【０００１】[0001]
【発明の属する技術分野】本発明は、人工血管に関し、
更に詳しくは、冠状動脈や末梢血管などの小口径血管の
代替血管として用いる人工血管に関する。TECHNICAL FIELD The present invention relates to an artificial blood vessel,
More specifically, it relates to an artificial blood vessel used as a substitute blood vessel for a small diameter blood vessel such as a coronary artery or a peripheral blood vessel.
【０００２】[0002]
【従来の技術】従来より、ポリエステル繊維の織物また
は編み物や延伸ポリテトラフルオロエチレン（以下、Ｅ
ＰＴＦＥという）のチューブが人工血管として用いられ
ている。ＥＰＴＦＥチューブは素材であるポリテトラフ
ルオロエチレン自体が抗血栓性に優れる上、延伸によっ
て得られる繊維−結節からなる多孔質構造が生体組織適
合性に優れるため、ポリエステルに比べてより小口径の
人工血管に適用されてきた。2. Description of the Related Art Conventionally, woven or knitted polyester fibers and expanded polytetrafluoroethylene (hereinafter referred to as E
A tube of PTFE) is used as an artificial blood vessel. EPTFE tubing is made of polytetrafluoroethylene, which itself has excellent antithrombogenicity, and the porous structure consisting of fibers and nodules obtained by stretching is excellent in biocompatibility, making it a smaller diameter artificial blood vessel than polyester. Has been applied to.
【０００３】しかしながらＥＰＴＦＥでも抗血栓性が十
分ではなく、内径５ｍｍ以下、特に内径４ｍｍ以下の人
工血管では十分な開存性は得られていない。そこでこれ
らを解決する方法として、材料自体の抗血栓性を高め
る方法、人工血管内に、抗血栓性の組織を培養（播
種）する方法、人工血管を移植後に、内面に抗血栓性
の組織形成を促すように工夫する方法が検討されてい
る。However, even EPTFE does not have sufficient antithrombogenicity, and sufficient patency has not been obtained with an artificial blood vessel having an inner diameter of 5 mm or less, particularly 4 mm or less. Therefore, as a method of solving these problems, a method of increasing the antithrombotic property of the material itself, a method of culturing (seeding) an antithrombotic tissue in an artificial blood vessel, and a method of forming an antithrombotic tissue on the inner surface after transplanting an artificial blood vessel The method of devising so as to encourage is being considered.
【０００４】[0004]
【発明が解決しようとする課題】具体的には、の方法
としては、血栓が付着しない合成高分子材料や、抗血栓
物質を固定化した材料の開発が検討されている（野一色
ら、トランザクションズ・オブ・アサイオ（Ｔｒａｎ
ｓ．Ａ．Ｓ．Ａ．Ｉ．Ｏ．）、２３、２５３（１９７
７）など）。これらの方法では、人工血管を移植した後
の一時的な抗血栓性の付与は可能であるが、長期間を経
ると血栓を形成して閉塞に至る。の方法としては、人
工血管内面に血管内皮細胞を播種する方法が提案されて
いるが、ヒトの血管内皮細胞の確保が困難でかつ、培養
に数週間を要することが問題であり実用化されていない
（高木ら、人工臓器、１７、６７９（１９８８）、特開
平１−１７０４６６など）。Specifically, as a method of (3), development of a synthetic polymer material to which thrombus does not adhere and a material to which an antithrombotic substance is immobilized have been studied (Noichishiro et al., Transaction). The Of Acai (Tran
s. A. S. A. I. O. ),23 , 253 (197)
7) etc.). With these methods, it is possible to temporarily impart antithrombotic properties after transplanting an artificial blood vessel, but after a long period of time, thrombus is formed and obstruction occurs. As a method of, a method of seeding vascular endothelial cells on the inner surface of an artificial blood vessel has been proposed, but it is difficult to secure human vascular endothelial cells, and it takes a few weeks for culturing, which has been put to practical use. No. (Takagi et al., Artificial Organ, 17, 679 (1988), JP-A-1-170466, etc.).
【０００５】の方法としては、内皮細胞の接着性物質
や増殖性物質を塗布または共有結合により固定した材料
が提案されている。内皮細胞接着性物質、または内皮細
胞接着性物質と増殖性物質の両方を塗布した人工血管が
提案されているが、内皮細胞の形成は促進されず開存性
も向上しない。（ルンドグレンら、トランザクションズ
・オブ・アサイオ、３２、３４６（１９８６）、グライ
スラーら、サージャリー（ＳＵＲＧＥＲＹ）、１１２、
２４４（１９９２）など）。内皮細胞接着性物質を化学
固定した人工血管が提案されているが、効果は不十分で
ある（特開平５−２６９１９８）。さらに、内皮細胞接
着性物質と増殖性物質の両方を共有結合固定した材料が
in vitroで内皮細胞の形成を促進した、との報告がある
が、人工血管として移植すると効果は少なく、開存性も
満足できるものではない（ジャーナル・オブ・バイオメ
ディカル・マテリアルズ・リサーチ（Ｊ．Ｂiomed．Ｍa
ter．Ｒes.）２７、９０１、（１９９３）など）。ま
た、人工血管を移植後、内皮細胞の形成が完了するまで
の間の血栓の形成を抑制するために、内皮細胞の接着物
質であるコラーゲンまたはゼラチンとヘパリンとを塗布
した人工血管が提案されているが、内皮細胞の形成およ
び血栓形成の抑制のいずれも不十分であり、良好な開存
性は得られていない（特開昭６３−４６１６９）。As the method (1), a material in which an adhesive substance or a proliferative substance for endothelial cells is applied or fixed by covalent bonding has been proposed. An artificial blood vessel coated with an endothelial cell adhesive substance or both an endothelial cell adhesive substance and a proliferative substance has been proposed, but the formation of endothelial cells is not promoted and patency is not improved. (Lundgren et al., Transactions of Acai, 32, 346 (1986), Greisler et al., Surgery,112 ,
244 (1992)). An artificial blood vessel in which an endothelial cell adhesive substance is chemically fixed has been proposed, but its effect is insufficient (JP-A-5-269198). Furthermore, a material in which both endothelial cell adhesive substance and proliferative substance are covalently fixed
It has been reported that it promoted the formation of endothelial cells in vitro, but when transplanted as an artificial blood vessel, the effect was small and patency was not satisfactory (Journal of Biomedical Materials Research (J. . Biomed. Ma
ter. Res.)27 , 901, (1993), etc.). Further, in order to suppress the formation of thrombus after the transplantation of the artificial blood vessel until the formation of the endothelial cells is completed, an artificial blood vessel coated with collagen or gelatin, which is an adhesive substance of the endothelial cells, and heparin has been proposed. However, neither formation of endothelial cells nor suppression of thrombus formation is insufficient, and good patency has not been obtained (JP-A-63-46169).
【０００６】[0006]
【課題を解決するための手段】本発明者らは、鋭意研究
を行った結果、ＥＰＴＦＥからなる多孔性チューブの孔
内部の壁面を含む全ての表面に生体組織誘導性物質を固
定し、要すれば抗血栓性物質を、少なくとも血液に接す
るチューブ表面に固定すれば、開存性が高まり、またＥ
ＰＴＦＥチューブの血液に接する面の繊維長を６０μｍ
以上にすれば、更に開存性が高められることを見い出
し、本発明を完成した。Means for Solving the Problems As a result of intensive studies, the present inventors have fixed a biological tissue-inducing substance on all surfaces including the wall surface inside the pores of a porous tube made of EPTFE, and required For example, if an antithrombotic substance is fixed to at least the surface of the tube that comes into contact with blood, patency is increased and
The fiber length of the surface of the PTFE tube in contact with blood is 60 μm
By the above, it was found that the patency is further enhanced, and the present invention was completed.
