Name: autologous microfractured and purified adipose tissue for arthroscopic management of osteochondral lesions of the talus
Uploaded: 2018-01-23T13:00:00.000+00:00
Duration: 7 min 24 s
Description: Watch this Scientific Journal Video about Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus at JoVE.com

手順は、Lipogems システムを使用して実行します。

すべてのプロシージャ実行 1964 ヘルシンキ宣言とその後の補正または匹敵する倫理と制度や国の研究会では、倫理的な基準を含む人間被験者研究基準。
1. 医療の歴史
<ol><li>患者の詳細な履歴と臨床検査を開始します。 注: OLT 必要があります常に疑われる足首の不安定性の場合、剛性、痛み、関節の締まりと膨潤に関連付けられている繰り返しの捻挫。また、多くの場合、OLT は炎症や血管疾患、神経障害や神経疾患、糖尿病などの全身性疾患の既往が関連づけられます。薬や治療に影響を与える可能性があります医療の問題の使用を評価され、考慮する必要があります。</li>
</ol>2. 臨床検査
<ol><li>足首や後肢足の変形を強調する起立位置に患者を評価します。筋肉や腱関数と足首の運動 (ROM) の範囲を評価します。最大屈伸時に特にびまん性の圧痛が存在する多くの場合、関節の関節のレベルでタッチ敏感なエリアに遭遇することも珍しくありません。</li>
	<li>併用の足関節外側不安定性を識別するために前方および後方引き出しテストを実行します。</li>
	<li>術前相談の間に次の臨床的・機能的スコアを記録: アメリカの整形外科の足と足首の社会 (AOFAS) 足首と後肢足スコア15、視覚アナログ スケール (VAS) 痛みのスコア16と 12 項目の短い形式健康調査 (SF-12)17。</li>
</ol>3. 放射線評価
<ol><li>両側荷重足と足首のレントゲン写真を実行します。これは、従来の重量軸受け前後 (AP) ほぞと足首の横方向荷重ビューので構成されます。距骨のより良い視覚化のための内部の回転の 15 度と中立的な立場では、AP のほぞを実行します。 注: OLT の 50% だけはレントゲン写真; と診断されることができます。大きな病変の場合、見性に囲まれた戸建骨の領域は指摘18をすることができます。</li>
	<li>従来計算された断層レントゲン写真撮影スキャン (ct) 足首を実行します。CT スキャンにより正確な位置と、病変の大きさも場合に剥離骨片を識別します。CT の弱点は、軟骨の状態を表示する機能です。前の研究は、CT19,20Olt を検出するため, 感度, 特異性 0.81 と 0.99 を示した。</li>
	<li>足首の磁気共鳴画像 (MRI) を実行します。MRI は、軟骨と軟骨下骨評価の基本です。また、MRI は電離放射線を必要としない、軟部組織のより良い視覚化をことができます。文献は、感度と Olt21,22の検出に 96% の特異性を報告します。</li>
</ol>4. 手術手技
<ol><li>収穫および脂肪組織の加工
	<ol>
<li>クラインのソリューションの準備: 10 mg/ml トリアムシノロン 0.1 mL、10 mL の 8.4% 重炭酸ナトリウム、エピネフリン 1: 1000 の 1 mL 50 mL の 1% リドカインの 9 グラム/L の生理食塩水 1 L。</li>
		<li>メスの刃で約 0.5 cm の 2 つパラ臍切開を作成します。クライン溶液約 300 mL を 60 mL 注射器を用いた 18 G 針 (図 1) 切開を通じて腹部の皮下脂肪組織に注入します。</li>
		<li>20 mL の注射器に接続されている、および処理キット (図 2) に導入 13 G 鈍カニューレを使用して ADSCs の 40-45 mL を収穫します。通常、ペリ臍周辺地域で収穫を行います。</li>
		<li>クローズド システムで 100-130 mL lipoaspirate を挿入します。最初のクラスター削減を取得する大きなフィルターを介してデバイスに、lipoaspirate をプッシュします。同時に対応する食塩量は無駄バッグに向かって終了します。重要な役割は、オイル、血、生理食塩水の一時的なエマルジョンを取得するステンレス大理石による演奏です。食塩水の重力の対向流によって油の残留物や汚染された血液を削除します。</li>
		<li>この洗浄のステップ (クリアと lipoaspirate 黄色液流動が表示されます) 後、生理食塩水のフラックスを停止し、反転装置 (アップ グレー キャップ)、2 つ目の脂肪クラスター削減に 。10 mL 注射器で液を下から押して 2 番目の切削六角形フィルターを介してフローティング脂肪クラスターを押すことにより削減を取得します。デバイスの上部の開口部に接続されている 10 mL 注射器に最終製品を収集します。 注意: 脂肪質のティッシュの処理キット標準 lipofilling テクニックが向上: 実際には、システムから成る閉じた、ブエノス ・ アイレス、低圧円筒システム ペリサイトの偉大な番号を含む流体と均一な製品を得るために。この手順は、軽度の機械的力を介して排他的脂肪細胞の処理と間質血管ニッチの整合性を維持できます。プロセスは、少なくとも心的外傷の可能です、酵素治療または展開しない短時間 (15-20 分) で利用可能な最終製品になります。破損していない vasculostromal のニッチは、治癒過程を助けます。</li>
