JPH01314571A - Composite artificial bone material - Google Patents
Composite artificial bone materialInfo
- Publication number
- JPH01314571A JPH01314571A JP63145625A JP14562588A JPH01314571A JP H01314571 A JPH01314571 A JP H01314571A JP 63145625 A JP63145625 A JP 63145625A JP 14562588 A JP14562588 A JP 14562588A JP H01314571 A JPH01314571 A JP H01314571A
- Authority
- JP
- Japan
- Prior art keywords
- cbf
- bone
- pores
- composite artificial
- artificial bone
- 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 42
- 239000002131 composite material Substances 0.000 title claims abstract description 26
- 239000000463 material Substances 0.000 title claims abstract description 22
- 239000011148 porous material Substances 0.000 claims abstract description 24
- 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 20
- 239000001506 calcium phosphate Substances 0.000 claims abstract description 18
- 239000000560 biocompatible material Substances 0.000 claims abstract description 13
- 229910000389 calcium phosphate Inorganic materials 0.000 claims abstract description 11
- 235000011010 calcium phosphates Nutrition 0.000 claims abstract description 11
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims abstract description 11
- 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 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 230000001054 cortical effect Effects 0.000 claims abstract description 8
- 229910000391 tricalcium phosphate Inorganic materials 0.000 claims abstract description 7
- 229940078499 tricalcium phosphate Drugs 0.000 claims abstract description 7
- 239000000203 mixture Substances 0.000 claims abstract description 6
- 239000000126 substance Substances 0.000 claims abstract description 6
- 235000019731 tricalcium phosphate Nutrition 0.000 claims abstract description 6
- 230000002378 acidificating effect Effects 0.000 claims abstract description 5
- 229960000789 guanidine hydrochloride Drugs 0.000 abstract description 5
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 abstract description 5
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 3
- 238000002513 implantation Methods 0.000 abstract description 3
- 238000004891 communication Methods 0.000 abstract description 2
- 239000002253 acid Substances 0.000 abstract 1
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 239000008280 blood Substances 0.000 abstract 1
- 210000004369 blood Anatomy 0.000 abstract 1
- 230000001939 inductive effect Effects 0.000 abstract 1
- 230000011164 ossification Effects 0.000 description 7
- VSIIXMUUUJUKCM-UHFFFAOYSA-D pentacalcium;fluoride;triphosphate Chemical compound [F-].[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 VSIIXMUUUJUKCM-UHFFFAOYSA-D 0.000 description 7
- 229910052586 apatite Inorganic materials 0.000 description 6
- 230000007547 defect Effects 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000002188 osteogenic effect Effects 0.000 description 4
- 230000002138 osteoinductive effect Effects 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102100026632 Mimecan Human genes 0.000 description 3
- 101800002327 Osteoinductive factor Proteins 0.000 description 3
- 210000000963 osteoblast Anatomy 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- QHRCVGXBBIRPTE-UHFFFAOYSA-N 5-ethyl-4-(4-methoxyphenyl)-1,3-thiazol-2-amine Chemical compound S1C(N)=NC(C=2C=CC(OC)=CC=2)=C1CC QHRCVGXBBIRPTE-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 102100037313 Core-binding factor subunit beta Human genes 0.000 description 2
- 101710145030 Core-binding factor subunit beta Proteins 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 235000018906 Bauhinia malabarica Nutrition 0.000 description 1
- 244000300022 Bauhinia malabarica Species 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 229910001069 Ti alloy Inorganic materials 0.000 description 1
- 241001447056 Uristes Species 0.000 description 1
- 239000005312 bioglass Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 239000002966 varnish Substances 0.000 description 1
Landscapes
- Materials For Medical Uses (AREA)
Abstract
Description
【発明の詳細な説明】
産業上の利用分野
本発明は、骨形成能に優れた複合人工骨材料に関するも
のである。さらに詳しくいえば本発明は、骨誘導因子で
ある皮質骨因子（以下ＣＢＦという）と生体親和性材料
から成る複合人工骨材料に関するものである。DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a composite artificial bone material with excellent osteogenic ability. More specifically, the present invention relates to a composite artificial bone material comprising cortical bone factor (hereinafter referred to as CBF), which is an osteoinductive factor, and a biocompatible material.
