This research was supported by the FP7 European Research Council Grant 281501, ENGVASC.

<ol>
	<li>Engelsman, A. F., van der Mei, H. C., Ploeg, R. J., Busscher, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+phenomenon+of+infection+with+abdominal+wall+reconstruction.">The phenomenon of infection with abdominal wall reconstruction.</a> Biomaterials. 28 (14), 2314-2327 (2007).</li><li>De Coppi, P., 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=Myoblast-acellular+skeletal+muscle+matrix+constructs+guarantee+a+long-term+repair+of+experimental+full-thickness+abdominal+wall+defects.">Myoblast-acellular skeletal muscle matrix constructs guarantee a long-term repair of experimental full-thickness abdominal wall defects.</a> Tissue Eng. 12 (7), 1929-1936 (2006).</li><li>Shi, C., 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=Regeneration+of+full-thickness+abdominal+wall+defects+in+rats+using+collagen+scaffolds+loaded+with+collagen-binding+basic+fibroblast+growth+factor.">Regeneration of full-thickness abdominal wall defects in rats using collagen scaffolds loaded with collagen-binding basic fibroblast growth factor.</a> Biomaterials. 32 (3), 753-759 (2011).</li><li>Yezhelyev, M. V., Deigni, O., Losken, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+full-thickness+abdominal+wall+defects+following+tumor+resection.">Management of full-thickness abdominal wall defects following tumor resection.</a> Ann Plast Surg. 69 (2), 186-191 (2012).</li><li>Macchiarini, P., Walles, T., Biancosino, C., Mertsching, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+human+transplantation+of+a+bioengineered+airway+tissue.">First human transplantation of a bioengineered airway tissue.</a> J Thorac Cardiovasc Surg. 128 (4), 638-641 (2004).</li><li>Macchiarini, P., 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=Clinical+transplantation+of+a+tissue-engineered+airway.">Clinical transplantation of a tissue-engineered airway.</a> Lancet. 372 (9665), 2023-2030 (2008).</li><li>Atala, A., Bauer, S. B., Soker, S., Yoo, J. J., Retik, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-engineered+autologous+bladders+for+patients+needing+cystoplasty.">Tissue-engineered autologous bladders for patients needing cystoplasty.</a> Lancet. 367 (9518), 1241-1246 (2006).</li><li>Nishida, K., 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=Corneal+reconstruction+with+tissue-engineered+cell+sheets+composed+of+autologous+oral+mucosal+epithelium.">Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium.</a> N Engl J Med. 351 (12), 1187-1196 (2004).</li><li>Petite, H., 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=Tissue-engineered+bone+regeneration.">Tissue-engineered bone regeneration.</a> Nat Biotechnol. 18 (9), 959-963 (2000).</li><li>Banta, M. N., Kirsner, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulating+diseased+skin+with+tissue+engineering:+actinic+purpura+treated+with+Apligraf.">Modulating diseased skin with tissue engineering: actinic purpura treated with Apligraf.</a> Dermatol Surg. 28 (12), 1103-1106 (2002).</li><li>Vunjak-Novakovic, G., 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=Challenges+in+cardiac+tissue+engineering.">Challenges in cardiac tissue engineering.</a> Tissue engineering. Part B, Reviews. 16 (2), 169-187 (2010).</li><li>Novosel, E. C., Kleinhans, C., Kluger, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascularization+is+the+key+challenge+in+tissue+engineering.">Vascularization is the key challenge in tissue engineering.</a> Advanced Drug Delivery Reviews. 63 (4-5), 300-311 (2011).</li><li>Lesman, A., Gepstein, L., Levenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascularization+shaping+the+heart.">Vascularization shaping the heart.