		<li>ADSCs が収穫された後は、腹部に圧迫包帯を適用します。</li>
	</ol></li>
	<li>手術と脂肪注入
	<ol>
<li>止血帯、大腿部出血を減少しより良い鏡可視化1をレベルの 250 mmHg の圧力で脊髄くも膜下麻酔の下で仰臥位で患者を置きます。</li>
		<li>皮膚の解剖学的ランドマークをそのペンでマークします。ランドマークは、医原性損傷を避けるために不可欠です。 次のマーク (図 3)。 両方の果 (横 (LM) と内側 (MM)) 識別と背筋- と - の底屈足関節前方共同ライン (JL) 前脛骨筋腱 (TAT) とアキレス腱 大伏在静脈、内果前だけを実行します。 浅腓骨神経 (SPN)</li>
		<li>最初に、前脛骨筋腱、ソフト スポットと一致するちょうど内側の前内側ポータルを実行します。このポータルは、アプローチのポータルを表します。ほとんどの場合、足首の背屈でうつ病、表示され明白です。<ol>
<li>ブレードと皮膚のみをカットし、鈍的切離でカプセルを穿孔します。伏在神経と大伏在静脈を避けるために注意してください。大伏在静脈は、神経は約 7.4 mm 横ポータルにポータルに横方向に 9 mm の位置です。これは 2 つの第一次観覧のポータルの 1 つを構成します。</li>
		</ol>
</li>
		<li>共同線をチェック、前外側のポータルは、外側のくるぶしの内側を置き、外側指伸筋腱。 注: 前外側ポータルを実行すると、中間の背側皮膚神経 (浅腓骨神経の側枝); 怪我を防ぐこのため、切断した後、皮膚、必ず鈍的切離。</li>
		<li>サイズと位置を評価するために軟骨を検査します。条件と、palpator と軟骨の品質を評価します。距骨骨軟骨病変は通常ある posteromedially または前外側。<ol>
<li>いくつかの外科医は大きい 4 mm の関節鏡を使用し、計測器をおいて関節の前方の陥凹が 30 ° の角度で広角の 2.7 mm の世界を使用して関節鏡治療を実行します。Talar ドームのほとんどにアクセスする非侵襲的な共同の気晴らしの技術とハイパー足関節底屈を使用できます。</li>
			<li>後部の病変の場合, 経皮的関節をそらすし病変の露出を許可する Hintermann スプレッダーを配置します。Hintermann 拡散機は開放レバー腕以前に配置 talar 骨と脛骨内側または横に、病変側によると 2 K 線に適用されます。横方向の病変の場合脛骨の骨、関節のよりよい気晴らしを達成するために、腓骨を避けることで近位 K ワイヤーを挿入する注意してください。</li>
		</ol>
</li>
		<li>キューレットで病変を準備、破損して不安定な軟骨を取り外し、石灰化層と正規形を作成するために壊死、硬化の骨肩線で病変に含まれています。この手順では、標準的な鏡視下掻爬 (図 4) を使用します。</li>
		<li>Microperforations を実行する丸く外側から病変の内側に軟骨下骨から骨髄幹細胞を刺激します。<ol>
<li>それらの間に約 3 mm の間隔で穿孔を行います。微小欠陥 (図 5) の下に健康な軟骨下骨の軟骨 Pick を使用を誘発します。 注: フィブリン血栓の形成で起因する出血。血液凝固の炎症カスケードの活性化を介して、製品リリース血管作動性メディエーター、成長因子とサイトカイン。これらの要因は、脈管侵襲と間葉系幹細胞の軟骨病変部への移行を促進する力を持っています。線維芽細胞、軟骨細胞、骨芽細胞に分化し、病変の修復の刺激で重要な役割を果たすこれら多能性細胞を刺激します。関節の環境でパラクリン増殖因子は、細胞外マトリックスの形成と線維軟骨の生産を促進します。同じ時に、骨病変部の細胞は徐々 に成熟した骨に置換される幼若骨組織を生成します。</li>
		</ol>
</li>
		<li>吸引と関節が完全に乾くまで綿スポンジを使用して残りの液体のシェーバーを使用して関節内の水を削除します。</li>
	</ol></li>
	<li>脂肪由来幹細胞の注入
	<ol>
<li>脂肪由来幹細胞は、以前 2 つのポータル (前内側や前外側) のいずれかを使用して関節に 4.1.3 の手順で準備の 5-7 mL を注入します。</li>
		<li>ターニケットをリリースします。</li>
	</ol></li>
</ol>5. 術後のケア
<ol><li>完全放電の作動肢での 15 日間の足首の動きは避けてください。</li>
	<li>術後後 15 日目に足首のアクティブおよびパッシブの動きが許可するは、完全に回復し、可動域に到達するまで。</li>
	<li>手術後 30 日間の充電を許可します。</li>
</ol>6. 臨床およびレントゲン写真のフォロー アップ
<ol><li>臨床的に行えた 6 と 12 ヶ月で、その後、毎年患者を評価します。フォロー アップのプロトコルの各終点15,16,17SF 12 VAS 疼痛スコアと後肢足スコア AOFAS 足首から成ります。放射線透過試験には、MRI と運営の足首の CT スキャンが含まれています。</li>
</ol>

