JP2008029866A - 精製粘膜下組織からの管状移植片 - Google Patents
精製粘膜下組織からの管状移植片 Download PDFInfo
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Abstract
【解決手段】本発明の人工器官は所望の長さ、管壁厚さ又は直径で製造される。本発明の方法により製造された構造物は、動脈、静脈、尿管、尿道、シャント又は従順で組織適合性の或る管が必要な全ての用途における移植片として使用できる。移植片人工器官の製造は一般的に、精製されたコラーゲン系のマトリックス構造物の複数のシートをマンドレルの周囲に捲回させ、マンドレル上の組織を圧縮及び乾燥させ、その後、この構造物を、来るべき使用のために、マンドレルから取り外す。
【選択図】 図1
Description
A. 多層重複領域を有する精製粘膜下組織の管を形成するために、マンドレルの周囲全体に精製粘膜下組織のシートを被せるステップと、
B. 重複領域における精製粘膜下組織層を互いに固着させるステップと、
C. 精製粘膜下組織の第2の管の継目が縫合により密閉されている、精製粘膜下組織の第2の管を形成するために、(場合により)精製粘膜下組織の第2のシートを精製粘膜下組織の管に被せるステップと、
D.脱水条件下で、精製粘膜下組織の複数層を圧縮するステップとからなる。
本発明は、形成された管状人工器官の壁が、管の内腔から外表面への直接的な通路を与える穿孔を全く有しないように、精製粘膜下組織のシートから多層管状移植片構造物を構築することを斟酌する。本発明の多層管状人工器官は、管状人工器官の漏れ又は故障無しに脈管用途で使用されるべき、十分な強度と耐久性を有する。
バイオバーデン(Bioburden):所定量の材料上に及び/又は材料中に発見される、生存微生物の個数(コロニー生成単位(CFU)で報告される)を意味する。微生物の代表例は、細菌、真菌及びこれらの胞子などである。
殺菌:材料のバイオバーデンの低減を意味する。
滅菌:材料の所定の区域上及び/又は区域中に1個の生存微生物(CFU)を有する可能性が1億分の1以下であるようなバイオバーデンを材料が有する状態を意味する。
発熱物質:ホスト内に導入された後、熱性反応を生じる物質を意味する。
内毒素:グラム陰性細菌の細胞壁の一部である特別の発熱物質を意味する。内毒素は、細菌及び汚染物質から頻繁に注がれる。
精製:材料から生じる1個以上の汚染物質(例えば、材料から自然に生じる汚染物質及び/又は材料上に発生する微生物及びこれらの成分)を除去するための材料の処理操作を意味する。例えば、汚染物質は、毒性、感染性、発熱性、刺激性、反応性、溶血作用、発癌性及び/又は免疫原生などの原因となることが知られた物質である。
生体適合性:国際標準化機関(ISO)の規格No.10993及び/又は米国薬局方(USP)23及び/又は米国食品医薬品局(FDA)ブルーブックメモランダムNo.G95-1(表題:Use of International Standard ISO-10993, Biological Evaluation of Medical Device Part-1: Evaluation and Testing)に記載されるような生体適合性試験に合格する材料の能力を意味する。一般的に、これらの試験は、材料の毒性、感染性、発熱性、刺激性、反応性、溶血作用、発癌性及び/又は免疫原生に関して検査する。多数の患者に導入したときに、生体適合性構造物又は材料は、有害反応又は有害応答を起こさない。更に、生体適合性は、プリオン、界面活性剤、オリゴヌクロチドなどのようなその他の汚染物質及びその他の生体適合性に悪影響を及ぼす物質又は汚染物質により悪影響を受ける。
汚染物質:材料上の、又は材料に付着した若しくは材料内の望ましからざる物質を意味する。これは例えば、バイオバーデン、内毒素、殺菌剤のような処理剤、血液、血液成分、ウイルス、DNA、RNA、胞子、望ましからざる組織層の断片、細胞残屑及び粘膜などである。しかし、これらに限定されない。
粘膜下組織:動物の消化管、気道、尿路及び生殖器管の大部分の粘膜下に生じるコラーゲン含有結合組織を意味する。
或る実施態様では、構造物の製造方法は、適当な直径の透水性で中空のマンドレルを選定し、そして、マンドレルに臍帯を螺旋状に捲回することからなる。その後、所望枚数の精製粘膜下組織外被を被せる。“精製粘膜下組織外被”は、マンドレルの周囲に約360゜にわたって巻き付けられた精製粘膜下組織片として定義される。一般的に、1枚又は2枚の外被は、下記のようにして製造された後で、約1000〜2000mmHgの破壊強度をもたらす。
或る実施態様では、最後のシートが緩まないようにするために、捲回マトリックス構造物の継目を“スポット溶接”することができる。或る実施態様によれば、継目はグルタルアルデヒドのような架橋剤でスポット溶接される。X%のグルタルアルデヒド(又はその他の架橋剤若しくは接着剤)で濡らされたQチップを、継目を形成する重複領域に沿って擦り付ける。Xの値は約0.1〜1.0%、好ましくは約0.5%である。しかし、継目幅、グルタルアルデヒド濃度及び破裂圧力を決定するターン回数との間には関係がある。