</a> Ann N Y Acad Sci. 1188, 46-51 (2010).</li><li>Patton Jr, H., Berry, S., Kralovich, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+human+acellular+dermal+matrix+in+complex+and+contaminated+abdominal+wall+reconstructions.">Use of human acellular dermal matrix in complex and contaminated abdominal wall reconstructions.</a> The Am J of Surg. 193 (3), 360-363 (2007).</li><li>Menon, N. G., 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=Revascularization+of+human+acellular+dermis+in+full-thickness+abdominal+wall+reconstruction+in+the+rabbit+model.">Revascularization of human acellular dermis in full-thickness abdominal wall reconstruction in the rabbit model.</a> Ann Plast Surg. 50 (5), 523-527 (2003).</li><li>Buinewicz, B., Rosen, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acellular+cadaveric+dermis+(AlloDerm):+a+new+alternative+for+abdominal+hernia+repair.">Acellular cadaveric dermis (AlloDerm): a new alternative for abdominal hernia repair.</a> Ann Plast Surg. 52 (2), 188-194 (2004).</li><li>Bringman, S., 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=Hernia+repair:+the+search+for+ideal+meshes.">Hernia repair: the search for ideal meshes.</a> Hernia. 14 (1), 81-87 (2010).</li><li>Meintjes, J., Yan, S., Zhou, L., Zheng, S., Zheng, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthetic+biological+and+composite+scaffolds+for+abdominal+wall+reconstruction.">Synthetic biological and composite scaffolds for abdominal wall reconstruction.</a> Exp rev of med dev. 8 (2), 275-288 (2011).</li><li>Cheng, G., 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=Engineered+blood+vessel+networks+connect+to+host+vasculature+via+wrapping-and-tapping+anastomosis.">Engineered blood vessel networks connect to host vasculature via wrapping-and-tapping anastomosis.</a> Blood. 118 (17), 4740-4749 (2011).</li><li>Shandalov, Y., 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=An+engineered+muscle+flap+for+reconstruction+of+large+soft+tissue+defects.">An engineered muscle flap for reconstruction of large soft tissue defects.</a> PNAS of the USA. 111 (16), 6010-6015 (2014).</li><li>Zhang, T. Y., Suen, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+fast+parallel+algorithm+for+thinning+digital+patterns.">A fast parallel algorithm for thinning digital patterns.</a> Commun. ACM. 27 (3), 236-239 (1984).</li><li>Luna, L. G., Luna, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Manual of Histo Stain Meth ; of the Arm Forcs Inst of Path. , (1968).</li><li>Choi, J. H., 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=Adipose+tissue+engineering+for+soft+tissue+regeneration.">Adipose tissue engineering for soft tissue regeneration.</a> Tissue engineering. Part B, Reviews. 16 (4), 413-426 (2010).</li><li>Bellows, C. F., Alder, A., Helton, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abdominal+wall+reconstruction+using+biological+tissue+grafts:+present+status+and+future+opportunities.">Abdominal wall reconstruction using biological tissue grafts: present status and future opportunities.</a> Exp rev of med dev. 3 (5), 657-675 (2006).</li><li>Caspi, O., 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=Tissue+engineering+of+vascularized+cardiac+muscle+from+human+embryonic+stem+cells.">Tissue engineering of vascularized cardiac muscle from human embryonic stem cells.</a> Circ Res. 100 (2), 263-272 (2007).</li><li>Kaufman-Francis, K., Koffler, J., Weinberg, N., Dor, Y., Levenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineered+vascular+beds+provide+key+signals+to+pancreatic+hormone-producing+cells.">Engineered vascular beds provide key signals to pancreatic hormone-producing cells.</a> PloS one. 7 (7), e40741 (2012).