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G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteochondral+lesions+of+the+talus:+a+new+magnetic+resonance+grading+system+with+arthroscopic+correlation.">Osteochondral lesions of the talus: a new magnetic resonance grading system with arthroscopic correlation.</a> Arthroscopy. 19 (4), 353-359 (2003).</li><li>Leumann, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+imaging+method+for+osteochondral+lesions+of+the+talus--comparison+of+SPECT-CT+with+MRI.">A novel imaging method for osteochondral lesions of the talus--comparison of SPECT-CT with MRI.</a> Am. J. Sports Med. 39 (5), 1095-1101 (2011).</li><li>Kim, Y. S., Park, E. H., Kim, Y. C., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+outcomes+of+mesenchymal+stem+cell+injection+with+arthroscopic+treatment+in+older+patients+with+osteochondral+lesions+of+the+talus.">Clinical outcomes of mesenchymal stem cell injection with arthroscopic treatment in older patients with osteochondral lesions of the talus.</a> Am. J. Sports Med. 41 (5), 1090-1099 (2013).</li><li>Kim, Y. S., Lee, H. J., Choi, Y. J., Kim, Y. I., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+an+injection+of+a+stromal+vascular+fraction+containing+adipose-derived+mesenchymal+stem+cells+influence+the+outcomes+of+marrow+stimulation+in+osteochondral+lesions+of+the+talus?+A+clinical+and+magnetic+resonance+imaging+study.">Does an injection of a stromal vascular fraction containing adipose-derived mesenchymal stem cells influence the outcomes of marrow stimulation in osteochondral lesions of the talus? A clinical and magnetic resonance imaging study.</a> Am. J. Sports Med. 42 (10), 2424-2434 (2014).</li><li>Kim, Y. S., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injection+of+Mesenchymal+Stem+Cells+as+a+Supplementary+Strategy+of+Marrow+Stimulation+Improves+Cartilage+Regeneration+After+Lateral+Sliding+Calcaneal+Osteotomy+for+Varus+Ankle+Osteoarthritis:+Clinical+and+Second-Look+Arthroscopic+Results.">Injection of Mesenchymal Stem Cells as a Supplementary Strategy of Marrow Stimulation Improves Cartilage Regeneration After Lateral Sliding Calcaneal Osteotomy for Varus Ankle Osteoarthritis: Clinical and Second-Look Arthroscopic Results.</a> Arthroscopy. 32 (5), 878-889 (2016).</li><li>Kim, Y. S., Lee, M., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Additional+mesenchymal+stem+cell+injection+improves+the+outcomes+of+marrow+stimulation+combined+with+supramalleolar+osteotomy+in+varus+ankle+osteoarthritis:+short-term+clinical+results+with+second-look+arthroscopic+evaluation.">Additional mesenchymal stem cell injection improves the outcomes of marrow stimulation combined with supramalleolar osteotomy in varus ankle osteoarthritis: short-term clinical results with second-look arthroscopic evaluation.</a> J. Exp. Orthop. 3 (1), 12 (2016).</li><li>Hanke, C. W., Bernstein, G., Bullock, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+tumescent+liposuction+in+15,336+patients.+National+survey+results.">Safety of tumescent liposuction in 15,336 patients. National survey results.</a> Dermatol Surg. 21 (5), 459-462 (1995).</li><li>Illouz, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+liposuction.">Complications of liposuction.</a> Clin Plast Surg. 33 (1), 129-163 (2006).</li><li>Dixit, V. V., Wagh, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unfavourable+outcomes+of+liposuction+and+their+management.">Unfavourable outcomes of liposuction and their management.</a> Indian J Plast Surg. 46 (2), 377-392 (2013).</li><li>Lehnhardt, M., Homann, H. H., Daigeler, A., Hauser, J., Palka, P., Steinau, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Major+and+lethal+complications+of+liposuction:+review+of+72+cases+in+Germany+between+1998+and+2002.">Major and lethal complications of liposuction: review of 72 cases in Germany between 1998 and 2002.</a> Plast Reconstr Surg. 121 (6), 396e-403e (2008).</li><li>Usuelli, F. G., de Girolamo, L., Grassi, M., D'Ambrosi, R., Montrasio, U. A., Boga, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-Arthroscopic+Autologous+Matrix-Induced+Chondrogenesis+for+the+Treatment+of+Osteochondral+Lesions+of+the+Talus.">All-Arthroscopic Autologous Matrix-Induced Chondrogenesis for the Treatment of Osteochondral Lesions of the Talus.</a> Arthrosc Tech. 4 (3), e255-e259 (2015).</li><li>Simonson, D. C., Roukis, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+ankle+arthroscopy+for+the+treatment+of+anterolateral+soft-tissue+impingement.">Safety of ankle arthroscopy for the treatment of anterolateral soft-tissue impingement.</a> Arthroscopy. 30 (2), 256-259 (2014).</li><li>Suzangar, M., Rosenfeld, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ankle+arthroscopy:+is+preoperative+marking+of+the+superficial+peroneal+nerve+important?.">Ankle arthroscopy: is preoperative marking of the superficial peroneal nerve important?.</a> J. Foot. Ankle Surg. 51 (2), 179-181 (2012).</li><li>Kraeutler, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+Concepts+Review+Update:+Osteochondral+Lesions+of+the+Talus.">Current Concepts Review Update: Osteochondral Lesions of the Talus.</a> Foot Ankle Int. 38 (3), 331-342 (2017).</li><li>Looze, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+Management+of+Osteochondral+Lesions+of+the+Talus.">Evaluation and Management of Osteochondral Lesions of the Talus.</a> Cartilage. 8 (1), 19-30 (2017).</li><li>Dragoo, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Healing+full-thickness+cartilage+defects+using+adipose-derived+stem+cells.">Healing full-thickness cartilage defects using adipose-derived stem cells.</a> Tissue Eng. 13 (7), 1615-1621 (2007).</li><li>Lee, S. Y., Kim, W., Lim, C., Chung, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+Lateral+Epicondylosis+by+Using+Allogeneic+Adipose-Derived+Mesenchymal+Stem+Cells:+A+Pilot+Study.">Treatment of Lateral Epicondylosis by Using Allogeneic Adipose-Derived Mesenchymal Stem Cells: A Pilot Study.</a> Stem Cells. 33 (10), 2995-3005 (2015).</li><li>Feisst, V., Meidinger, S., Locke, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+bench+to+bedside:+use+of+human+adipose-derived+stem+cells.">From bench to bedside: use of human adipose-derived stem cells.</a> Stem Cells Cloning. 8, 149-162 (2015).</li></ol>