従来の技術
生体親和性材料、例えば水酸アパタイトやリン酸三カル
シウムなどのリン酸カルシウム系化合物等は、毒性がな
く、生体内において骨と結合しやすいので、人工骨や人
工歯根のような生体硬組織代替材料として利用されてい
る。Conventional technology Biocompatible materials, such as calcium phosphate compounds such as hydroxyapatite and tricalcium phosphate, are non-toxic and easily integrate with bone in living organisms, so they can be used in living hard tissues such as artificial bones and artificial tooth roots. It is used as an alternative material.
しかしながら、このような生体親和性材料は、骨伝導能
はあっても骨誘導能はないため、これを人工骨として骨
膜欠損している広範囲な骨欠損部や放射線照射を受けた
骨欠損部などへの移植には用いることができない。However, such biocompatible materials have osteoconductivity but do not have osteoinductive properties, so they can be used as artificial bones in areas with extensive bone defects that have periosteal defects or bone defects that have been exposed to radiation. It cannot be used for porting to.
他方、ユリスト（［Ｉｒ１ｓｆ）らは、骨を塩酸等で脱
灰して得た非コラーゲン性タンパク質をＢＭＰと命名し
、これが骨誘導能を有することを見出している。On the other hand, Juristo et al. ([Ir1sf) named a non-collagen protein obtained by decalcifying bone with hydrochloric acid or the like as BMP, and found that it has osteoinductive ability.
しかしながら、未だこのＢＭＰのタンパクの単体分離は
行われておらず、その構造や性質については明らかにさ
れていない。However, this BMP protein has not yet been isolated as a single protein, and its structure and properties have not been clarified.
発明が解決しようとする課題
本発明は、前記した従来の生体親和性材料がもつ骨形成
能上の欠点を克服し、移植後短期間で骨形成を可能とす
る複合人工骨材料を提供することを目的としてなされＩ
こものである。Problems to be Solved by the Invention An object of the present invention is to provide a composite artificial bone material that overcomes the drawbacks of the conventional biocompatible materials described above in terms of bone formation ability and enables bone formation in a short period of time after implantation. Made for the purpose of
It's a small thing.
課題を解決するための手段
本発明者らは、優れた骨形成能を有する複合人工骨材料
を得るために鋭意研究を重ねｔ；結果、生体親和性材料
に骨誘導因子のＣＢＦを加えた複合材がその目的に適合
しうろことを見出し、この知見に基づいて本発明をなす
に至った。Means for Solving the Problems The present inventors have conducted extensive research in order to obtain a composite artificial bone material with excellent osteogenic ability; as a result, we have developed a composite material in which the osteoinductive factor CBF is added to a biocompatible material. It was discovered that the material is suitable for the purpose, and based on this knowledge, the present invention was made.
すなわち、本発明は、分子量１４，０００〜Ｈ，００Ｇ
。That is, the present invention has a molecular weight of 14,000 to H,00G.
.
水に不溶性、弱酸性物質に可溶性の粉末状皮質骨因子と
生体親和性材料から成る複合人工骨材料を提供するもの
である。The present invention provides a composite artificial bone material consisting of a powdered cortical bone factor that is insoluble in water and soluble in a weakly acidic substance and a biocompatible material.
この複合人工骨材料は、生体親和性材料に対し、少なく
とも０．０５重量％以上の粉末状皮質骨因子（以下ＣＢ
Ｆと略記）を加え、混合、付着することにより得られる
。ＣＢＦは微量でも高効果を有し、その添加量は通常０
．１〜１０重量％程度であり、好ましくは０．２〜２重
量％である。This composite artificial bone material contains powdered cortical bone factor (hereinafter referred to as CB) of at least 0.05% by weight based on the biocompatible material.
(abbreviated as F), mixed, and attached. CBF has a high effect even in a small amount, and the amount added is usually 0.
．． The amount is about 1 to 10% by weight, preferably 0.2 to 2% by weight.