</li><li>Kaully, T., Kaufman-Francis, K., Lesman, A., Levenberg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascularization--the+conduit+to+viable+engineered+tissues.">Vascularization--the conduit to viable engineered tissues.</a> Tiss eng. Part B, Reviews. 15 (2), 159-169 (2009).</li><li>Koffler, 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=Improved+vascular+organization+enhances+functional+integration+of+engineered+skeletal+muscle+grafts.">Improved vascular organization enhances functional integration of engineered skeletal muscle grafts.</a> PNAS of the USA. 108 (36), 14789-14794 (2011).</li><li>Lesman, 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=Transplantation+of+a+tissue-engineered+human+vascularized+cardiac+muscle.">Transplantation of a tissue-engineered human vascularized cardiac muscle.</a> Tisseng. Part A. 16 (1), 115-125 (2010).</li><li>Levenberg, S., 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=Engineering+vascularized+skeletal+muscle+tissue.">Engineering vascularized skeletal muscle tissue.</a> Nat Biotechnol. 23 (7), 879-884 (2005).</li><li>Bearzi, C., 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=PlGF-MMP9-engineered+iPS+cells+supported+on+a+PEG-fibrinogen+hydrogel+scaffold+possess+an+enhanced+capacity+to+repair+damaged+myocardium.">PlGF-MMP9-engineered iPS cells supported on a PEG-fibrinogen hydrogel scaffold possess an enhanced capacity to repair damaged myocardium.</a> Cell death &amp; disease. 5, e1053 (2014).</li><li>Zhang, M., 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=SDF-1+expression+by+mesenchymal+stem+cells+results+in+trophic+support+of+cardiac+myocytes+after+myocardial+infarction.">SDF-1 expression by mesenchymal stem cells results in trophic support of cardiac myocytes after myocardial infarction.</a> FASEB J : official publication of the .Fed Am Soc Exp Biol. 21 (12), 3197-3207 (2007).</li><li>Dvir, T., 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=Prevascularization+of+cardiac+patch+on+the+omentum+improves+its+therapeutic+outcome.">Prevascularization of cardiac patch on the omentum improves its therapeutic outcome.</a> PNAS. 106 (35), 14990-14995 (2009).</li><li>Marsano, 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=The+effect+of+controlled+expression+of+VEGF+by+transduced+myoblasts+in+a+cardiac+patch+on+vascularization+in+a+mouse+model+of+myocardial+infarction.">The effect of controlled expression of VEGF by transduced myoblasts in a cardiac patch on vascularization in a mouse model of myocardial infarction.</a> Biomaterials. 34 (2), 393-401 (2013).</li><li>Rufaihah, A. 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=Enhanced+infarct+stabilization+and+neovascularization+mediated+by+VEGF-loaded+PEGylated+fibrinogen+hydrogel+in+a+rodent+myocardial+infarction+model.">Enhanced infarct stabilization and neovascularization mediated by VEGF-loaded PEGylated fibrinogen hydrogel in a rodent myocardial infarction model.</a> Biomaterials. 34 (33), 8195-8202 (2013).</li><li>Nillesen, S. T. M., 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=Increased+angiogenesis+in+acellular+scaffolds+by+combined+release+of+FGF2+and+VEGF.">Increased angiogenesis in acellular scaffolds by combined release of FGF2 and VEGF.</a> J of Contr Release. 116 (2), e88-e90 (2006).</li><li>Sekine, H., 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=In+vitro+fabrication+of+functional+three-dimensional+tissues+with+perfusable+blood+vessels.">In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels.</a> Nat Commun. 4, 1399 (2013).</li><li>Tee, R., 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=Transplantation+of+engineered+cardiac+muscle+flaps+in+syngeneic+rats.">Transplantation of engineered cardiac muscle flaps in syngeneic rats.</a> Tiss eng. Part A. (19-20), 1992-1999 (2012).</li><li>Morritt, A. N., 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=Cardiac+tissue+engineering+in+an+in+vivo+vascularized+chamber.">Cardiac tissue engineering in an in vivo vascularized chamber.</a> Circulation. 115 (3), 353-360 (2007).</li></ol>