フェデリコ ・ ジュゼッペ ・ Usuelli、MD は、インテグラと Geistlich、助成金から個人的な費用と、ジマーから提出した応募作品以外の経費個人を報告します。

近年、臨床および前臨床試験は、別の筋骨格系疾患を治療するために ADSCs の効果に関する彼らの注意を集中しています。この記事の目的は、鏡視下手術 microperforations と microfractured と精製脂肪を使用して距骨骨軟骨病変の治療のためのプロトコルを説明します。プロトコルには、合併症の高いリスクのいくつかの重要な手順が含まれます。脂肪収穫時に合併症は、局所または全身合併症に分?...

関節鏡は、痛みを軽減する機能を復元して、特に若くてアクティブな患者の生活の質を向上させることを目的 (Olt) 距骨骨軟骨病変の治療のためのゴールド スタンダードです。
現在、鏡視下手術のテクニックは、3 つの方法で分類できます。修復の技術は、デブリドマンと病変のレベルで microperforations を介して骨髄由来の細胞を刺激します。再建の手法は、ostechondral の自家および異種移植片を使用して病変を置き換えられます。損傷した組織1,2,3,4,5,6 を再構築する多能性細胞を区別し、複製の機能を利用して再生技術.
近年、再生技術、Olt と特に間葉系幹細胞に由来する脂肪組織 (ADSCs)7,8の治療のため数多くの in vitroとin vivoの研究の対象となっています。,9. これらの間葉系幹細胞展示その他の組織から分離された他の多能性細胞に類似した形態的, 機能的特性彼らはまたいくつかに分化する能力を持っているし、別の細胞ラインのin vitroとin vivoの両方10、11,12,13.これらの細胞に関する研究の焦点局在の主因、組織13,14グラム当たり 5,000 細胞濃度正常体重の 30% から 10% を表し、実際。その一方で、これらの細胞の使用を制限する要因を検査中に、処理に関連します。脂肪細胞、コラーゲン線維と正常血管コンポーネントの集合体を含む lipoaspirate コラーゲン、タイプ I、培養前に、の溶血を受ける酵素によって処理されます。ここでの目的は、microfractured と精製脂肪を使用して距骨骨軟骨病変の治療のためのプロトコルを説明します。