【０００７】即ち、本発明は、（１）延伸ポリテトラフ
ルオロエチレンの多孔質チューブからなり、該チューブ
の孔内部の壁面を含む全ての表面に生体組織誘導性物質
を固定し、かつ抗血栓性物質を少なくとも血液に接する
該チューブの表面に固定した人工血管、（２）少なくと
も血液に接する面の繊維長が６０μｍ以上である延伸ポ
リテトラフルオロエチレンの多孔質チューブからなり、
該チューブの孔内部の壁面を含む全ての表面に生体組織
誘導性物質を固定した人工血管、（３）血液に接する面
の繊維長が６０μｍ以上であり、繊維長が４０μｍ以下
である層を少なくとも一層含む多層構造を有する延伸ポ
リテトラフルオロエチレンの多孔質チューブからなり、
該チューブの孔内部の壁面を含む全ての表面に生体組織
誘導性物質を固定した人工血管、（４）少なくとも血液
に接する面の繊維長が６０μｍ以上である延伸ポリテト
ラフルオロエチレンの多孔質チューブからなり、該チュ
ーブの孔内部の壁面を含む全て表面に生体組織誘導性物
質を固定し、かつ抗血栓性物質を少なくとも血液に接す
る該チューブの表面に固定した人工血管、並びに（５）
血液に接する面の繊維長が６０μｍ以上であり、繊維長
が４０μｍ以下である層を少なくとも一層含む多層構造
を有する延伸ポリテトラフルオロエチレンの多孔質チュ
ーブであり、該チューブの孔内部の壁面を含む全て表面
に生体組織誘導性物質を固定し、かつ抗血栓性物質を少
なくとも血液に接する該チューブの表面に固定した人工
血管を提供する。That is, the present invention comprises (1) a porous tube of expanded polytetrafluoroethylene, which has a biological tissue-inducing substance fixed to all surfaces including the wall surface inside the pores of the tube, and has an antithrombotic property. An artificial blood vessel in which a substance is fixed to at least the surface of the tube in contact with blood, and (2) a porous tube of expanded polytetrafluoroethylene having a fiber length of 60 μm or more on at least the surface in contact with blood,
An artificial blood vessel in which a biological tissue-inducing substance is fixed on all surfaces including the wall surface inside the hole of the tube, (3) at least a layer having a fiber length of 60 μm or more and a fiber length of 40 μm or less on the surface in contact with blood Consisting of a porous tube of expanded polytetrafluoroethylene having a multilayer structure including one layer,
An artificial blood vessel in which a biological tissue-inducing substance is fixed on all surfaces including the wall surface inside the hole of the tube, (4) From a porous tube of expanded polytetrafluoroethylene having a fiber length of 60 μm or more on at least the surface in contact with blood An artificial blood vessel in which a biological tissue-inducing substance is fixed to all surfaces including the wall surface inside the hole of the tube, and an antithrombotic substance is fixed to at least the surface of the tube in contact with blood; and (5)
A porous tube of expanded polytetrafluoroethylene having a multilayer structure including a layer having a fiber length of 60 μm or more on the surface in contact with blood and having a fiber length of 40 μm or less, including a wall surface inside the hole of the tube Provided is an artificial blood vessel in which a biological tissue-inducing substance is fixed on the entire surface and an antithrombotic substance is fixed on at least the surface of the tube in contact with blood.
【０００８】[0008]
【発明の実施の形態】使用するＥＰＴＦＥチューブとし
ては、少なくとも血液に接触する面の繊維長が６０μｍ
以上のものが好ましく用いられる。より好ましくは、血
液に接触する面の繊維長は６０μｍ以上で、且つ繊維長
が４０μｍ以下の層を含む多層構造を有するＥＰＴＦＥ
チューブが好ましい。また壁の厚さは生体血管に合わせ
るために、３００〜１０００μｍの間で選択すれば良
い。BEST MODE FOR CARRYING OUT THE INVENTION The EPTFE tube used has a fiber length of at least 60 μm on the surface in contact with blood.
The above is preferably used. More preferably, the EPTFE having a multilayer structure including a layer having a fiber length of 60 μm or more on the surface in contact with blood and having a fiber length of 40 μm or less
Tubes are preferred. Further, the wall thickness may be selected in the range of 300 to 1000 μm in order to match the blood vessel of the living body.
【０００９】ＥＰＴＦＥチューブの多孔質体の孔内部の
壁面を含む全ての表面に生体誘導性物質を共有結合によ
り固定するには、ＥＰＴＦＥに官能基を導入する必要が
ある。このような官能基としては、カルボキシル基、水
酸基、アミノ基、エポキシ基等が好ましい。中でも、カ
ルボン酸のエステルまたは水酸基のスルホン酸エステル
に生体組織誘導性物質を反応させて、カルボン酸のエス
テルのアルコール部分、または水酸基のスルホン酸エス
テルのスルホン酸部分を置換して、生体誘導性物質を共
有結合により固定するのが好ましい。In order to covalently fix the bioinducing substance to all surfaces including the wall surface inside the pores of the porous body of the EPTFE tube, it is necessary to introduce a functional group into EPTFE. As such a functional group, a carboxyl group, a hydroxyl group, an amino group, an epoxy group and the like are preferable. Among them, by reacting a biological tissue-inducing substance with an ester of a carboxylic acid or a sulfonic acid ester of a hydroxyl group, and substituting the alcohol portion of the ester of a carboxylic acid or the sulfonic acid portion of a sulfonic acid ester of a hydroxyl group, a biological inducer Are preferably fixed by covalent bonds.
【００１０】ＥＰＴＦＥチューブの多孔質体の孔内部の
壁面を含む全ての表面に官能基を形成する方法として
は、γ線や電子線などの放射線やグロ−放電を用いても
よい。しかしながら、公知の事実より、γ線による処理
では、ＥＰＴＦＥの結晶内部深くまでＰＴＦＥが分解さ
れるためＰＴＦＥの分子量が低下し、強度が著しく低下
するため、人工血管としての実用に供するのは難しい
[モレル（Ｇ．Ｍorel）ら、ジャーナル・オブ・アプラ
イド・ポリマー・サイエンス（Ｊ．Ａppl．Ｐolym．Ｓc
i．）Ｖol．２４，７７１（１９７９）]。また、グロ−
放電処理ではＥＰＴＦＥの孔内部の壁面まで処理を施す
ことが難しく、チュ−ブの外表面または内表面のみしか
処理することができないので、多孔質ＥＰＴＦＥの孔内
部の壁面の全面にタンパク類を固定することが難しく、
人工血管の壁の中への生体組織や毛細血管の侵入を促進
させることが出来ない。Radiation such as γ-ray or electron beam or glow discharge may be used as a method of forming a functional group on all surfaces including the wall surface inside the pores of the porous body of the EPTFE tube. However, according to the known fact, in the treatment with γ-ray, since PTFE is decomposed deep inside the crystal of EPTFE, the molecular weight of PTFE is lowered and the strength is remarkably lowered, so that it is difficult to put it into practical use as an artificial blood vessel.
[G. Morel et al., Journal of Applied Polymer Science (J. Appl. Polym. Sc
i. ) Vol. 24, 771 (1979)]. In addition,
It is difficult to treat the inner wall surface of the EPTFE pores by electric discharge treatment, and only the outer or inner surface of the tube can be treated. Therefore, proteins are immobilized on the entire inner wall surface of the porous EPTFE pore. Difficult to do,
It cannot promote the invasion of living tissues and capillaries into the wall of artificial blood vessels.
【００１１】これに対し、アルカリ金属処理によれば、
ＥＰＴＦＥの内表面や外表面だけでなく、多孔質ＥＰＴ
ＦＥの孔内部の壁面の全面を均一に、表面から約数百オ
ングストロ−ムの深さ部分のみが処理される。しかもγ
線や電子線などの放射線照射のようにＥＰＴＦＥの結晶
内部深くまでＰＴＦＥを分解することがないため、ＥＰ
ＴＦＥの強度低下を招くことがない。このため、タンパ
ク質やペプチドをＥＰＴＦＥの内表面、外表面、孔内部
の壁面の全表面に共有結合によって固定することが可能
となる。従って、本発明による複合化人工血管を作製す
るためには、アルカリ金属化合物で処理することが望ま
しい。On the other hand, according to the alkali metal treatment,
Not only the inner and outer surfaces of EPTFE but also porous EPT
The entire wall surface inside the hole of the FE is uniformly processed, and only a portion having a depth of about several hundred angstroms from the surface is processed. Moreover, γ
Since the PTFE is not decomposed deeply inside the EPTFE crystal unlike the irradiation of radiation such as electron beams and electron beams, the EP
It does not lead to a decrease in TFE strength. Therefore, it becomes possible to immobilize proteins and peptides on the inner and outer surfaces of EPTFE and the entire wall surface inside the pores by covalent bonding. Therefore, it is desirable to treat with an alkali metal compound in order to produce the composite artificial blood vessel according to the present invention.