この際、原料として用いる生体親和性材料としては、例
えばリン酸カルシウム系焼結体、バイオガラス、チタン
系合金などが挙げられ、これらの中で特に、リン酸カル
シウム系焼結体が好ましい。At this time, examples of biocompatible materials used as raw materials include calcium phosphate sintered bodies, bioglass, titanium alloys, and the like, and among these, calcium phosphate sintered bodies are particularly preferred.
このリン酸カルシウム系焼結体としては、例えば水酸ア
パタイト、７ツ化アパタイト、塩化アパタイト、リン酸
三カルンウムなどの焼結体などがある。特に水酸アパタ
イトは骨生成効果の点で好ましい。これら焼結体の原料
リン酸カルシウム系化合物は各種を推動物の骨、歯から
回収された生体アパタイトのような天然産のものでもよ
いし、また湿式法や乾式法で製造されたリン酸カルシウ
ムや合成アパタイトのような合成品でもよい。これらは
１〜１０００μｍ程度の粉末又は０．１〜３Ｉ＋ＩＷ程
度のか粒状で用いられる。Examples of the calcium phosphate-based sintered body include sintered bodies of hydroxyapatite, heptadated apatite, chlorinated apatite, tricarium phosphate, and the like. In particular, hydroxyapatite is preferable in terms of its osteogenic effect. The raw material calcium phosphate compounds for these sintered bodies may be naturally occurring ones, such as biological apatite recovered from the bones and teeth of various animals, or calcium phosphate produced by wet or dry methods, or synthetic apatite. Synthetic products such as these may also be used. These are used in the form of powders of about 1 to 1000 μm or granules of about 0.1 to 3I+IW.
このリン酸カルシウム系焼結体としては、多孔質のもの
が好ましく、このようなものとしては、通常孔径１０〜
２００２層の気孔及び２０〜５０％の気孔率を有するも
のが一般的であり、この中で特に気孔が孔径１０＝１０
０１１ｍの細孔と孔径１００〜２００μｍの孔からなる
ものが好ましい。This calcium phosphate-based sintered body is preferably porous, and such a body usually has a pore size of 10 to 10.
It is common to have 2002 layers of pores and a porosity of 20 to 50%.
Preferably, the material has 0.011 m pores and pores with a pore diameter of 100 to 200 μm.
この１０−１００μ属の細孔は骨形成に適している上、
１００ｐＨ１より大きい孔に比べてＣＢＦの担持面積を
大きくしてＣＢＦの活性を十分に発揮させることができ
、この点からも骨形成に役立つ。また、１００〜２００
μｍの大きい孔は骨形成用の養分等を運ぶ毛細血管のた
めの連通孔として有用である。These 10-100μ pores are suitable for bone formation, and
Compared to pores with a pH of more than 100, the CBF supporting area can be increased and the CBF activity can be fully exerted, which is also useful for bone formation. Also, 100-200
The large pores of μm are useful as communication pores for capillaries that carry nutrients for bone formation.
上記リン酸カルシウム系焼結体として好適には、多孔質
水酸アパタイトとリン酸三カルシウムとの混合体、例え
ば多孔質水酸アパタイト１００重量部とσ−リン酸三カ
ルシウムｌＯ〜１００重量部との混合物に水を加え７０
〜１００°Ｃにおいて硬化させたものを用いることがで
きる。この場合、さらに加熱処理をしてβ−リン酸三カ
ルシウムに転化させて得た複合焼結体材料として用いる
こともできる。The calcium phosphate-based sintered body is preferably a mixture of porous hydroxyapatite and tricalcium phosphate, such as a mixture of 100 parts by weight of porous hydroxyapatite and 100 parts by weight of σ-tricalcium phosphate. Add water to 70
Those cured at ~100°C can be used. In this case, it can also be used as a composite sintered material obtained by further heat treatment to convert it into β-tricalcium phosphate.
この複合材料は大きな欠損部に用いることができ、取扱
いやすいので好ましい。This composite material is preferred because it can be used for large defects and is easy to handle.