<table><tbody><tr><td>PROCESS KIT - PROCESSING KIT FOR FAT TISSUE</td><td>LIPOGEMS</td><td>LG PK 60</td><td>Lipogems Kit to obtain microfractured and purified ADSCs</td></tr><tr><td>HINTERMANN SPREADER</td><td>INTEGRA</td><td>119654</td><td>The spreader allow to access most of the talar dome, in particular in case of posterior lesion</td></tr><tr><td>CUP CURETTE</td><td>ARTHREX</td><td>AR-8655-02</td><td>To remove the damaged cartilage and necrotic and sclerotic bone</td></tr><tr><td>CHONDRAL PICK 30&amp;deg; TIP / 60&amp;deg; TIP</td><td>ARTHREX</td><td>AR-8655-05&lt;br/&gt; AR-8655-06</td><td>To perfrom microperforation at the level of the lesion, stimulating bleeding from the subchondral bone</td></tr><tr><td>SHAVER</td><td>ARTHREX</td><td>AR-7300SR</td><td>To clean the joint and aspirate water</td></tr></tbody></table>

autologous microfractured and purified adipose tissue for arthroscopic management of osteochondral lesions of the talus

近年、再生技術がますます勉強し、距骨の軟骨病変を治療するために使用されます。特に、いくつかの研究は脂肪組織由来間葉系幹細胞に彼らの注意を焦点を当てています。脂肪由来幹細胞 (ADSCs) は形態学的特徴とその他の間葉系細胞に似た性質を示すし、数行細胞に分化することができます。また、これらの細胞は 5,000 細胞組織の 1 グラムあたりの濃度を正常体重の 10-30% を表す、皮下組織で広く利用可能ではあります。
紹介したテクニックで最初のステップは、腹部と微視破壊および浄化; のプロセスから ADSCs を収穫次に、手術の手順は、以下の軟部組織の解離より良い共同可視化、標準の開く手順と比較してより高速な回復と完全 arthroscopically されます。関節鏡は最初のフェーズは病変の識別、分離、microperforations; と準備によって特徴付けられる2 番目の手順では、実行のドライには、病変のレベルで脂肪の注入が含まれます。
2016 年 1 月、2016 年 9 月の間は、4 人の患者は microfractured と精製脂肪距骨骨軟骨病変の鏡視下手術の治療を受けた。すべての患者は報告された合併症と臨床症状の改善の手術後 6 ヶ月を報告します。最新のフォロー アップで機能のスコアは有望であり、信頼性の高い痛みの軽減と距骨の軟骨病変を有する患者の改善手法を提供することを確認します。

2016 年 1 月、2016 年 9 月の間は、4 人の患者は microfractured と精製脂肪距骨骨軟骨病変の鏡視下手術の治療を受けた。すべての患者は、手術後 6 ヶ月の臨床症状の改善を報告しました。表 1に予備的な臨床結果が報告されます。合併症は報告されませんでした。
近年では、足と足首の病態の治療のための ADSCs の?...

Watch this Scientific Journal Video about Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus at JoVE.com

Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

本研究の目的は、microfractured と精製脂肪由来幹細胞を使用して距骨骨軟骨病変の鏡視下手術治療のプロトコルを報告することです。

自己 Microfractured と距骨骨軟骨損傷の鏡視下手術の管理のための精製脂肪組織

In recent years, regenerative techniques have been increasingly studied and used to treat osteochondral lesions of the talus. In particular, several ...

autologous-microfractured-purified-adipose-tissue-for-arthroscopic

Research

JoVE Journal

Medicine

10.2K ビュー.  IRCCS Istituto Ortopedico Galeazzi - C.A.S.C.O. Piede e Caviglia. 本研究の目的は、microfractured と精製脂肪由来幹細胞を使用して距骨骨軟骨病変の鏡視下手術治療のプロトコルを報告することです。

ビデオ: Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. Adipose-derived stromal cells (ASCs) have proven to be an abundant source of multipotent stem cells capable of undergoing osteogenic, chondrogenic, adipogenic, and myogenic differentiation. Many studies have explored the osteogenic potential of these cells in vivo with the use of various scaffolding biomaterials for cellular delivery. It has been demonstrated that by utilizing an osteoconductive, hydroxyapatite-coated poly(lactic-co-glycolic acid) (HA-PLGA) scaffold seeded with ASCs, a critical-sized calvarial defect, a defect that is defined by its inability to undergo spontaneous healing over the lifetime of the animal, can be effectively show robust osseous regeneration. This in vivo model demonstrates the basis of translational approaches aimed to regenerate the bone tissue - the cellular component and biological matrix. This method serves as a model for the ultimate clinical application of a progenitor cell towards the repair of a specific tissue defect.

