を含有させたβ−ＴＣＰを、他方に、ＰＲＰを含有させたβ−ＴＣＰを、各々骨補填材として補填した。またＢグル−プに属する３羽には、上記２箇所の骨欠損部の双方に、β−ＴＣＰのみを骨補填材として各々補填した。なお、各兎に対して、感染防止のための抗生剤の使用は、状況に応じて適宜可能とした。
Here, nine wings were classified into two groups, A group (6 wings) and B group (3 wings), depending on the type of bone prosthesis that was supplemented.
Six birds belonging to Group A were bone-supplemented with β-TCP containing PRP and bone marrow in one of the two bone defects, and β-TCP containing PRP on the other. Supplemented as a material. In addition, β-TCP alone was supplemented as a bone grafting material to both of the two bone defects in the three birds belonging to the B group. It should be noted that the use of antibiotics for preventing infection was appropriately made possible for each sputum depending on the situation.
骨補填材の補填後、所定期間経過時に屠殺し、各々の頭頂部を摘出し中性ホルマリンにて固定した後、患部を観察するとともに、顕微鏡写真を各々撮影した。所定期間経過時とは、２週間、３週間及び４週間が各々経過した時点とした。各時点において、Ａグループから２羽ずつ、Ｂグループから１羽ずつを観察対象とした。 After supplementing the bone grafting material, it was sacrificed at the elapse of a predetermined period, and each parietal part was removed and fixed with neutral formalin. Then, the affected part was observed and micrographs were taken. The time when the predetermined period passed was the time when 2 weeks, 3 weeks and 4 weeks passed. At each time point, two birds from group A and one bird from group B were observed.
所定期間経過後屠殺し、各々の頭頂部を摘出し中性ホルマリンにて固定した後、患部を観察するとともに、顕微鏡写真を各々撮影した。これら写真を図１乃至図３に各々示す。図１には２週間経過時（２Ｗ経過時）を、図２には３週間経過時（３Ｗ経過時）を、図３には４週間経過時（４Ｗ経過時）を、各々示している。各図において、（ａ）はβ−ＴＣＰのみを骨補填材として用いた場合（ＴＣＰ単独）、（ｂ）はＰＲＰを含有するβ−ＴＣＰを骨補填材として用いた場合（ＰＲＰ複合ＴＣＰ）、（ｃ）はＰＲＰ及び骨髄を含有するβ−ＴＣＰを骨補填材として用いた場合（ＰＲＰ／骨髄複合ＴＣＰ）である。 After a predetermined period of time, the mice were sacrificed, and the tops of the heads were extracted and fixed with neutral formalin, and then the affected areas were observed and micrographs were taken. These photographs are shown in FIGS. 1 to 3, respectively. FIG. 1 shows the time when 2 weeks have passed (when 2 W has passed), FIG. 2 shows the time when 3 weeks have passed (when 3 W has passed), and FIG. 3 shows the time when 4 weeks have passed (when 4 W has passed). In each figure, (a) is the case where only β-TCP is used as a bone prosthetic material (TCP alone), (b) is the case where β-TCP containing PRP is used as a bone prosthetic material (PRP composite TCP), (C) is the case where β-TCP containing PRP and bone marrow is used as a bone grafting material (PRP / bone marrow composite TCP).
これらの写真から、ＴＣＰ単独よりもＰＲＰ複合ＴＣＰの方が、またＰＲＰ複合ＴＣＰよりもＰＲＰ／骨髄複合ＴＣＰの方が、より骨形成が促進され、自家骨への置換がより短期間で的確に行われていることが明らかである。 From these photographs, the PRP composite TCP is more accelerated than the TCP alone, and the PRP / bone marrow composite TCP is more accelerated than the PRP composite TCP, and bone replacement is promoted more accurately in a shorter period of time. It is clear that this is happening.
本実施形態に係る骨補填材の製造方法においては、患者の血液を採取しＰＲＰを抽出する抽出過程と、このＰＲＰを足場材としてのβ−ＴＣＰに混合する混合過程と、を有するようにして、β−ＴＣＰにＰＲＰを含有した骨補填材を製造するようにしている。このように、患者の血液から抽出したＰＲＰをβ−ＴＣＰに混合し含有させるようにしているので、ＰＲＰに含有されている天然の形成因子によって、骨芽細胞の活性化が促進される。そのため、補填後の骨補填材において細胞接着が早期に実現されて骨形成が促進されるので、自家骨への置換を短期間で行うことができる。 In the method for manufacturing a bone grafting material according to the present embodiment, an extraction process for collecting blood from a patient and extracting PRP, and a mixing process for mixing the PRP with β-TCP as a scaffolding material are provided. In addition, a bone grafting material containing PRP in β-TCP is manufactured. Thus, since PRP extracted from the blood of a patient is mixed and contained in β-TCP, activation of osteoblasts is promoted by natural formation factors contained in PRP. Therefore, since cell adhesion is realized at an early stage and bone formation is promoted in the bone filling material after filling, replacement with autologous bone can be performed in a short period of time.