この複合材料における多孔質水酸アパタイトとリン酸三
カルシウムとの配合比は、通常、重量比でｌｏ：ｌ〜１
：１０の範囲のものであり、ｌニア〜７：１特に１：１
〜７：ｌが好ましい。これよりも多孔質水酸アパタイト
の量が多くなると気孔率か高くなりすぎて強度が不十分
となるし、また、これよりも少なくなると気孔率が低下
し多孔質アパタイトの望ましい性質、例えば複合人工骨
材としての生体同化能が低下するのを免れない。The blending ratio of porous hydroxyapatite and tricalcium phosphate in this composite material is usually lo:l to 1 by weight.
:10 range, from 1:1 to 7:1 especially 1:1
~7:l is preferred. If the amount of porous hydroxyapatite is greater than this, the porosity will be too high and the strength will be insufficient, and if it is less than this, the porosity will decrease and the desirable properties of porous apatite, such as composite artificial It is inevitable that the bioassimilation ability as an aggregate will decrease.
次に他方の原料として用いるＣＢＦは、粉末状で、水に
不溶性、弱酸性物質に可溶性、分子量１４，０００〜Ｉ
Ｉ、Ｇｏｏの主としてタンパク質系の骨誘導因子であり
、そのアミノ酸連鎖構造はＮ−末端配列として、Ｔｙｒ
−Ｌｃｍ−（Ａｓｐ又は５ｅｔ）−（Ｔｙｒ又はＡｌｔ
）−（Ｇｌ＋＋又はＡｓｐ）−Ｌｃｍ−Ｌｃ＋＋−Ａｌ
ｔ−Ａｓｐ−（Ｖａｔ又はＴｙｒ）−配列やＴｙｒ−Ｌ
ｓ＋＋−（Ａｓｐ又は５ｅｔ）−Ａｌａ−Ｇｉｎ−ＬＣ
ｕ−（Ｌｅ＋＋又はＧＩＹ）−＾ｌ５−Ａ５ｐ−Ｙｘｌ
−配列（ここで本印部は未確定である）を含有するもの
である。Ｎ−末端配列が前者のものは分子ｆｆ１１４．
Ｇｏｏ −Ｉｓ、Ｏｎであり（以下このものをＣＢＦ−
αという）、Ｎ−末端配列が後者のものは分子量ＩＴ、
０００〜１１，０００である（以下このものをＣＢＦ−
βという）。Next, CBF used as the other raw material is in powder form, insoluble in water, soluble in weakly acidic substances, and has a molecular weight of 14,000 to I
I, Goo is mainly a protein-based osteoinductive factor, and its amino acid chain structure is Tyr as an N-terminal sequence.
-Lcm-(Asp or 5et)-(Tyr or Alt
)-(Gl++ or Asp)-Lcm-Lc++-Al
t-Asp-(Vat or Tyr)-sequence or Tyr-L
s++-(Asp or 5et)-Ala-Gin-LC
u-(Le++ or GIY)-^l5-A5p-Yxl
- Contains an array (here, the main mark is undefined). The former N-terminal sequence is the molecule ff114.
Goo -Is, On (hereinafter referred to as CBF-
α), the latter N-terminal sequence has a molecular weight of IT,
000 to 11,000 (hereinafter referred to as CBF-
).
ＣＢＦを溶解する弱酸性物質としては、例えば希塩酸、
酢酸、ギ酸などが挙げられ、特に塩酸グアニジンが好ま
しい。Examples of weakly acidic substances that dissolve CBF include dilute hydrochloric acid,
Examples include acetic acid and formic acid, with guanidine hydrochloride being particularly preferred.
このＣＢＦを製造するには、例えば哺乳類特に牛の長骨
の皮質骨のみを凍結粉砕し、希塩酸で脱灰し、脱脂し、
塩酸グアニジン溶液を用いて溶解抽出し、遠心分離し、
沈殿を除去した残留物を塩酸グアニジン溶液を用いて溶
解し、水で透析し、沈殿物を凍結乾燥し、ゲルろ過によ
り分画精製し、水で透析し、遠心分離し、次いで沈殿物
を凍結乾燥するなどの手段が取られる。ゲルろ過に際し
ては、セファクリルニス（Ｓｅｐｂａｃｒｙｌ Ｓ）
−２００゜セファデックス ジーＣ３ｅｐｂｓｄｅｘ
Ｇ） −７５カラムなどが用いられる。To produce this CBF, for example, only the cortical bone of the long bone of a mammal, especially a cow, is freeze-pulverized, demineralized with dilute hydrochloric acid, defatted,
Dissolve and extract using guanidine hydrochloride solution, centrifuge,
The residue from which the precipitate was removed is dissolved using guanidine hydrochloride solution, dialyzed against water, the precipitate is freeze-dried, fractionated and purified by gel filtration, dialyzed against water, centrifuged, and then the precipitate is frozen. Measures such as drying are taken. For gel filtration, use Sephacryl varnish (Sepbacryl S)
-200゜Sephadex G C3epbsdex
G) -75 column etc. are used.