【００１２】このように、アルカリ金属化合物を用いて
ＥＰＴＦＥを脱フッ素化した後に、分子内にカルボキシ
ル基、水酸基、アミノ基、エポキシ基等の官能基を有す
る化合物を付加させて、これらの官能基を導入する。Thus, after defluorinating EPTFE using an alkali metal compound, a compound having a functional group such as a carboxyl group, a hydroxyl group, an amino group, an epoxy group in the molecule is added, and these functional groups are added. To introduce.
【００１３】アルカリ金属化合物としては、例えばメチ
ルリチウム、ｎ−ブチルリチウム、ｔ−ブチルリチウ
ム、ナトリウム−ナフタレン、ナフタレン−ベンゾフェ
ノン、ビニルリチウムなどが挙げられ、これらを溶液と
して使用する。ナトリウム−ナフタレンまたはナトリウ
ム−ベンゾフェノンは、処理によってＰＴＦＥ表面に黒
褐色の層を形成する上、均一に処理することが困難であ
るので、本発明の人工血管を作成するためにはメチルリ
チウム、ｎ−ブチルリチウム、ｔ−ブチルリチウムを用
いることが望ましい。メチルリチウム、ｎ−ブチルリチ
ウム、ｔ−ブチルリチウムはこれ自体ではフッ素を引き
抜く作用が弱いので、キレート試薬であるヘキサメチル
リン酸トリアミドやＮ,Ｎ,Ｎ',Ｎ'−テトラメチルエチ
レンジアミン等を添加することが必要である。Examples of the alkali metal compound include methyllithium, n-butyllithium, t-butyllithium, sodium-naphthalene, naphthalene-benzophenone and vinyllithium, which are used as a solution. Sodium-naphthalene or sodium-benzophenone forms a black-brown layer on the PTFE surface by treatment, and it is difficult to treat uniformly. Therefore, methyllithium and n-butyl are required for preparing the artificial blood vessel of the present invention. It is desirable to use lithium or t-butyllithium. Since methyllithium, n-butyllithium, and t-butyllithium have a weak action of extracting fluorine by themselves, hexamethylphosphoric triamide and N, N, N ', N'-tetramethylethylenediamine, which are chelating agents, are added. It is necessary to.
【００１４】分子内に水酸基、カルボキシル基、エポキ
シ基またはアミノ基を含有する物質としては、グリセロ
ール（メタ）アクリレート、２−ヒドロキシエチル（メ
タ）アクリレート、２−ヒドロキシプロピル（メタ）ア
クリレート、ポリエチレングリコール（メタ）アクリレ
ート、グリシジル（メタ）アクリレート、（メタ）アク
リル酸、アリルアミン、２−アミノエチル（メタ）アク
リレート、アクリルアミド等が挙げられる。また、無水
マレイン酸等の無水物を付加し、その後に加水分解して
もよい。As the substance containing a hydroxyl group, a carboxyl group, an epoxy group or an amino group in the molecule, glycerol (meth) acrylate, 2-hydroxyethyl (meth) acrylate, 2-hydroxypropyl (meth) acrylate, polyethylene glycol ( Examples thereof include (meth) acrylate, glycidyl (meth) acrylate, (meth) acrylic acid, allylamine, 2-aminoethyl (meth) acrylate, and acrylamide. Alternatively, an anhydride such as maleic anhydride may be added and then hydrolyzed.
【００１５】抗血栓性物質または生体組織誘導性物質を
固定するためには、単に物理的に塗布してもよいし、導
入した官能基に化学結合させてもよい。目的の物質が化
学結合によっても活性を失わない物質であれば、導入し
た官能基に化学結合させる方法を用いた方がより好まし
い。その方法はその官能基に適した方法を選択すればよ
く、好ましくは固定することによって活性を失うことが
ない方法を選択すればよい。In order to immobilize the antithrombotic substance or the biological tissue inducing substance, it may be simply physically applied or may be chemically bonded to the introduced functional group. If the target substance is a substance that does not lose its activity even by a chemical bond, it is more preferable to use a method of chemically bonding to the introduced functional group. As the method, a method suitable for the functional group may be selected, and preferably, a method which does not lose the activity by being immobilized may be selected.
【００１６】例えば水酸基、カルボキシル基およびアミ
ノ基に対しては脱水縮合により、エポキシ基に対しては
付加反応により共有結合を形成させる。水酸基に対して
は、そのままカルボジイミドを触媒として脱水縮合によ
り結合させてもよいし、水酸基に例えばトリフルオロメ
タンスルホニル基等の脱離基を導入して反応性を上げて
おいてから組織誘導性物質のアミノ基と反応させてもよ
い。またカルボキシル基に対しては、そのままカルボジ
イミド等の脱水縮合触媒を用いて結合してもよいし、Ｎ
−ヒドロキシコハク酸イミドを反応させて活性エステル
を導入して反応性を上げておいてから抗血栓物質または
組織誘導性物質のアミノ基と反応させてもよい。For example, a hydroxyl group, a carboxyl group, and an amino group are formed by dehydration condensation, and an epoxy group is formed by an addition reaction to form a covalent bond. With respect to the hydroxyl group, it may be bonded by dehydration condensation using carbodiimide as a catalyst as it is, or by introducing a leaving group such as a trifluoromethanesulfonyl group into the hydroxyl group to increase the reactivity and then to remove the tissue-inducing substance. It may be reacted with an amino group. Further, to the carboxyl group, it may be bonded as it is by using a dehydration condensation catalyst such as carbodiimide.
It is also possible to react with hydroxysuccinimide to introduce an active ester to increase the reactivity and then to react with the amino group of the antithrombotic substance or the tissue-inducing substance.
【００１７】しかし、生体組織誘導性物質を共有結合に
よって固定するための官能基としては、とりわけカルボ
ン酸のＮ−ヒドロキシこはく酸イミドエステル、または
水酸基をトリフルオロエタンスルホニル化した基が好ま
しい。すなわち、アルカリ金属を用いたグラフト重合に
よって形成したこれらの官能基に生体組織誘導性物質を
反応させ、カルボン酸のＮ−ヒドロキシこはく酸イミド
部分を置換して、または水酸基のトリフルオロエタンス
ルホニル基を置換して生体組織誘導性部分を共有結合に
よって固定すると、人工血管を移植した後の生体組織誘
導効果が大きく、優れた開存成績が得られる。However, as the functional group for immobilizing the biological tissue-inducing substance by covalent bond, an N-hydroxysuccinimide ester of carboxylic acid or a group obtained by trifluoroethanesulfonylating a hydroxyl group is particularly preferable. That is, a biological tissue-inducing substance is caused to react with these functional groups formed by graft polymerization using an alkali metal, the N-hydroxysuccinimide moiety of a carboxylic acid is substituted, or a trifluoroethanesulfonyl group of a hydroxyl group is added. By substituting and fixing the biological tissue-inducing portion by covalent bonding, the biological tissue-inducing effect after transplanting an artificial blood vessel is large, and excellent patency results are obtained.
【００１８】ＥＰＴＦＥにカルボン酸基を導入し、Ｎ−
ヒドロキシこはく酸イミドエステルとしてから生体組織
誘導性物質を共有結合により固定する方法をより具体的
に説明する。アルゴンや窒素などの不活性ガス雰囲気
下、ＥＰＴＦＥチューブをメチルリチウムのジエチルエ
ーテル溶液に浸漬しておき、ここへヘキサメチルリン酸
トリアミドを添加して０℃で３０分間放置してＰＴＦＥ
のフッ素原子を引き抜いた後、反応液を除去し、ここに
（メタ）アクリル酸のテトラヒドロフラン溶液を加えて
６０〜８０℃で１０時間反応させる。反応後に余剰の
（メタ）アクリル酸とその重合体を洗浄除去して（メ
タ）アクリル酸のグラフト体を得る。By introducing a carboxylic acid group into EPTFE, N-
A method of immobilizing a biological tissue-inducing substance by a covalent bond from hydroxysuccinimide ester will be described in more detail. The EPTFE tube was immersed in a diethyl ether solution of methyllithium under an atmosphere of an inert gas such as argon or nitrogen, hexamethylphosphoric acid triamide was added thereto, and the mixture was allowed to stand at 0 ° C for 30 minutes to obtain PTFE.
After removing the fluorine atom, the reaction solution is removed, a tetrahydrofuran solution of (meth) acrylic acid is added thereto, and the mixture is reacted at 60 to 80 ° C. for 10 hours. After the reaction, excess (meth) acrylic acid and its polymer are washed and removed to obtain a (meth) acrylic acid graft product.