また、抽出過程の前に、患者から骨髄を採取する骨髄採取過程と、この骨髄をβ−ＴＣＰにに含浸させる含浸過程と、β−ＴＣＰに含浸された骨髄を所定時間培養する培養過程と、を有するようにして、β−ＴＣＰにＰＲＰ及び骨髄を含有した骨補填材を製造するようにしている。このように、患者から採取した骨髄もβ−ＴＣＰに含浸させるようにしているので、骨髄に含有されている骨芽細胞がβ−ＴＣＰに播種され、予め骨芽細胞を播種した状態の骨補填材とすることができる。そして、この骨芽細胞のβ−ＴＣＰ上での挙動が、ＰＲＰの形成因子の作用によって、ｉｎ−ｖｉｔｒｏの状態で活性化される。このように、骨芽細胞が活性化された状態となっている骨補填材を補填することができるので、細胞接着が更に早期に、更に的確に実現され、自家骨への置換をより短期間で行うことができる。 Further, before the extraction process, a bone marrow collection process for collecting bone marrow from a patient, an impregnation process for impregnating the bone marrow with β-TCP, a culture process for culturing the bone marrow impregnated with β-TCP for a predetermined time, Therefore, a bone filling material containing PRP and bone marrow in β-TCP is manufactured. In this way, since the bone marrow collected from the patient is also impregnated with β-TCP, osteoblasts contained in the bone marrow are seeded on β-TCP, and bone replacement in a state in which the osteoblasts are seeded in advance. It can be a material. And the behavior of this osteoblast on β-TCP is activated in an in-vitro state by the action of the formation factor of PRP. As described above, since the bone grafting material in which the osteoblasts are activated can be supplemented, cell adhesion can be realized more quickly and more accurately, and replacement with autologous bone can be performed in a shorter period of time. Can be done.
更に、培養過程において、骨髄に超音波刺激を与えるようにすれば、β−ＴＣＰと骨髄との初期親和性を高めることができ、細胞接着時間をより短縮化することができる。 Furthermore, if ultrasonic stimulation is applied to the bone marrow during the culturing process, the initial affinity between β-TCP and the bone marrow can be increased, and the cell adhesion time can be further shortened.
更に、患者から多血小板血漿を抽出する、あるいは骨髄を採取するようにしているので、一連の作業を全て骨補填手術の最中において行うことができ、手術中の短時間内に骨補填材の製造を完了し、その後速やかに患者の体内に埋め込み補填することができる。そして、多血小板血漿あるいは骨髄とも、患者から採取したものをまた同一患者の体内に戻すために、拒絶反応等を起こすおそれが無く、極めて信頼性の高い手術を的確に行うことができる。 Furthermore, since platelet-rich plasma is extracted from the patient or the bone marrow is collected, a series of operations can be performed during the bone repair operation, and within a short time during the operation, The manufacture can be completed and then immediately implanted into the patient's body. Since both platelet-rich plasma and bone marrow are collected from the patient and returned to the body of the same patient, there is no risk of rejection and the like, and an extremely reliable operation can be performed accurately.
Claims (5)
患者の血液から抽出した多血小板血漿が、前記リン酸カルシウムに含有されていることを特徴とする骨補填材。 A bone filling material using calcium phosphate as a scaffold,
A bone grafting material, wherein platelet-rich plasma extracted from the blood of a patient is contained in the calcium phosphate.