さらに具体的方法について述べると、上記分画精製物の
中で最も骨誘導活性の強いフラクションをラエムリ（Ｌ
ｓｅｍｍｌｉ）の方法に準じたＳＤＳ −ＰＡＧＥでさ
らに分離し、セミトライブロッティング（Ｓ ｅｍｉｄ
ｒｙｂｌｏ口ｉｎＯ装置〔サルトリアス（社）製〕を用
いてイモピロン（ＰＶＤＦ）又はポリプレンコートグラ
スフィルターに転写し、次いでこれをプロテインシーク
エンサー（Ｐｒｏｔｓｉｎ ５ｅｑａｅ＋＋ｃｅｒ）
（アプライドバイオシステム（社）製Ｍｏｄｅｌ ４７
７Ａ）を用いてＳＤＳ −ＰＡＧＥで確認された主たる
バンドについてＮ末端付近のアミノ酸配列を分析した。More specifically, the fraction with the strongest osteoinductive activity among the above fractionated and purified products was extracted from Laemli (L).
Further separation was performed by SDS-PAGE according to the method of Semmli, and semitribe blotting (Semid
It was transferred to imopilone (PVDF) or a polyprene coated glass filter using a ryblo mouth-in-O device (manufactured by Sartorius), and then transferred to a protein sequencer (Protsin 5eqae++cer).
(Model 47 manufactured by Applied Biosystems)
7A), the amino acid sequence near the N-terminus of the main band confirmed by SDS-PAGE was analyzed.
なお、分子量測定はラエムリ（Ｌａｅ＋ａｍｌｉ）の方
法に準じ、５ＤＳ−ポリアクリルアミドゲル電気泳動法
により行っｔ；。The molecular weight was measured by 5DS-polyacrylamide gel electrophoresis according to the method of Lae + Amli.
発明の効果
本発明の複合人工骨材料は、骨形成能に優れ、比較例か
ら明らかなように既知のＢＭＰに比べ少量のＣＢＦでも
有効である上に、従来大きな骨欠損部へブロック人工骨
を移植すると、繊維芽細胞（筋肉になる細胞）が骨芽細
胞より先に出現し、骨形成量が不十分となる傾向があっ
たが、ＣＢＦを用いたことにより骨芽細胞が先に出現し
てこのような不都合を生じないという顕著な効果を奏す
る。Effects of the Invention The composite artificial bone material of the present invention has excellent osteogenic ability, and as is clear from comparative examples, it is effective even with a small amount of CBF compared to known BMP, and it is also difficult to apply block artificial bone to large bone defects. When transplanted, fibroblasts (cells that become muscle) appeared before osteoblasts, and the amount of bone formation tended to be insufficient; however, by using CBF, osteoblasts appeared first. This has the remarkable effect of not causing such inconvenience as leverage.
したがって、本発明の複合人工骨材料は、整形外科、口
腔外科、歯科等の分野での治療用の人工骨、人工歯根の
材料として好適である。Therefore, the composite artificial bone material of the present invention is suitable as a material for artificial bones and artificial tooth roots for treatment in fields such as orthopedics, oral surgery, and dentistry.
実施例 次に実施例により本発明をさらに詳細に説明する。Example Next, the present invention will be explained in more detail with reference to Examples.