【００１９】このようにして作製した（メタ）アクリル
酸グラフト化ＥＰＴＦＥチューブを、Ｎ−ヒドロキシこ
はく酸イミドとジシクロヘキシルカルボジイミドとの
１,４ージオキサン中溶液に、０℃で１２時間浸漬する
ことによって、Ｎ−ヒドロキシこはく酸イミドエステル
とする。The (meth) acrylic acid-grafted EPTFE tube thus prepared was immersed in a solution of N-hydroxysuccinimide and dicyclohexylcarbodiimide in 1,4-dioxane at 0 ° C. for 12 hours to give N -Hydroxysuccinimide ester.
【００２０】この後に、溶液のｐＨを１１とした、ＥＰ
ＴＦＥチューブを生体組織誘導性物質のリン酸緩衝溶液
に浸漬して、生体組織誘導性物質を共有結合する。After this, the pH of the solution was adjusted to 11, and EP
The TFE tube is immersed in a phosphate buffer solution of a biological tissue-inducing substance to covalently bond the biological tissue-inducing substance.
【００２１】複合化する生体組織誘導性物質としては、
細胞吸着性物質と細胞増殖性物質とが優れており、これ
ら両者を共有結合により固定することにより、最も優れ
た生体組織誘導作用を得るこことができる。細胞接着性
物質としては、例えばコラーゲン、ゼラチン、ラミニン
やフィブロネクチン等が例示できるが、特にフィブロネ
クチンが好ましい。細胞増殖性物質としては、ＴＧＦ−
α、トランスフェリン、インスリン、ＥＣＧＦ（内皮細
胞増殖因子）、ＢＰＥ（脳下垂体抽出物）、ＰＤＧＦ
（血小板由来増殖因子）、ＦＧＦ（線維芽細胞増殖因
子）等が例示できるが、中でも、ＴＧＦ−α、トランス
フェリン、インスリン、ＦＧＦなどが好ましい。The biological tissue-inducing substance to be complexed is as follows:
The cell-adsorptive substance and the cell-proliferative substance are excellent, and by immobilizing both of them by covalent bond, the most excellent biological tissue inducing action can be obtained. Examples of the cell adhesive substance include collagen, gelatin, laminin, fibronectin and the like, and fibronectin is particularly preferable. As a cell proliferative substance, TGF-
α, transferrin, insulin, ECGF (endothelial cell growth factor), BPE (pituitary extract), PDGF
(Platelet-derived growth factor), FGF (fibroblast growth factor) and the like can be exemplified, but among them, TGF-α, transferrin, insulin, FGF and the like are preferable.
【００２２】抗血栓性物質としては、ヒルジン、ヘパリ
ン等の抗凝固薬、t-ＰＡやウロキナーゼ等のプラスミノ
ーゲンアクチベータ、プラスミンやスブチリシン等の線
溶酵素、プロスタサイクリンやアスピリン等の抗血小板
剤を用いればよいが、ヘパリンが最も好ましい。Antithrombotic substances include anticoagulants such as hirudin and heparin, plasminogen activators such as t-PA and urokinase, fibrinolytic enzymes such as plasmin and subtilisin, and antiplatelet agents such as prostacyclin and aspirin. It may be used, but heparin is most preferred.
【００２３】以上のような処理によって、延伸ＰＴＦＥ
チューブからなる多孔質チューブの孔内部の壁面を含む
全ての表面に生体組織誘導性物質を共有結合により固定
し、要すれば少なくとも血液と接するチューブの表面に
抗血栓性物質を固定した、移植後の開存性が改善された
人工血管が得られる。By the above treatment, expanded PTFE
A living tissue-inducing substance is covalently immobilized on all surfaces including the inner wall surface of the porous tube made of a tube, and if necessary, an antithrombotic substance is immobilized on the surface of the tube that is in contact with at least blood. An artificial blood vessel with improved patency of
【００２４】[0024]
【実施例】実施例１ 以下の実施・比較例でのべるｅＰＴＦＥの繊維長とは、
走査型電子顕微鏡で測定した結節間距離の平均値であ
る。内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥ
ＰＴＦＥチューブ（チューブ１）と内径２．５ｍｍ、外
径３ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チュ
ーブ２）を用意した。各チューブを、−１０℃でアルゴ
ン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト重合チューブ
を得た。重量変化から計算したところ、アクリル酸グラ
フト量は、チューブ１ｃｍあたり、チューブ１では約８
０μｇであり、チューブ２では約１２０μｇあった。ExamplesExample 1 What is the fiber length of ePTFE in the followingexamples for execution and comparison?
It is the average value of the internodal distance measured by a scanning electron microscope. E with inner diameter 2 mm, outer diameter 2.5 mm, fiber length 60 μm
A PTFE tube (tube 1) and an EPTFE tube (tube 2) having an inner diameter of 2.5 mm, an outer diameter of 3 mm and a fiber length of 30 μm were prepared. Each tube was placed in an atmosphere of argon at −10 ° C. and an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After this, the excess acrylic acid and the acrylic acid polymerized were washed and removed to obtain an acrylic acid graft polymerization tube. When calculated from the change in weight, the amount of acrylic acid grafted was about 8 for tube 1 per 1 cm of tube.
It was 0 μg, and in tube 2, it was about 120 μg.
【００２５】各チューブを、Ｎーヒドロキシこはく酸イ
ミド２．１ｇ 、ジシクロヘキシルカルボジイミド１．
９ｇの１,４−ジオキサン溶液１００ｍｌに、０℃で１
２時間浸漬してＮーヒドロキシこはく酸イミドエステル
とした。このエステル化チューブを、フィブロネクチン
３０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬
してフィブロネクチンを共有結合により固定した。重量
変化から計算したところ、チューブ１ｃｍ当たりに結合
しているフィブロネクチンは、チューブ１では約４５μ
ｇで、チューブ２では約７０μｇであった。Each tube was charged with 2.1 g of N-hydroxysuccinimide, dicyclohexylcarbodiimide 1.
To 100 ml of 9 g of 1,4-dioxane solution,
It was immersed for 2 hours to obtain N-hydroxysuccinimide ester. This esterified tube was immersed in 100 ml of a phosphate buffer solution (pH 11) containing 30 mg of fibronectin to fix fibronectin by covalent bond. When calculated from the change in weight, the amount of fibronectin bound per 1 cm of tube was about 45 μm in tube 1.
In tube 2, it was about 70 μg.
【００２６】次に、チューブをトランスフェリン３０ｍ
ｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬してト
ランスフェリンを共有結合により固定した。重量変化か
ら計算したところ、チューブ１ｃｍ当たりに結合してい
るトランスフェリンは、チューブ１では約３５μｇで、
チューブ２では約６０μｇであった。Then, the tube is transferred to 30 m of transferrin.
The transferrin was covalently fixed by immersing it in 100 ml of phosphate buffer (pH 11). When calculated from the change in weight, transferrin bound to 1 cm of tube was about 35 μg in tube 1,
In tube 2, it was about 60 μg.
【００２７】次に、チューブ１を０．０５％ヘパリン
（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結乾
燥し、ヘパリン複合化延伸ＰＴＦＥを得た。重量変化か
ら計算したところ、固定されたヘパリンは、チューブ１
ｃｍあたり約２００μｇであった。最後に、直径２ｍｍ
のステンレス鋼製の棒にチューブ１を挿入し、その上に
チューブ２が完全に密着するように積層して、人工血管
を製造した。Next, the tube 1 was immersed in a 0.05% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE. Fixed heparin was calculated from tube 1 as calculated from the weight change.
It was about 200 μg per cm. Finally, the diameter is 2mm
The tube 1 was inserted into the stainless steel rod of, and the tube 2 was laminated thereon so that the tube 2 was completely adhered to each other to manufacture an artificial blood vessel.
【００２８】実施例２ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。各チューブを、−１０℃でアルゴン雰
囲気下、メチルリチウムのエーテル溶液（１．４Ｍ）２
０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合溶液に
３０分間浸漬した後、溶液だけを除去し、アクリル酸１
ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間反応さ
せた。この後、余剰のアクリル酸と重合したアクリル酸
を洗浄除去し、アクリル酸グラフト体を得た。重量変化
から計算したところ、アクリル酸のグラフト量は、チュ
ーブ１ｃｍあたり、チューブ１では約８０μｇで、チュ
ーブ２では約１２０μｇあった。Example 2 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. Each tube was placed in an atmosphere of argon at −10 ° C. and an ether solution of methyllithium (1.4 M) 2
After soaking in a mixed solution of 0 ml and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution is removed and acrylic acid 1
A solution of g in 20 ml of water was added, and the mixture was reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the grafted amount of acrylic acid was about 80 μg for tube 1 and about 120 μg for tube 2 per cm of tube.