The authors declare that they have no competing financial interests.

وقد التقى التقدم في هندسة الأنسجة مع الطلب المتزايد على الأنسجة بديلا عن إعادة الإعمار من أنواع الأنسجة المختلفة. وقد تم تقييم مجموعة متنوعة من الاصطناعية 1،17،18 والبيولوجية 14-16 المواد وكذلك أساليب التصنيع لقدرتها على التصدي لهذه المطالب. ومع ذلك، على الرغم من التقدم في مجال الرعاية السريرية وهندسة الأنسجة، واستعادة عيوب جدار البطن كامل سماكة لا تزال تشكل تحديا. يجب أن تكون الأنسجة كافية لإعادة بناء مثل هذه العيوب الضخمة (1) سميكة و(2) أوعية دموية، وإظهار (3) السلامة الميكانيكية، و (4) الجدوى مع مرور الوقت 23. هنا، نقدم بروتوكول خطوة بخطوة مفصل لإنشاء سميكة، وقابلة للحياة، وأوعية دموية رفرف الأنسجة، والتي يمكن أن تكون بمثابة بديل لاللوحات النسيج ذاتي. شيد رفرف ملفقة في خطوتين: (1) تم زرعها سقالة PLLA / PLGA حول الأوعية AV مذكرة التفاهمحد ذاته hindlimb وفصلها عن الأنسجة المحيطة بها للسماح الأوعية الدموية عن طريق الأوعية AV فقط. (2) ثم نقل خلق أوعية دموية، رفرف نسيج سميك مع السفن AV، الذي شغل منصب عنيق لها، إلى وجود خلل جدار البطن كامل سماكة. رفرف السليم الأوعية الدموية ضروري لنجاح التكامل في غضون 17،18،24 المضيف. الطرق المختلفة لخلق الأنسجة المهندسة أوعية دموية من أجل تحسين امدادات الاوكسجين ونشرها في الأنسجة السميكة، وقد نوقشت في الأدب. ومن بين هذه الأساليب التي تنطوي على زرع الخلايا البطانية (ECS) على أنواع مختلفة سقالة 13،20،25-30، تقنيات مختلفة لتوفير عوامل عائية إلى الموقع زرع (إما عن طريق الإدارة مباشرة عن طريق الحقن، واستخدام الخلايا تحولت يعبرون عن العوامل 31 الإفراج -34 أو بطيئة من السقالات مختلفة 35،36)، واستخدام المفاعلات الحيوية لضمان الأنسجة المهندسة نضح 37 </سوب&gt; وتوظيف غرف حلقة AV 38،39. هنا، كان أوعية دموية الكسب غير المشروع الأنسجة في الجسم الحي، من خلال استغلال السفن ذاتي. الكسب غير المشروع، والتي أثبتت قابلة للحياة، وأوعية دموية perfused، ثم تم نقلها كما رفرف نسيج سميك لإصلاح الخلل جدار البطن كامل سماكة. كان أسبوع واحد بعد نقل-رفرف قابلة للحياة وظهرت قوة ميكانيكية كافية لدعم أحشاء البطن. هنا، كنا الفئران عارية athymic، التي تحمل نظام المناعة وضعف. عند استخدام أي نوع آخر من الماوس أو حيوان آخر، ينبغي أن تؤخذ في إمكانية التفاعلات المناعية في الاعتبار (وخصوصا عندما بذر السقالات مع الخلايا). وتشمل القيود الأخرى لهذه التقنية (1) نقل السفن AV الفخذ، التي تزود تدفق الدم إلى hindlimb. ومع ذلك، فإن تقنية يترك العميقة والأوعية الفخذية العميقة دون أن تمس ولم يلاحظ وجود نقص التروية hindlimb بعد نقل رفرف. وعلاوة على ذلك، في الحيوانات الكبيرة والإنسان، واستخداموسيتم إبلاغ الأوعية الطرفية لأنها زائدة في الجسم. (2) الوقت اللازم لتحقيق الأوعية الدموية كاف قبل رفرف نقل، يمكن أن يكون عاملا محددا في الحالات العاجلة. و (3) يتم تنفيذ إنشاء رفرف في الجسم الحي، والتي يمكن أن تحد من أهميتها في البشر. في المستقبل، ونحن نهدف إلى تطوير وسيلة لخلق هذا رفرف خارج الحي. الاستفادة من طريقة عرض تكمن في القدرة على تعزيز تشكيل الأنسجة ذاتي (مع خلايا ذاتي) حول السقالة. تقدم الأنسجة الناتجة بديل مناسب لرفرف نسيج ذاتي. وعلاوة على ذلك، يمكن المصنف السقالة مع الخلايا ذاتي قبل الزرع، لزيادة تحسين الأوعية الدموية الكسب غير المشروع والتكامل داخل النسيج المضيف. استخدام الخلايا ذاتي وسقالة القابلة للتحلل وحيويا، يمكن التحايل على خطر كبير من التفاعلات المناعية الرفض ويقدم بديلا لاللوحات ذاتي مع تجنب الحصاد وscarifi بعد العملية الجراحيةالأيونات الموجبة. الطريقة الموصوفة يمكن أن تترجم إلى التجارب في الحيوانات الكبيرة وزيادة توسيع إلى التجارب السريرية على البشر. وعلاوة على ذلك يمكن أن يترجم إلى إصلاح الأنسجة التالفة المختلفة داخل الجسم. في البشر والحيوانات في كبيرة، يمكن أن تتولد اللوحات حول الأوعية المحيطية بدلا من الأوعية AV الفخذ.