調製例
成牛長管骨を粉砕し脱灰及び脱脂を行った後、６Ｍ塩酸
グアニジンで抽出した。抽出物をセファクリルニス（Ｓ
ｅｐｈｉｃｒｙｌ Ｓ） −２００（７ア／Ｌ／ ？
シア（社）製）によるゲルろ適法で分画精製し、得られ
た分画精製物の中で最も骨誘導活性の強い７ラクシヨン
をＬｘｅｍｍｌ ｉの方法に準じたＳＤＳ −ＰＡＧＥ
でさらに分離し、次いで透析希釈して得た沈殿物を真空
凍結乾燥してＣＭＦを調製した。Preparation Example Adult bovine long bones were crushed, decalcified and defatted, and then extracted with 6M guanidine hydrochloride. The extract was treated with Sephacrylnis (S
ephicryl S) -200 (7A/L/?
7-lactone, which has the strongest osteoinductive activity, was subjected to SDS-PAGE according to the method of Lxemml I.
The precipitate obtained by further separation and dilution by dialysis was then lyophilized under vacuum to prepare CMF.
実施例１
ＣＢＦ （等量のＣＢＦ−ａとＣＢＦ−β）２．０１１
１９を水０．１５ｃ（に超音波で分散させ、これをか粒
状水はアパタイト焼結体（粒径４００〜８００μｍ１平
均気孔径１００ｐ＋ｍ）１２０ｍｇに付着させた。Example 1 CBF (equal amounts of CBF-a and CBF-β) 2.011
No. 19 was dispersed in 0.15 c of water using ultrasonic waves, and the granular water was attached to 120 mg of apatite sintered body (particle size: 400 to 800 μm, average pore size: 100 p+m).
この際、減圧中で処理することにより、アパタイト焼結
体の細孔内にＣＢＦを担持させた。次いで真空凍結乾燥
して複合人工骨材を作成した。このものをマウスの大腿
部筋肉の５Ｘ５Ｘ５＋＋ｎａの大きさの部位に移植した
。２週間後に骨芽細胞が見られ、新生骨の生成が認めら
れた。At this time, by processing under reduced pressure, CBF was supported within the pores of the apatite sintered body. The mixture was then vacuum freeze-dried to create a composite artificial aggregate. This was transplanted into the thigh muscle of a mouse at a site measuring 5×5×5++ na. Two weeks later, osteoblasts were observed and new bone formation was observed.
実施例２．３
実施例１のＣＢＦＩこ代えてＣＢＦ−ａ及びＣＢＦ−β
をそれぞれ単独で用いた以外は実施例１と同様にして複
合人工骨材を作成した。これらも実施例１と同様の移植
結果が得られた。Example 2.3 CBF-a and CBF-β instead of CBFI in Example 1
A composite artificial aggregate was produced in the same manner as in Example 1 except that each of the following was used alone. The same transplantation results as in Example 1 were also obtained in these cases.
実施例４
水酸アパタイト焼結体に代えて水酸アパタイトとリン酸
三カルシウムとの等量混合焼結体を用いた以外は実施例
１と同様にして複合人工骨材を作成した。このものも実
施例１と同様の移植結果が得られた。Example 4 A composite artificial aggregate was produced in the same manner as in Example 1, except that a sintered mixture of equal amounts of hydroxyapatite and tricalcium phosphate was used instead of the sintered hydroxyapatite. The same transplantation results as in Example 1 were also obtained in this case.
比較例
ＣＢＦに代えてユリスト（Ｕｒｉｓｔ）の見出したＩＩ
ＭＰを用いた以外実施例１と同様にして得た複合人工骨
材を用いて同様な移植試験を行った。その結果ＢＭＰ
５ ｍ１以上でないと新生骨が生じなかった。また、Ｂ
ＭＰ５ｍｇ用いた場合でも３週間以上経過しないと新生
骨が生じなかった。Comparative Example II discovered by Urist in place of CBF
A similar implantation test was conducted using a composite artificial aggregate obtained in the same manner as in Example 1 except that MP was used. As a result, BMP
New bone did not occur unless it was 5 m1 or more. Also, B
Even when 5 mg of MP was used, new bone did not occur until 3 weeks or more had elapsed.