【００２９】各チューブを、Ｎーヒドロキシこはく酸イ
ミド２．１ｇ、ジシクロヘキシルカルボジイミド１．９
ｇの１,４−ジオキサン溶液１００ｍｌに、０℃で１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブを、フィブロネクチン３
０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬し
てフィブロネクチンを共有結合により固定した。重量変
化から計算したところ、チューブ１ｃｍ当たりに結合し
ているフィブロネクチンは、チューブ１では約４５μｇ
で、チューブ２では約７０μｇであった。Each tube was charged with 2.1 g of N-hydroxysuccinimide and 1.9 of dicyclohexylcarbodiimide.
to 100 ml of a solution of 1,4-dioxane in 12 g at 0 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Fibronectin 3 was added to this esterification tube.
Fibronectin was covalently immobilized by immersing in 100 ml of 0 mg phosphate buffer (pH 11). When calculated from the change in weight, the amount of fibronectin bound per 1 cm of tube was approximately 45 μg in tube 1.
In tube 2, the amount was about 70 μg.
【００３０】次に、チューブ１を０．０５％ヘパリン
（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結乾
燥し、ヘパリン複合化延伸ＰＴＦＥを得た。重量変化か
ら計算したところ、固定されたヘパリンは、チューブ１
ｃｍあたり約２００μｇであった。最後に、直径２ｍｍ
のステンレス鋼製の棒にチューブ１を挿入し、その上に
チューブ２が完全に密着するように積層して、人工血管
を製造した。Next, the tube 1 was immersed in a 0.05% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE. Fixed heparin was calculated from tube 1 as calculated from the weight change.
It was about 200 μg per cm. Finally, the diameter is 2mm
The tube 1 was inserted into the stainless steel rod of, and the tube 2 was laminated thereon so that the tube 2 was completely adhered to each other to manufacture an artificial blood vessel.
【００３１】実施例３ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱して両チューブ
を接着させた。この積層チューブを、−１０℃でアルゴ
ン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約１９０μｇあった。Example 3 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, and the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to bond both tubes. This laminated tube was placed under an argon atmosphere at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 190 μg per cm of the tube.
【００３２】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ、ジシクロヘキシルカルボジイミ
ド１．９ｇの１，４−ジオキサン溶液１００ｍｌに１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブを、フィブロネクチン３
０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬し
てフィブロネクチンを共有結合により固定した。重量変
化から計算したところ、結合しているフィブロネクチン
は、チューブ１ｃｍ当たり約１１０μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 0 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Fibronectin 3 was added to this esterification tube.
Fibronectin was covalently immobilized by immersing in 100 ml of 0 mg phosphate buffer (pH 11). Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００３３】実施例４ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱して両チューブ
を接着させた。この積層チューブを、−１０℃でアルゴ
ン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約１９０μｇあった。Example 4 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, and the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to bond both tubes. This laminated tube was placed under an argon atmosphere at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 190 μg per cm of the tube.
【００３４】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ、ジシクロヘキシルカルボジイミ
ド１．９ｇの１、４−ジオキサン溶液１００ｍｌに１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブを、トランスフェリン３
０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬し
てトランスフェリンを共有結合により固定した。重量変
化から計算したところ、結合しているトランスフェリン
は、チューブ１ｃｍ当たり約９３μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 0 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Transfer this esterification tube to transferrin 3
The transferrin was covalently immobilized by immersion in 100 ml of 0 mg phosphate buffer (pH 11). Transferrin bound was approximately 93 μg per cm tube, calculated from weight changes.
【００３５】実施例５ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱して両チューブ
を接着させた。Example 5 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, and the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to bond both tubes.
【００３６】このチューブを、−１０℃でアルゴン雰囲
気下、メチルリチウムのエーテル溶液（１．４Ｍ）２０
ｍｌとヘキサメチルリン酸アミド２ｍｌの混合溶液に３
０分間浸漬した後、溶液だけを除去し、アクリル酸１ｇ
の水２０ｍｌ中溶液を加え、８０℃で１０時間反応させ
た。この後、余剰のアクリル酸と重合したアクリル酸を
洗浄除去し、アクリル酸グラフト体を得た。重量変化か
ら計算したところ、アクリル酸のグラフト量は、チュー
ブ１ｃｍあたり約１９０μｇあった。This tube was placed in an ether atmosphere (1.4M) of methyllithium 20 at -10 ° C under an argon atmosphere.
ml and a mixed solution of hexamethylphosphoric acid amide 2 ml
After soaking for 0 minutes, remove only the solution and add 1 g of acrylic acid.
Was added to 20 ml of water, and the mixture was reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 190 μg per cm of the tube.
【００３７】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ、ジシクロヘキシルカルボジイミ
ド１．９ｇの１,４−ジオキサン溶液１００ｍｌに１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブをフィブロネクチン３０
ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬して
フィブロネクチンを共有結合により固定した。重量変化
から計算したところ、結合しているフィブロネクチン
は、チューブ１ｃｍ当たり約１１０μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 0 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Fibronectin 30 was added to this esterified tube.
Fibronectin was covalently immobilized by immersing in 100 ml of phosphate buffer (pH 11). Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００３８】次に、チューブをトランスフェリン３０ｍ
ｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬してト
ランスフェリンを共有結合により固定した。重量変化か
ら計算したところ、チューブに結合しているトランスフ
ェリンは、チューブ１ｃｍ当たり約９３μｇであった。Next, the tube was transferred to 30 m of transferrin.
The transferrin was covalently fixed by immersing it in 100 ml of phosphate buffer (pH 11). Transferrin bound to the tube was calculated to be about 93 μg per cm of the tube as calculated from the change in weight.
【００３９】実施例６ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱して両チューブ
を接着させた。Example 6 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, and the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to bond both tubes.
【００４０】このチューブを、−１０℃でアルゴン雰囲
気下、メチルリチウムのエーテル溶液（１．４Ｍ）２０
ｍｌとヘキサメチルリン酸アミド２ｍｌの混合溶液に３
０分間浸漬した後、溶液だけを除去し、アクリル酸１ｇ
の水２０ｍｌ中溶液を加え、８０℃で１０時間反応させ
た。この後、余剰のアクリル酸と重合したアクリル酸と
パラフィンを洗浄除去し、アクリル酸グラフト体を得
た。重量変化から計算したところ、チューブ１のアクリ
ル酸のグラフト量は、チューブ１ｃｍあたり約１９０μ
ｇあった。This tube was placed in an ether atmosphere of methyllithium (1.4M) 20 at −10 ° C. under an argon atmosphere.
ml and a mixed solution of hexamethylphosphoric acid amide 2 ml
After soaking for 0 minutes, remove only the solution and add 1 g of acrylic acid.
Was added to 20 ml of water, and the mixture was reacted at 80 ° C. for 10 hours. After that, excess acrylic acid polymerized with acrylic acid and paraffin were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted on tube 1 was about 190 μm per 1 cm tube.
There was g.
【００４１】０．０５％フィブロネクチン（牛血漿由
来、日本ハム）、及び０．５％１−エチル−３−（３−
ジメチルアミノプロピル）カルボジイミドの水溶液を１
Ｎ塩酸でｐＨ５に調整し、この溶液にアクリル酸グラフ
ト化延伸ＰＴＦＥチューブを２４時間浸漬した後、水で
洗浄し、フィブロネクチン結合延伸ＰＴＦＥチューブを
得た。重量変化から計算したところ、結合しているフィ
ブロネクチンは、チューブ１ｃｍ当たり約１１０μｇで
あった。0.05% fibronectin (derived from bovine plasma, Nippon Ham), and 0.5% 1-ethyl-3- (3-
1 solution of dimethylaminopropyl) carbodiimide
The pH was adjusted to 5 with N hydrochloric acid, and the acrylic acid-grafted expanded PTFE tube was immersed in this solution for 24 hours and then washed with water to obtain a fibronectin-bonded expanded PTFE tube. Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００４２】最後に、このチューブを０．０２％ヘパリ
ン（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結
乾燥し、ヘパリン複合化延伸ＰＴＦＥを得た。重量変化
から計算したところ、固定されたヘパリンは、チューブ
１ｃｍあたり約２００μｇであった。Finally, this tube was immersed in a 0.02% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE. When calculated from the change in weight, the amount of immobilized heparin was about 200 μg per cm of the tube.