كثيرا ما تنشأ عيوب جدار البطن بعد الصدمة شديدة، علاج السرطان، والحروق وإزالة شبكة المصابة. هذه العيوب غالبا ما تنطوي على فقدان الأنسجة الهامة، والتي تتطلب عمليات جراحية معقدة وتمثل تحديا كبيرا للجراحين إعادة الإعمار البلاستيك 1-4. الباحثين هندسة الأنسجة البحث عن مصادر جديدة للأنسجة اصطناعية واستكشفت مواد مختلفة، ومصادر الخلايا وعوامل النمو. وقد ذكرت سابقا ترميم ناجحة من الأنسجة المختلفة، مثل 5،6 القصبة الهوائية، المثانة 7، 8 القرنية والعظام والجلد 9 10، عن طريق زرع الأنسجة المهندسة. ومع ذلك، تصنيع الأنسجة أوعية دموية سميكة هندسيا، وخاصة لإعادة الإعمار من عيوب كبيرة، لا يزال يشكل تحديا كبيرا في هندسة الأنسجة. واحدة من العوامل الرئيسية التي تحد من سمك بناء أنسجة قابلة للحياة هو السيطرة على امدادات الاوكسجين الى سلبيات لهاخلايا tituent. عند الاعتماد على نشرها، بناء سمك يقتصر على أن من طبقة رقيقة جدا. المسافة القصوى بين الأكسجين والشعيرات الدموية-توريد المغذيات في الجسم الحي حوالي 200 ميكرون، الذي يرتبط مع الحد نشر الأكسجين 11،12. الأوعية الدموية غير كافية يمكن أن يؤدي إلى نقص تروية الأنسجة وتتصاعد لارتشاف الأنسجة أو نخر 13. وبالإضافة إلى ذلك، يجب أن تكون المادة المثالية تستخدم لإعادة بناء الأنسجة حيويا وغير مناعة. كما يجب أن تكون قادرة على تعزيز مزيد من التكامل من الخلايا المضيفة مع مادة بيولوجية، والحفاظ على السلامة الهيكلية. تم مختلف البيولوجية 14-16 والاصطناعية 1،17،18 المصفوفات استكشاف سابقا لإعادة الإعمار الأنسجة، ولكن استخدامها لا يزال محدودا بسبب عدم وصول الدم فعالة، والتهابات أو عدم كفاية قوة الأنسجة. في هذه الدراسة، وحيويا، خلية الإدارة الانتخابيةكان مزروع سقالة edded تتألف من إدارة الغذاء والدواء (FDA) بولي وافق عليها-L حمض اللاكتيك (PLLA) / بولي حامض اللبنيك في-CO-الجليكوليك (PLGA)، وحول الشريان الفخذي والوريد (AV) سفن ماوس عارية و فصلها عن الأنسجة المحيطة بها، وضمان الأوعية الدموية من أوعية AV فقط. ، وكان الكسب غير المشروع أسبوع واحد بعد زرع قابل للحياة، سميكة وأوعية دموية بشكل جيد. هذا النسيج أوعية دموية سميكة مع السفن AV، ثم تم نقلها كما رفرف pedicled إلى البطن عيب كامل سمك في نفس الماوس. كان رفرف أسبوع واحد بعد نقل قابل للحياة، أوعية دموية ودمجها بشكل جيد مع الأنسجة المحيطة بها، مع ما يكفي من القوة لدعم أحشاء البطن. وهكذا، فإن سميكة، ورفرف الأنسجة أوعية دموية هندسيا، واضعة في عنيق ذاتي، ويقدم طريقة جديدة لإصلاح عيوب جدار البطن كامل سماكة.