This protocol describes the isolation of adipose-derived stromal cells from lipoaspirate and the creation of a 4 mm critical-sized calvarial defect to evaluate skeletal regeneration.

吸引脂肪から採取した脂肪由来間質細胞を使用したクリティカル·サイズの頭蓋冠欠損モデルの修復

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. ...

医学

Treatment of Ankle Osteoarthritis with Total Ankle Replacement Through a Lateral Transfibular Approach

Total ankle replacement (TAR) is a valid option for the treatment of ankle osteoarthritis. The traditional surgical approach for TAR is the anterior approach. Recently, the lateral transfibular approach to the ankle has gained popularity since a new TAR implant was designed to be performed via this approach that results in an ideal visualization of the center of rotation of the ankle and curved resections that allow for sparing bone cuts. The aim of the present paper is to present our preoperative, operative, and postoperative protocols for the treatment of the osteoarthritis of the ankle with TAR via lateral approach. We present our preoperative clinical and radiographic protocol. In addition, we describe our surgical technique with some technical tips. Finally, we report our follow-up schedule that includes the collection of clinical, functional, and radiographic data. The results of this procedure are encouraging: TAR through a lateral transfibular approach provides reliable pain relief and improvements in functional outcomes in patients with ankle osteoarthritis.

We present our preoperative, operative, and postoperative protocols for the treatment of osteoarthritis of the ankle with total ankle replacement via lateral transfibular approach.

横 Transfibular アプローチによって総足首置換と足関節変形性関節症の治療

Total ankle replacement (TAR) is a valid option for the treatment of ankle osteoarthritis. The traditional surgical approach for TAR is the anterior ...

行動学

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated cartilage lesions lead to ongoing pain, negatively affect the quality of life and predispose for osteoarthritis. During the last decades, several surgical techniques have been developed to treat such lesions. However, until now it was not possible to achieve a full repair in terms of covering the defect with hyaline articular cartilage or of providing satisfactory long-term recovery 2-4. Therefore, articular cartilage injuries remain a prime target for regenerative techniques such as Tissue Engineering. In contrast to other surgical techniques, which often lead to the formation of fibrous or fibrocartilaginous tissue, Tissue Engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells. Recent developments opened up promising possibilities for regenerative cartilage therapies.
The first cell based approach for the treatment of full-thickness cartilage or osteochondral lesions was performed in 1994 by Lars Peterson and Mats Brittberg who pioneered clinical autologous chondrocyte implantation (ACI) 5. Today, the technique is clinically well-established for the treatment of large hyaline cartilage defects of the knee, maintaining good clinical results even 10 to 20 years after implantation 6. In recent years, the implantation of autologous chondrocytes underwent a rapid progression. The use of an artificial three-dimensional collagen-matrix on which cells are subsequently replanted became more and more popular 7-9.
MACT comprises of two surgical procedures: First, in order to collect chondrocytes, a cartilage biopsy needs to be performed from a non weight-bearing cartilage area of the knee joint. Then, chondrocytes are being extracted, purified and expanded to a sufficient cell number in vitro. Chondrocytes are then seeded onto a three-dimensional matrix and can subsequently be re-implanted. When preparing a tissue-engineered implant, proliferation rate and differentiation capacity are crucial for a successful tissue regeneration 10. The use of a three-dimensional matrix as a cell carrier is thought to support these cellular characteristics 11.
The following protocol will summarize and demonstrate a technique for the isolation of chondrocytes from cartilage biopsies, their proliferation in vitro and their seeding onto a 3D-matrix (Chondro-Gide, Geistlich Biomaterials, Wollhusen, Switzerland). Finally, the implantation of the cell-matrix-constructs into artificially created chondral defects of a rabbit's knee joint will be described. This technique can be used as an experimental setting for further experiments of cartilage repair.

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

ウサギモデルにおける軟骨欠陥の改造と修理のためのマトリックス支援自家軟骨細胞移植

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated ...