【００４３】実施例７ 内径２ｍｍ、外径３ｍｍ、繊維長６０μｍのＥＰＴＦＥ
チューブを用意した。このチューブを、−１０℃でアル
ゴン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約１２０μｇあった。Example 7 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 60 μm
A tube was prepared. This tube was placed under an argon atmosphere at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 120 μg per cm of the tube.
【００４４】０．０５％フィブロネクチン（牛血漿由
来、日本ハム）、及び０．５％１−エチル−３−（３−
ジメチルアミノプロピル）カルボジイミドの水溶液を１
Ｎ塩酸でｐＨ５に調整し、この溶液にアクリル酸グラフ
ト化延伸ＰＴＦＥを２４時間浸漬した後、水で洗浄し、
フィブロネクチン結合延伸ＰＴＦＥチューブを得た。重
量変化から計算したところ、結合しているフィブロネク
チンは、チューブ１ｃｍ当たり約９０μｇであった。0.05% fibronectin (derived from bovine plasma, Nippon Ham) and 0.5% 1-ethyl-3- (3-
1 solution of dimethylaminopropyl) carbodiimide
The pH was adjusted to 5 with N hydrochloric acid, and the acrylic acid-grafted expanded PTFE was immersed in this solution for 24 hours and then washed with water,
Fibronectin-bound expanded PTFE tubes were obtained. The calculated amount of bound fibronectin was about 90 μg per cm of the tube as calculated from the change in weight.
【００４５】実施例８ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長６０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。各チューブを、−１０℃でアルゴン雰
囲気下、メチルリチウムのエーテル溶液（１．４Ｍ）２
０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合溶液に
３０分間浸漬した後、溶液だけを除去し、アクリル酸１
ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間反応さ
せた。この後、余剰のアクリル酸と重合したアクリル酸
を洗浄除去し、アクリル酸グラフト体を得た。重量変化
から計算したところ、アクリル酸のグラフト量は、チュ
ーブ１ｃｍあたり、チューブ１では約８０μｇで、チュ
ーブ２では約９０μｇあった。Example 8 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a diameter of 60 mm and a fiber length of 60 μm was prepared. Each tube was placed in an atmosphere of argon at −10 ° C. and an ether solution of methyllithium (1.4 M) 2
After soaking in a mixed solution of 0 ml and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution is removed and acrylic acid 1
A solution of g in 20 ml of water was added, and the mixture was reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 80 μg in Tube 1 and about 90 μg in Tube 2 per 1 cm of Tube.
【００４６】０．０５％フィブロネクチン（牛血漿由
来、日本ハム）、及び０．５％１−エチル−３−（３−
ジメチルアミノプロピル）カルボジイミドの水溶液を１
Ｎ塩酸でｐＨ５に調整し、この溶液にアクリル酸グラフ
ト化延伸ＰＴＦＥチューブを２４時間浸漬した後、水で
洗浄し、フィブロネクチン結合延伸ＰＴＦＥチューブを
得た。重量変化から計算したところ、チューブ１ｃｍ当
たりに結合しているフィブロネクチンは、チューブ１で
は約４５μｇで、チューブ２では約５０μｇであった。0.05% fibronectin (derived from bovine plasma, Nippon Ham) and 0.5% 1-ethyl-3- (3-
1 solution of dimethylaminopropyl) carbodiimide
The pH was adjusted to 5 with N hydrochloric acid, and the acrylic acid-grafted expanded PTFE tube was immersed in this solution for 24 hours and then washed with water to obtain a fibronectin-bonded expanded PTFE tube. Fibronectin bound per cm of tube was about 45 μg in tube 1 and about 50 μg in tube 2 as calculated from the change in weight.
【００４７】次に、チューブ１を０．０５％ヘパリン
（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結乾
燥し、ヘパリン複合化延伸ＰＴＦＥチューブを得た。重
量変化から計算したところ、固定されたヘパリンは、チ
ューブ１ｃｍあたり約２００μｇであった。最後に、直
径２ｍｍのステンレス鋼製の棒にチューブ１を挿入し、
その上にチューブ２が完全に密着するように積層して、
人工血管を製造した。Next, the tube 1 was immersed in a 0.05% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE tube. When calculated from the change in weight, the amount of immobilized heparin was about 200 μg per cm of the tube. Finally, insert the tube 1 into a 2 mm diameter stainless steel rod,
Stack the tube 2 on top of it,
An artificial blood vessel was manufactured.
【００４８】実施例９ 内径２ｍｍ、外径２．５ｍｍ、繊維長３０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。各チューブを、−１０℃でアルゴン雰
囲気下、メチルリチウムのエーテル溶液（１．４Ｍ）２
０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合溶液に
３０分間浸漬した後、溶液だけを除去し、アクリル酸１
ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間反応さ
せた。この後、余剰のアクリル酸と重合したアクリル酸
を洗浄除去し、アクリル酸グラフト体を得た。重量変化
から計算したところ、アクリル酸のグラフト量は、チュ
ーブ１ｃｍ当たり、チューブ１では約１００μｇで、チ
ューブ２では約１２０μｇあった。Example 9 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 30 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. Each tube was placed in an atmosphere of argon at −10 ° C. and an ether solution of methyllithium (1.4 M) 2
After soaking in a mixed solution of 0 ml and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution is removed and acrylic acid 1
A solution of g in 20 ml of water was added, and the mixture was reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of grafted acrylic acid was about 100 μg for tube 1 and about 120 μg for tube 2 per cm of tube.
【００４９】０．０５％フィブロネクチン（牛血漿由
来、日本ハム）、及び０．５％１−エチル−３−（３−
ジメチルアミノプロピル）カルボジイミドの水溶液を１
Ｎ塩酸でｐＨ５に調整し、この溶液にアクリル酸グラフ
ト化延伸ＰＴＦＥチューブを２４時間浸漬した後、水で
洗浄し、フィブロネクチン結合延伸ＰＴＦＥチューブを
得た。重量変化から計算したところ、チューブ１ｃｍ当
たりに結合しているフィブロネクチンは、チューブ１で
は約５０μｇで、チューブ２では約７０μｇであった。0.05% fibronectin (derived from bovine plasma, Nippon Ham), and 0.5% 1-ethyl-3- (3-
1 solution of dimethylaminopropyl) carbodiimide
The pH was adjusted to 5 with N hydrochloric acid, and the acrylic acid-grafted expanded PTFE tube was immersed in this solution for 24 hours and then washed with water to obtain a fibronectin-bonded expanded PTFE tube. When calculated from the change in weight, the amount of fibronectin bound per 1 cm of tube was about 50 μg in tube 1 and about 70 μg in tube 2.
【００５０】次に、チューブ１を０．０５％ヘパリン
（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結乾
燥し、ヘパリン複合化延伸ＰＴＦＥチューブを得た。重
量変化から計算したところ、固定されたヘパリンは、チ
ューブ１ｃｍあたり約２００μｇであった。最後に、直
径２ｍｍのステンレス鋼製の棒にチューブ１を挿入し、
その上にチューブ２が完全に密着するように積層して、
人工血管を製造した。Next, the tube 1 was immersed in a 0.05% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE tube. When calculated from the change in weight, the amount of immobilized heparin was about 200 μg per cm of the tube. Finally, insert the tube 1 into a 2 mm diameter stainless steel rod,
Stack the tube 2 on top of it,
An artificial blood vessel was manufactured.
【００５１】比較例１ 内径２ｍｍ、外径３ｍｍ、繊維長３０μｍのＥＰＴＦＥ
チューブを用意した。Comparative Example 1 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 30 μm
A tube was prepared.
【００５２】比較例２ 内径２ｍｍ、外径３ｍｍ、繊維長６０μｍのＥＰＴＦＥ
チューブを用意した。Comparative Example 2 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 60 μm
A tube was prepared.
【００５３】比較例３ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱して両チューブ
を接着させた。Comparative Example 3 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, and the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to bond both tubes.
【００５４】比較例４ 内径２ｍｍ、外径３ｍｍ、繊維長３０μｍのＥＰＴＦＥ
チューブを用意した。このチューブを−１０℃でアルゴ
ン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約２１０μｇであった。Comparative Example 4 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 30 μm
A tube was prepared. This tube was placed in an atmosphere of argon at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 210 μg per cm of the tube.