スポット溶接された精製粘膜下組織ストリップに対するホストの応答を評価するために、離乳子畜のラットモデルを使用した。麻酔薬を導入し、顔面マスクを介してメタファン(metafane)を投与することにより麻酔状態を維持した。腹の腹面を刈り込み、無菌手術のための剃毛面を生成した。各腹の4分円に縦長の皮膚切開部を形成した。次いで、直截的な解剖により各ラットの皮下組織に、左右相称の皮下ポケットを形成した。面積1cm2の試験片を皮下的に各ポケット内に配置し、1個の5−0ポリプロピレン縫合により下部の筋膜の所定位置に固定させた。皮膚切開部を5−0ポリプロピレンによる単純な中断縫合により閉鎖した。この実験には、24匹のラットを使用した。
形態学的評価のために摘出されたサンプルは24時間かけてトランプ(Trump)の固定液中で固定させ、その後、リン酸塩緩衝液内に配置した。光学顕微鏡用の試験片をパラフィン内に埋め込み、2〜3μmの薄片に切り分けた。全体的な形態評価のために、薄片をヘマトキシリン・エオシン(H&E)染色法で染色し、石灰化評価のためにフォン・コッサ(Von Kossa)染色法により染色した。
この実験の目的は、外径5.0mmの移植片を形成することである。多数の孔を有する、直径4mmの中空マンドレルに、重複しないように臍帯を螺旋状に捲回した。その後、精製粘膜下組織を2回半捲回し、精製粘膜下組織の管を生成し、重複部分を、一方のグループでは、グルタルアルデヒドで固着させ、別のグループでは、縫合により固着させた。次いで、精製粘膜下組織の第2のシートを、精製粘膜下組織の管の周囲に被せ、精製粘膜下組織の第2のシートの両縁を一緒に縫合した。捲回終了後、マンドレルの一方の端部を真空ポンプに接続し、マンドレルの他方の端部を閉鎖した。マンドレル内に生じた真空状態は、精製粘膜下組織の複数の層を一緒に堅固に圧縮し、精製粘膜下組織から水分を吸い出す。この処理には約24時間必要である。
破裂試験の前に、各精製粘膜下組織移植片を37℃の0.9%生理食塩水に24時間浸漬させた。この処理の目的は継目の耐久性を決定することである。
破裂強度を決定するために、管状精製粘膜下組織移植片の一方の端部を、空気圧を加えるために使用される取付部品に取付けた。管状精製粘膜下組織移植片の他方の端部を縫合により閉鎖し、空気圧を加圧した。圧力対時間を連続的に記録することにより、破裂圧力を正確に識別することができる。本発明の目標は、1000mmHg以上の破裂圧力である。好結果をもたらす製造技術は、離層を起こすこと無く、(任意の圧力で)破裂を生じるような技術である。この静的試験に合格した管状精製粘膜下組織移植片は引き続き、拍動試験に付される。
管状精製粘膜下組織移植片の危険期間は、再建の初期段階にある場合及び所定の静圧(拡張期圧)及び拍動圧を伴う温血に暴露される場合、移植後の最初の数週間である。この期間中、移植片が、再建の初期段階において、その強度を保持しているか否か知ることが最も重要である。従って、37℃の生理食塩水中で、移植片構造物について拍動圧試験を行う。1/秒の周期の、200/150mmHgの拍動圧を使用する。移植片が静圧破裂強度について試験された後、2週間にわたって、この拍動試験を続ける。
穿孔マンドレルに、精製粘膜下組織を2回半捲回させ、そして、この構造物を8時間真空乾燥させることにより、多層積層構造の構造物を形成した。この構造物をマンドレルから取り出し、20分間にわたって再水和させ、そして、イヌに尿管の人工器官として外科的に移植した。この人工器官は漏出を起こすこと無く尿を導通し、移植された箇所で癒着し、新たな尿管を生成した。
Claims (1)
- 第1の縁部と第2の反対縁部を有し、管の形状に成形された、粘膜下組織源から採取されたコラーゲン系のマトリックス構造物のシートからなり、
複数の層が重複した領域を画成するために、前記第1のシートの前記第2の反対縁部は前記第1のシートの前記第1の縁部を超えて延びており、
前記重複領域の複数の層は相互に固着されている、
ことを特徴とする単一的な多層移植片人工器官。
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JP52599298A Expired - Fee Related JP4084420B2 (ja) | 1996-12-10 | 1997-12-10 | 管状粘膜下組織移植片構成物 |
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KR (2) | KR100646121B1 (ja) |
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