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular cartilage and subchondral bone in recent years led to increased attention to restoration of the entire osteochondral unit. In comparison to chondral lesions the regeneration of osteochondral defects is much more complex and a far greater surgical and therapeutic challenge. The damaged tissue does not only include the superficial cartilage layer but also the subchondral bone. For deep, osteochondral damage, as it occurs for example with osteochondrosis dissecans, the full thickness of the defect needs to be replaced to restore the joint surface 1. Eligible therapeutic procedures have to consider these two different tissues with their different intrinsic healing potential 2. In the last decades, several surgical treatment options have emerged and have already been clinically established 3-6.
Autologous or allogeneic osteochondral transplants consist of articular cartilage and subchondral bone and allow the replacement of the entire osteochondral unit. The defects are filled with cylindrical osteochondral grafts that aim to provide a congruent hyaline cartilage covered surface 3,7,8. Disadvantages are the limited amount of available grafts, donor site morbidity (for autologous transplants) and the incongruence of the surface; thereby the application of this method is especially limited for large defects.
New approaches in the field of tissue engineering opened up promising possibilities for regenerative osteochondral therapy. The implantation of autologous chondrocytes marked the first cell based biological approach for the treatment of full-thickness cartilage lesions and is now worldwide established with good clinical results even 10 to 20 years after implantation 9,10. However, to date, this technique is not suitable for the treatment of all types of lesions such as deep defects involving the subchondral bone 11.
The sandwich-technique combines bone grafting with current approaches in Tissue Engineering 5,6. This combination seems to be able to overcome the limitations seen in osteochondral grafts alone. After autologous bone grafting to the subchondral defect area, a membrane seeded with autologous chondrocytes is sutured above and facilitates to match the topology of the graft with the injured site. Of course, the previous bone reconstruction needs additional surgical time and often even an additional surgery. Moreover, to date, long-term data is missing 12.
Tissue Engineering without additional bone grafting aims to restore the complex structure and properties of native articular cartilage by chondrogenic and osteogenic potential of the transplanted cells. However, again, it is usually only the cartilage tissue that is more or less regenerated. Additional osteochondral damage needs a specific further treatment. In order to achieve a regeneration of the multilayered structure of osteochondral defects, three-dimensional tissue engineered products seeded with autologous/allogeneic cells might provide a good regeneration capacity 11.
Beside autologous chondrocytes, mesenchymal stem cells (MSC) seem to be an attractive alternative for the development of a full-thickness cartilage tissue. In numerous preclinical in vitro and in vivo studies, mesenchymal stem cells have displayed excellent tissue regeneration potential 13,14. The important advantage of mesenchymal stem cells especially for the treatment of osteochondral defects is that they have the capacity to differentiate in osteocytes as well as chondrocytes. Therefore, they potentially allow a multilayered regeneration of the defect.
In recent years, several scaffolds with osteochondral regenerative potential have therefore been developed and evaluated with promising preliminary results 1,15-18. Furthermore, fibrin glue as a cell carrier became one of the preferred techniques in experimental cartilage repair and has already successfully been used in several animal studies 19-21 and even first human trials 22.
The following protocol will demonstrate an experimental technique for isolating mesenchymal stem cells from a rabbit's bone marrow, for subsequent proliferation in cell culture and for preparing a standardized in vitro-model for fibrin-cell-clots. Finally, a technique for the implantation of pre-established fibrin-cell-clots into artificial osteochondral defects of the rabbit's knee joint will be described.

An experimental technique for the treatment of osteochondral defects in the rabbit's knee joint is described. The implantation of allogeneic mesenchymal stem cells into osteochondral defects provides a promising development in the field of tissue engineering. The preparation of fibrin-cell-clots in vitro offers a standardized method for implantation.

フィブリン塊における同種間葉系幹細胞の注入によってウサギの膝関節における骨軟骨欠損の治療

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

Application of Atomic Force Microscopy to Detect Early Osteoarthritis

Biomechanical properties of cells and tissues not only regulate their shape and function but are also crucial for maintaining their vitality. Changes in elasticity can propagate or trigger the onset of major diseases like cancer or osteoarthritis (OA). Atomic force microscopy (AFM) has emerged as a strong tool to qualitatively and quantitatively characterize the biomechanical properties of specific biological target structures on a microscopic scale, measuring forces in a range from as small as the piconewton to the micronewton. Biomechanical properties are of special importance in musculoskeletal tissues, which are subjected to high levels of strain. OA as a degenerative disease of the cartilage results in the disruption of the pericellular matrix (PCM) and the spatial rearrangement of the chondrocytes embedded in their extracellular matrix (ECM). Disruption in PCM and ECM has been associated with changes in the biomechanical properties of cartilage. In the present study we used AFM to quantify these changes in relation to the specific spatial pattern changes of the chondrocytes. With each pattern change, significant changes in elasticity were observed for both the PCM and ECM. Measuring the local elasticity thus allows for drawing direct conclusions about the degree of local tissue degeneration in OA.

We present a method to investigate early osteoarthritic changes at the cellular level in articular cartilage by using atomic force microscopy (AFM).

早期変形性関節症の検出に対する原子間力顕微鏡の応用

Biomechanical properties of cells and tissues not only regulate their shape and function but are also crucial for maintaining their vitality. Changes in ...