【００５５】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ、ジシクロヘキシルカルボジイミ
ド１．９ｇの１,４−ジオキサン溶液１００ｍｌに１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブをフィブロネクチン３０
ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬して
フィブロネクチンを共有結合により固定した。重量変化
から計算したところ、結合しているフィブロネクチン
は、チューブ１ｃｍ当たり約１１０μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 12 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Fibronectin 30 was added to this esterified tube.
Fibronectin was covalently immobilized by immersing in 100 ml of phosphate buffer (pH 11). Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００５６】比較例５ 内径２ｍｍ、外径３ｍｍ、繊維長３０μｍのＥＰＴＦＥ
チューブを用意した。このチューブを、−１０℃でアル
ゴン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約２１０μｇであった。Comparative Example 5 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 30 μm
A tube was prepared. This tube was placed under an argon atmosphere at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 210 μg per cm of the tube.
【００５７】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ 、ジシクロヘキシルカルボジイ
ミド１．９ｇの１、４−ジオキサン溶液１００ｍｌに１
２時間浸漬してＮーヒドロキシこはく酸イミドエステル
とした。このエステル化チューブを、フィブロネクチン
３０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬
してフィブロネクチンを共有結合により固定した。重量
変化から計算したところ、結合しているフィブロネクチ
ンは、チューブ１ｃｍ当たり約１１０μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 0 ° C.
It was immersed for 2 hours to obtain N-hydroxysuccinimide ester. This esterified tube was immersed in 100 ml of a phosphate buffer solution (pH 11) containing 30 mg of fibronectin to fix fibronectin by covalent bond. Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００５８】最後に、このチューブを０．０２％ヘパリ
ン（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結
乾燥し、ヘパリン複合化延伸ＰＴＦＥを得た。重量変化
から計算したところ、固定されたヘパリンは、チューブ
１ｃｍあたり約２００μｇであった。Finally, this tube was immersed in a 0.02% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE. When calculated from the change in weight, the amount of immobilized heparin was about 200 μg per cm of the tube.
【００５９】比較例６ 内径２ｍｍ、外径３ｍｍ、繊維長６０μｍのＥＰＴＦＥ
チューブを用意した。このチューブを、−１０℃でアル
ゴン雰囲気下、メチルリチウムのエーテル溶液（１．４
Ｍ）２０ｍｌとヘキサメチルリン酸アミド２ｍｌの混合
溶液に３０分間浸漬した後、溶液だけを除去し、アクリ
ル酸１ｇの水２０ｍｌ中溶液を加え、８０℃で１０時間
反応させた。この後、余剰のアクリル酸と重合したアク
リル酸を洗浄除去し、アクリル酸グラフト体を得た。重
量変化から計算したところ、アクリル酸のグラフト量
は、チューブ１ｃｍあたり約１２０μｇであった。Comparative Example 6 EPTFE having an inner diameter of 2 mm, an outer diameter of 3 mm and a fiber length of 60 μm
A tube was prepared. This tube was placed under an argon atmosphere at −10 ° C. to prepare an ether solution of methyllithium (1.4
After dipping in a mixed solution of 20 ml of M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes, only the solution was removed, and a solution of 1 g of acrylic acid in 20 ml of water was added and reacted at 80 ° C. for 10 hours. After that, the excess acrylic acid and the polymerized acrylic acid were washed and removed to obtain an acrylic acid graft body. When calculated from the change in weight, the amount of acrylic acid grafted was about 120 μg per 1 cm of the tube.
【００６０】このチューブを、０℃でＮーヒドロキシこ
はく酸イミド２．１ｇ、ジシクロヘキシルカルボジイミ
ド１．９ｇの１,４−ジオキサン溶液１００ｍｌに１２
時間浸漬してＮーヒドロキシこはく酸イミドエステルと
した。このエステル化チューブを、フィブロネクチン３
０ｍｇのリン酸緩衝液（ｐＨ１１）１００ｍｌに浸漬し
てフィブロネクチンを共有結合により固定した。重量変
化から計算したところ、結合しているフィブロネクチン
は、チューブ１ｃｍ当たり約９０μｇであった。This tube was added to 100 ml of a 1,4-dioxane solution containing 2.1 g of N-hydroxysuccinimide and 1.9 g of dicyclohexylcarbodiimide at 12 ° C.
It was immersed for a time to obtain N-hydroxysuccinimide ester. Fibronectin 3 was added to this esterification tube.
Fibronectin was covalently immobilized by immersing in 100 ml of 0 mg phosphate buffer (pH 11). The calculated amount of bound fibronectin was about 90 μg per cm of the tube as calculated from the change in weight.
【００６１】最後に、このチューブを０．０２％ヘパリ
ン（ＳＡＮＯＦＩ社）水溶液に１０分浸漬した後、凍結
乾燥し、ヘパリン複合化延伸ＰＴＦＥチューブを得た。
重量変化から計算したところ、固定されたヘパリンは、
チューブ１ｃｍあたり約２００μｇであった。Finally, this tube was immersed in a 0.02% heparin (SANOFI) aqueous solution for 10 minutes and then freeze-dried to obtain a heparin-composited expanded PTFE tube.
When calculated from the change in weight, the fixed heparin was
The amount was about 200 μg per 1 cm of the tube.
【００６２】比較例７ 内径２ｍｍ、外径２．５ｍｍ、繊維長６０μｍのＥＰＴ
ＦＥチューブ（チューブ１）と内径２．５ｍｍ、外径３
ｍｍ、繊維長３０μｍのＥＰＴＦＥチューブ（チューブ
２）を用意した。直径２ｍｍのステンレス鋼製の棒にチ
ューブ１を挿入し、その上にチューブ２が完全に密着す
るように積層し、３４０℃で２０分加熱し接着させた。
このチューブを、−１０℃でアルゴン雰囲気下、メチル
リチウムのエーテル溶液（１．４Ｍ）２０ｍｌとヘキサ
メチルリン酸アミド２ｍｌの混合溶液に３０分間浸漬し
た後、溶液だけを除去し、アクリル酸１ｇの水２０ｍｌ
中溶液を加え、８０℃で１０時間反応させた。この後、
余剰のアクリル酸と重合したアクリル酸を洗浄除去し、
アクリル酸グラフト体を得た。重量変化から計算したと
ころ、アクリル酸のグラフト量は、チューブ１ｃｍあた
り約１９０μｇあった。Comparative Example 7 EPT having an inner diameter of 2 mm, an outer diameter of 2.5 mm and a fiber length of 60 μm
FE tube (tube 1) with inner diameter 2.5 mm, outer diameter 3
An EPTFE tube (tube 2) having a length of 30 mm and a fiber length of 30 μm was prepared. The tube 1 was inserted into a stainless steel rod having a diameter of 2 mm, the tube 2 was laminated thereon so that the tube 2 was completely adhered thereto, and heated at 340 ° C. for 20 minutes to be adhered.
This tube was immersed in a mixed solution of 20 ml of a methyllithium ether solution (1.4 M) and 2 ml of hexamethylphosphoric acid amide for 30 minutes at −10 ° C. under an argon atmosphere, and then only the solution was removed to obtain 1 g of acrylic acid. 20 ml of water
The medium solution was added, and the mixture was reacted at 80 ° C. for 10 hours. After this,
Wash away excess acrylic acid and acrylic acid polymerized,
An acrylic acid graft product was obtained. When calculated from the change in weight, the amount of acrylic acid grafted was about 190 μg per cm of the tube.
【００６３】０．０５％フィブロネクチン（牛血漿由
来、日本ハム）、及び０．５％１−エチル−３−（３−
ジメチルアミノプロピル）カルボジイミドの水溶液を１
Ｎ塩酸でｐＨ５に調整し、この溶液にアクリル酸グラフ
ト化延伸ＰＴＦＥチューブを２４時間浸漬した後、水で
洗浄し、フィブロネクチン結合延伸ＰＴＦＥチューブを
得た。重量変化から計算したところ、結合しているフィ
ブロネクチンは、チューブ１ｃｍ当たり約１１０μｇで
あった。0.05% fibronectin (derived from bovine plasma, Nippon Ham), and 0.5% 1-ethyl-3- (3-
1 solution of dimethylaminopropyl) carbodiimide
The pH was adjusted to 5 with N hydrochloric acid, and the acrylic acid-grafted expanded PTFE tube was immersed in this solution for 24 hours and then washed with water to obtain a fibronectin-bonded expanded PTFE tube. Fibronectin bound was about 110 μg per cm of the tube as calculated from the change in weight.