患者の血液を採取し多血小板血漿を抽出する抽出過程と、
該多血小板血漿を前記リン酸カルシウムに混合する混合過程と、
を有することを特徴とする骨補填材の製造方法。 A method for producing a bone grafting material using calcium phosphate as a scaffold,
An extraction process to collect the patient's blood and extract platelet-rich plasma;
Mixing the platelet-rich plasma with the calcium phosphate;
A method for producing a bone grafting material, comprising:
前記患者から骨髄を採取する骨髄採取過程と、
該骨髄を前記リン酸カルシウムに含浸させる含浸過程と、
前記リン酸カルシウムに含浸された前記骨髄を所定時間培養する培養過程と、
を有することを特徴とする請求項３に記載の骨補填材の製造方法。 Before the extraction process,
A bone marrow collection process for collecting bone marrow from the patient;
Impregnation step of impregnating the bone marrow with the calcium phosphate;
A culture process of culturing the bone marrow impregnated with the calcium phosphate for a predetermined time;
The method for producing a bone grafting material according to claim 3, comprising:
The method for producing a bone grafting material according to claim 4, wherein ultrasonic stimulation is applied to the bone marrow in the culturing process.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003344710A JP2005110710A (en) | 2003-10-02 | 2003-10-02 | Bone replacing material and its manufacturing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003344710A JP2005110710A (en) | 2003-10-02 | 2003-10-02 | Bone replacing material and its manufacturing method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2005110710A true JP2005110710A (en) | 2005-04-28 |
Family
ID=34538256
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003344710A Pending JP2005110710A (en) | 2003-10-02 | 2003-10-02 | Bone replacing material and its manufacturing method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2005110710A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011048803A1 (en) * | 2009-10-20 | 2011-04-28 | 日東電工株式会社 | Material for induction of hard tissue regeneration |
| KR101187555B1 (en) | 2011-05-12 | 2012-10-02 | 정문환 | Preparation method of osseous coagulum for osteoinduction |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002040071A1 (en) * | 2000-11-14 | 2002-05-23 | Osteogenesis Co., Ltd. | Compositions for forming bone or periodontium and injections for forming bone or periodontium |
-
2003
- 2003-10-02 JP JP2003344710A patent/JP2005110710A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002040071A1 (en) * | 2000-11-14 | 2002-05-23 | Osteogenesis Co., Ltd. | Compositions for forming bone or periodontium and injections for forming bone or periodontium |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011048803A1 (en) * | 2009-10-20 | 2011-04-28 | 日東電工株式会社 | Material for induction of hard tissue regeneration |
| CN102665775A (en) * | 2009-10-20 | 2012-09-12 | 日东电工株式会社 | Materials that induce hard tissue regeneration |
| JPWO2011048803A1 (en) * | 2009-10-20 | 2013-03-07 | 日東電工株式会社 | Materials for guiding hard tissue regeneration |
| US9040070B2 (en) | 2009-10-20 | 2015-05-26 | Nitto Denko Corporation | Material for induction of hard tissue regeneration |
| KR101187555B1 (en) | 2011-05-12 | 2012-10-02 | 정문환 | Preparation method of osseous coagulum for osteoinduction |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10751185B2 (en) | Treatment of skeletal voids with implantable substrate hydrated with bone marrow concentrate | |
| US8702808B2 (en) | Resorbable scaffolds for bone repair and long bone tissue engineering | |
| EP2879619B1 (en) | Bone regeneration material | |
| CN104159621B (en) | Composite device that combines porous metal and bone stimuli | |
| US20180008747A1 (en) | Bone tissue engineering by ex vivo stem cells ongrowth into three-dimensional trabecular metal | |
| US20050159820A1 (en) | Member for regenerating joint cartilage and process for producing the same, method of regenerating joint cartilage and artificial cartilage for transplantation | |
| JP4504418B2 (en) | Method of manufacturing bioactive prosthetic device for bone tissue regeneration and prosthetic device | |
| JPH10500343A (en) | Biocompatible materials and bone implants for bone repair and replacement | |
| Murata et al. | Autograft of dentin materials for bone regeneration | |
| US20120245703A1 (en) | Composite bone material implant and method | |
| JP2004008437A (en) | Cultural bone | |
| US20090298173A1 (en) | Method of preparing cell for bone tissue formation and application of cell for bone tissue formation | |
| US20210244513A1 (en) | Device and method for treatment of an artificial bone implant with blood | |
| JP2020505111A (en) | Bone regeneration material | |
| US10183095B2 (en) | Treatment of skeletal voids with implantable substrate hydrated with bone marrow concentrate | |
| JP2005110710A (en) | Bone replacing material and its manufacturing method | |
| CN102085123B (en) | Device of titanium metal wire net cage compounded with titanium metal wire engineering bone and preparation method thereof | |
| US20070010892A1 (en) | Structural body constituted of biocompatible material impregnated with fine bone dust and process for producing the same | |
| Bulgin et al. | Autologous bone marrow-derived mononuclear cells combined with β-TCP for maxillary bone augmentation in implantation procedures | |
| JP2005110709A (en) | Biotissue filler | |
| Bulgin et al. | Autologous bone marrow-derived mononuclear cells combined with β-TCP for maxillary bone augmentation in implantation procedures | |
| JP4549016B2 (en) | Biomaterial | |
| JP2004016288A (en) | Organism tissue prosthesis, method for manufacturing the same, and medical operation method | |
| JP4388260B2 (en) | Articular cartilage regeneration member | |
| JP2005111129A (en) | Bone replacement material and its manufacturing method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20060823 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20100309 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20100510 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20100511 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20101005 |