生物工学

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

潜在的な軟骨再生のための脂肪組織由来間質血管分数の生産の臨床プロトコル

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

<meta charset="utf8"></head><body>外植片モデルを用いた骨形成のマルチモーダル解析

Comprehensive Characterization of Tissue Mineralization in an Ex Vivo Model

The extensive characterization of tissue mineralization in the context of bone regeneration represents a significant challenge, given the numerous modalities that are currently available for analysis. Here, we propose a workflow for a comprehensive evaluation of new bone formation using a relevant large animal osseous ex vivo explant. A bone defect (diameter = 3.75 mm; depth = 5.0 mm) is created in an explanted sheep femoral head and injected with a macroporous bone substitute loaded with a pro-osteogenic growth factor (bone morphogenetic protein 2 - BMP2). Subsequently, the explant is maintained in culture for a 28-day period, allowing cellular colonization and subsequent bone formation. To evaluate the quality and structure of newly mineralized tissue, the following successive methods are set up: (i) Characterization and high-resolution 3D images of the entire explant using micro-CT, followed by deep learning image analyses to enhance the discrimination of mineralized tissues; (ii) Nano-indentation to determine the mechanical properties of the newly formed tissue; (iii) Histological examinations, such as Hematoxylin/Eosin/Saffron (HES), Goldner's trichrome, and Movat's pentachrome to provide a qualitative assessment of mineralized tissue, particularly with regard to the visualization of the osteoid barrier and the presence of bone cells; (iv) Back-scattering scanning electron microscopy (SEM) mapping with internal reference to quantify the degree of mineralization and provide detailed insights into surface morphology, mineral composition, and bone-biomaterial interface; (v) Raman spectroscopy to characterize the molecular composition of the mineralized matrix and to provide insights into the persistence of BMP2 within the cement through the detection of peptide bonds. This multimodal analysis will provide an effective assessment of newly formed bone and comprehensive qualitative and quantitative insights into mineralized tissues. Through the standardization of these protocols, we aim to facilitate interstudy comparisons and improve the validity and reliability of research findings.

<meta charset="utf8"></head><body>著者スポットライト:骨再生研究における組織石灰化の特性評価のための高度な技術

The proposed protocol entails a global approach to assess bone formation in the context of bone regeneration using multimodal analyses. It aims to provide qualitative and quantitative information on new bone formation, enhancing the rigor and validity of basic and pre-clinical investigations.

Ex Vivoモデルにおける組織石灰化の包括的な特性評価

The extensive characterization of tissue mineralization in the context of bone regeneration represents a significant challenge, given the numerous ...

Destabilization of the Medial Meniscus and Cartilage Scratch Murine Model of Accelerated Osteoarthritis

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are used to mimic many aspects of the disease. The present protocol describes the destabilization and cartilage scratch model (DCS) of post-traumatic osteoarthritis. Based on the widely used destabilization of the medial meniscus (DMM) model, DCS introduces three scratches on the cartilage surface. The current article highlights the steps to destabilize the knee by transecting the medial meniscotibial ligament followed by three intentional superficial scratches on the articular cartilage. The possible analysis methods by dynamic weight-bearing, microcomputed tomography, and histology are also demonstrated. While the DCS model is not recommended for studies that focus on the effect of osteoarthritis on the cartilage, it enables the study of osteoarthritis development in a shorter time window, with special focus on (1) osteophyte formation, (2) osteoarthritic and injury pain, and (3) the effect of cartilage damage in the whole joint.

The present protocol describes the controlled microblade scratches on the surface of the articular cartilage after destabilizing the mouse knee by cutting the medial miniscotibial ligament. This animal model presents an accelerated form of osteoarthritis (OA) suitable for studying osteophyte formation, osteosclerosis, and early-stage pain.

加速変形性関節症の内側半月板および軟骨スクラッチマウスモデルの不安定化

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are ...

自己 Microfractured と距骨骨軟骨損傷の鏡視下手術の管理のための精製脂肪組織

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吸引脂肪から採取した脂肪由来間質細胞を使用したクリティカル·サイズの頭蓋冠欠損モデルの修復

横 Transfibular アプローチによって総足首置換と足関節変形性関節症の治療

ウサギモデルにおける軟骨欠陥の改造と修理のためのマトリックス支援自家軟骨細胞移植

フィブリン塊における同種間葉系幹細胞の注入によってウサギの膝関節における骨軟骨欠損の治療

早期変形性関節症の検出に対する原子間力顕微鏡の応用

潜在的な軟骨再生のための脂肪組織由来間質血管分数の生産の臨床プロトコル

Ex Vivoモデルにおける組織石灰化の包括的な特性評価

Ex Vivoモデルにおける組織石灰化の包括的な特性評価

Ex Vivoモデルにおける組織石灰化の包括的な特性評価

加速変形性関節症の内側半月板および軟骨スクラッチマウスモデルの不安定化