【００６４】次に、０．０５％トランスフェリン、及び
０．５％１−エチル−３−（３−ジメチルアミノプロピ
ル）カルボジイミドの水溶液を１Ｎ塩酸でｐＨ５に調整
し、この溶液にアクリル酸グラフト化延伸ＰＴＦＥチュ
ーブを２４時間浸漬した後、水で洗浄し、トランスフェ
リン結合延伸ＰＴＦＥチューブを得た。重量変化から計
算したところ、結合しているフィブロネクチンは、チュ
ーブ１ｃｍ当たり約９５μｇであった。Next, an aqueous solution of 0.05% transferrin and 0.5% 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide was adjusted to pH 5 with 1N hydrochloric acid, and this solution was acrylic acid-grafted and stretched. The PTFE tube was soaked for 24 hours and then washed with water to obtain a transferrin-bonded expanded PTFE tube. The calculated amount of bound fibronectin was about 95 μg per cm of the tube as calculated from the change in weight.
【００６５】実施例および比較例の人工血管（各３ｃ
ｍ）を、ウサギの頚動脈に置換移植した。結果を下記表
１に示す。血液に接触する面の繊維長が６０μｍでその
他の部分の繊維長が３０μｍで、且つフィブロネクチン
とヘパリンの両方を固定している人工血管の開存率が優
れており、特にヘパリンを内層のみに固定した人工血管
の開存率が高く、１００％であった。また、繊維長３０
μｍの層を含む人工血管以外では、移植後に人工血管が
湾曲したり、周囲組織との癒着が激しい例が多く存在し
た。Artificial blood vessels of Examples and Comparative Examples (3 c each)
m) was replaced and transplanted into the carotid artery of a rabbit. The results are shown in Table 1 below. The fiber length of the surface that comes into contact with blood is 60 μm and the fiber length of other parts is 30 μm, and the patency of the artificial blood vessel that fixes both fibronectin and heparin is excellent, and especially heparin is fixed only to the inner layer. The patency rate of the artificial blood vessel was 100%. Also, fiber length 30
In addition to the artificial blood vessel including the layer of μm, there were many cases where the artificial blood vessel was curved after transplantation or the adhesion with surrounding tissues was severe.
【００６６】また、内皮細胞で被覆されている面積を測
定した結果、フィブロネクチンやトランスフェリンをＮ
ーヒドロキシこはく酸イミド（活性エステル法）を用い
て固定した人工血管において被覆面積が高く、特にフィ
ブロネクチンとトランスフェリンの両方を固定した人工
血管で最も被覆率が高かった。また、繊維長３０μｍの
層を含む人工血管以外では、移植後に人工血管が湾曲し
たり、周囲組織との癒着が激しい例が多く存在した。As a result of measuring the area covered with endothelial cells, fibronectin and transferrin were detected in N
The area of the artificial blood vessel fixed with -hydroxysuccinimide (active ester method) was high, and the area of the artificial blood vessel with both fibronectin and transferrin was highest. In addition to the artificial blood vessel including a layer having a fiber length of 30 μm, there were many cases where the artificial blood vessel was curved after transplantation and the adhesion with surrounding tissues was severe.
【００６７】一方、０．５％１−エチル−３−（３−ジ
メチルアミノプロピル）カルボジイミド（ＷＳＣ法）を
用いて組織誘導性物質を固定した比較例の人工血管で
は、内皮細胞の被覆率は低かった。また、繊維長３０μ
ｍの層を含む人工血管以外では、移植後に人工血管が湾
曲したり、周囲組織との癒着が激しい例が多く存在し
た。On the other hand, in the artificial blood vessel of the comparative example in which the tissue-inducing substance was fixed using 0.5% 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (WSC method), the coverage of endothelial cells was It was low. The fiber length is 30μ
In addition to the artificial blood vessel including the m layer, there were many cases where the artificial blood vessel was curved after transplantation and had strong adhesion with surrounding tissues.
【００６８】[0068]
【表１】[Table 1]
【００６９】[0069]
【発明の効果】従来より延伸ポリテトラフルオロエチレ
ンのチューブが人工血管として用いられている。しかし
ながらこの材料自体は抗血栓性が十分ではなく、特に内
径４ｍｍ以下の小口径領域では血栓による閉塞が頻発す
る。本発明によれば、延伸ＰＴＦＥの多孔質チューブの
孔内部の壁面を含む全ての表面に化学的に官能基を導入
し、その官能基を導入した層に生体組織誘導性物質を共
有結合により固定し、要すれば抗血栓性物質を少なくと
も血液と接触する表面に固定したことにより、移植後の
開存性が改善された人工血管が得られる。EFFECTS OF THE INVENTION A tube of expanded polytetrafluoroethylene has been conventionally used as an artificial blood vessel. However, this material itself does not have sufficient antithrombogenicity, and in particular, in a small diameter region having an inner diameter of 4 mm or less, clogging with thrombus frequently occurs. According to the present invention, a functional group is chemically introduced to all surfaces including the wall surface inside the pores of the expanded PTFE porous tube, and the biological tissue-inducing substance is covalently fixed to the layer having the functional group introduced. However, if necessary, by fixing an antithrombotic substance on at least the surface that comes into contact with blood, an artificial blood vessel with improved patency after transplantation can be obtained.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15386896AJP3014324B2 (en) | 1996-02-15 | 1996-06-14 | Artificial blood vessel |
| CA002197375ACA2197375C (en) | 1996-02-15 | 1997-02-12 | Artificial blood vessel |
| US08/800,685US6053939A (en) | 1996-02-15 | 1997-02-14 | Artificial blood vessel |
| AU12682/97AAU709305B2 (en) | 1996-02-15 | 1997-02-14 | Artificial blood vessel |
| DE69725198TDE69725198T2 (en) | 1996-02-15 | 1997-02-14 | Artificial blood vessel |
| EP97102455AEP0790042B1 (en) | 1996-02-15 | 1997-02-14 | Artificial blood vessel |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2782396 | 1996-02-15 | ||
| JP8-27823 | 1996-02-15 | ||
| JP15386896AJP3014324B2 (en) | 1996-02-15 | 1996-06-14 | Artificial blood vessel |
| Publication Number | Publication Date |
|---|---|
| JPH09276395Atrue JPH09276395A (en) | 1997-10-28 |
| JP3014324B2 JP3014324B2 (en) | 2000-02-28 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15386896AExpired - Fee RelatedJP3014324B2 (en) | 1996-02-15 | 1996-06-14 | Artificial blood vessel |
| Country | Link |
|---|---|
| JP (1) | JP3014324B2 (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013538281A (en)* | 2010-09-21 | 2013-10-10 | ダブリュ．エル．ゴア アンド アソシエイツ，インコーポレイティド | Stretchable functional TFE copolymer fine powder, stretch functional product obtained therefrom, and reaction of the stretch product |
| AU2014355414B2 (en)* | 2013-11-28 | 2017-06-01 | Toray Industries, Inc. | Antithrombotic material |
| WO2019106996A1 (en)* | 2017-11-30 | 2019-06-06 | 株式会社日立製作所 | Immunoisolation device |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013538281A (en)* | 2010-09-21 | 2013-10-10 | ダブリュ．エル．ゴア アンド アソシエイツ，インコーポレイティド | Stretchable functional TFE copolymer fine powder, stretch functional product obtained therefrom, and reaction of the stretch product |
| JP2016041826A (en)* | 2010-09-21 | 2016-03-31 | ダブリュ．エル．ゴア アンド アソシエイツ，インコーポレイティドＷ．Ｌ． Ｇｏｒｅ ＆ Ａｓｓｏｃｉａｔｅｓ， Ｉｎｃｏｒｐｏｒａｔｅｄ | Expandable functional tfe copolymer fine powder, expanded functional products obtained therefrom and reaction of expanded products |
| AU2014355414B2 (en)* | 2013-11-28 | 2017-06-01 | Toray Industries, Inc. | Antithrombotic material |
| WO2019106996A1 (en)* | 2017-11-30 | 2019-06-06 | 株式会社日立製作所 | Immunoisolation device |
| Publication number | Publication date |
|---|---|
| JP3014324B2 (en) | 2000-02-28 |
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| EP0790042B1 (en) | Artificial blood vessel | |
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| Date | Code | Title | Description |
|---|---|---|---|
| R250 | Receipt of annual fees | Free format text:JAPANESE INTERMEDIATE CODE: R250 | |
| LAPS | Cancellation because of no payment of annual fees |