Claims (1)
弱酸性物質に可溶性の粉末状皮質骨因子と生体親和性材
料から成る複合人工骨材料。 ２ 皮質骨因子が以下に示すＮ−末端配列を有する分子
量１４，０００〜１５，０００のものである請求項１記
載の複合人工骨材料： Ｔｙｒ−Ｌｅｎ−（Ａｓｐ又はＳｅｒ）−（Ｔｙｒ又は
Ａｌａ）−（Ｇｌｕ又はＡｓｐ）−Ｌｅｎ−Ｌｅｎ−Ａ
ｌａ−。 ３ 皮質骨因子が以下に示すＮ−末端配列を有する分子
量１７，０００〜１１，０００のものである請求項１記
載の複合人工骨材料： Ｔｙｒ−Ｌｅｎ−（Ａｓｐ又はＳｅｒ）−Ａｌａ−Ｇｌ
ｕ−Ｌｅｎ（Ｌｅｎ又はＧｌｙ）−Ａｌａ−。 ４ 生体親和性材料がリン酸カルシウム系焼結体である
請求項１ないし３のいずれかに記載の複合人工骨材料。 ５ リン酸カルシウム系焼結体が孔径１０〜２００μｍ
の気孔及び２０〜５０％の気孔率を有する請求項４記載
の複合人工骨材料。 ６ 気孔が孔径１０〜１００μｍの細孔と孔径１００〜
２００μｍの孔からなる請求項５記載の複合人工骨材料
。 ７ リン酸カルシウム系焼結体が水酸アパタイトとリン
酸三カルシウムとの混合体である請求項４ないし６のい
ずれかに記載の複合人工骨材料。[Scope of Claims] 1. Molecular weight 14,000 to 18,000, insoluble in water,
A composite artificial bone material consisting of powdered cortical bone factor soluble in weakly acidic substances and a biocompatible material. 2. The composite artificial bone material according to claim 1, wherein the cortical bone factor has the following N-terminal sequence and has a molecular weight of 14,000 to 15,000: Tyr-Len-(Asp or Ser)-(Tyr or Ala )-(Glu or Asp)-Len-Len-A
la-. 3. The composite artificial bone material according to claim 1, wherein the cortical bone factor has the following N-terminal sequence and has a molecular weight of 17,000 to 11,000: Tyr-Len-(Asp or Ser)-Ala-Gl
u-Len(Len or Gly)-Ala-. 4. The composite artificial bone material according to any one of claims 1 to 3, wherein the biocompatible material is a calcium phosphate sintered body. 5 Calcium phosphate sintered body has a pore diameter of 10 to 200 μm
The composite artificial bone material according to claim 4, having pores of 20 to 50% and a porosity of 20 to 50%. 6 Pores with a pore diameter of 10 to 100 μm and pores with a pore diameter of 100 to 100 μm
The composite artificial bone material according to claim 5, comprising pores of 200 μm. 7. The composite artificial bone material according to any one of claims 4 to 6, wherein the calcium phosphate-based sintered body is a mixture of hydroxyapatite and tricalcium phosphate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63145625A JPH01314571A (en) | 1988-06-15 | 1988-06-15 | Composite artificial bone material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63145625A JPH01314571A (en) | 1988-06-15 | 1988-06-15 | Composite artificial bone material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01314571A true JPH01314571A (en) | 1989-12-19 |
Family
ID=15389340
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63145625A Pending JPH01314571A (en) | 1988-06-15 | 1988-06-15 | Composite artificial bone material |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01314571A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8603184B2 (en) | 2002-04-03 | 2013-12-10 | DePuy Synthes Products, LLC | Kneadable and pliable bone replacement material |
| CN107440913A (en) * | 2017-07-27 | 2017-12-08 | 武汉大学 | A kind of preparation method of coloured hydroxyapatite available for the detection of dentistry bleaching agent effectiveness |
-
1988
- 1988-06-15 JP JP63145625A patent/JPH01314571A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8603184B2 (en) | 2002-04-03 | 2013-12-10 | DePuy Synthes Products, LLC | Kneadable and pliable bone replacement material |
| CN107440913A (en) * | 2017-07-27 | 2017-12-08 | 武汉大学 | A kind of preparation method of coloured hydroxyapatite available for the detection of dentistry bleaching agent effectiveness |
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