<table><tbody><tr><td>small fine straight scissors</td><td>Fine Science Tools (FST)</td><td>14090-09</td><td></td></tr><tr><td>spring scissors</td><td>Fine Science Tools (FST)</td><td>15003-08</td><td></td></tr><tr><td>straight forceps with fine tip</td><td>Fine Science Tools (FST)</td><td>11251-20</td><td></td></tr><tr><td>serrated forceps&amp;nbsp;</td><td>Fine Science Tools (FST)</td><td>11050-10</td><td></td></tr><tr><td>needle holder</td><td>Fine Science Tools (FST)</td><td>12500-12</td><td></td></tr><tr><td>Small vessel cauterizer&amp;nbsp;</td><td>Fine Science Tools (FST)</td><td>18000-00</td><td></td></tr><tr><td>Duratears</td><td>Alcon</td><td>5686</td><td></td></tr><tr><td>Sedaxylan</td><td>Euravet</td><td>DJ03</td><td></td></tr><tr><td>Clorketam 1000</td><td>Vetoquinol</td><td>4A0726B</td><td></td></tr><tr><td>Buprenorphine</td><td>vetmarket</td><td>B15100</td><td></td></tr><tr><td>4-0 silk sutures</td><td>Assut sutures</td><td>647</td><td></td></tr><tr><td>6-0 polypropylene sutures</td><td>Assut sutures</td><td>9351F</td><td></td></tr><tr><td>8-0 silk sutures</td><td>Assut sutures</td><td>684568</td><td></td></tr><tr><td>Insulin syringe (6 mm needle)</td><td>BD</td><td>324911</td><td></td></tr><tr><td>Vevo 2100 high-resolution ultrasound system</td><td>VisualSonics inc.</td><td></td><td></td></tr><tr><td>MS250 non-linear transducer</td><td>VisualSonics inc.</td><td></td><td></td></tr><tr><td>Micromarker non-targeted contrast agent</td><td>VisualSonics inc.</td><td>VS-11694</td><td></td></tr><tr><td>tail vein catheter</td><td>VisualSonics inc.</td><td>VS-11912</td><td></td></tr><tr><td>Vevo 2100 software</td><td>VisualSonics inc.</td><td></td><td></td></tr><tr><td>fluorescein isothiocyanate-conjugated dextran</td><td>Sigma</td><td>FD500S</td><td></td></tr><tr><td>Matlab</td><td>Mathworks, MA, USA</td><td></td><td></td></tr><tr><td>Kimwipes</td><td>Kimtech</td><td>34120</td><td></td></tr><tr><td>antigen unmasking solution</td><td>Vector laboratories</td><td>H-3300</td><td></td></tr><tr><td>anti-CD31 antibody</td><td>Abcam&amp;nbsp;</td><td>ab28364</td><td></td></tr><tr><td>biotinylated goat anti-rabbit (secondary) antibody</td><td>Vector laboratories</td><td>BA-1000</td><td></td></tr><tr><td>streptavidin-peroxidase</td><td>Jackson&amp;nbsp;</td><td>016-030-084</td><td></td></tr><tr><td>Mayer&#039;s hamatoxylin solution</td><td>Sigma-Aldrich</td><td>MHS-16</td><td></td></tr><tr><td>aminoethylcarbazole (AEC) substrate kit</td><td>Life technologies, Invitrogen&amp;nbsp;</td><td>00-2007</td><td></td></tr><tr><td>Vectamount</td><td>Vector laboratories</td><td>H-5501</td><td></td></tr></tbody></table>

engineered vascularized muscle flap

One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen supply to the cell microenvironment, as passive diffusion is limited to a very thin layer. Although various materials have been described to restore the integrity of full-thickness defects of the abdominal wall, no material has yet proved to be optimal, due to low graft vascularization, tissue rejection, infection, or inadequate mechanical properties. This protocol describes a means of engineering a fully vascularized flap, with a thickness relevant for muscle tissue reconstruction. Cell-embedded poly L-lactic acid/poly lactic-co-glycolic acid constructs are implanted around the mouse femoral artery and vein and maintained in vivo for a period of one or two weeks. The vascularized graft is then transferred as a flap towards a full thickness defect made in the abdomen. This technique replaces the need for autologous tissue sacrifications and may enable the use of in vitro engineered vascularized flaps in many surgical applications.

الكسب غير المشروع الأوعية الدموية ونضح في الجسم الحي تم زرع الطعوم واحد أو أسبوعين قبل نقلهم كما اللوحات المحورية. في أسبوع وأسبوعين بعد الزرع، والمراقبة الجسيمة لمنطقة الكسب غير المشروع كشفت ترقيع الأنسجة القابلة للحياة وأوعية دموية. هذه الطعوم ثبت أن أوعية دموية للغاية، على النحو الذي يحدده المناعية CD31 إيجابي (الشكل 1A)، وperfused للغاية، كما يتضح من القياسات حقن الوريد الذيل والموجات فوق الصوتية FITC-ديكستران. وقد لوحظت العديد من السفن بالفعل في أسبوع واحد بعد الزرع، وهو الرقم الذي ارتفع بشكل ملحوظ بعد أسبوع إضافي في محيط الأوعية AV. أظهر تقرير ديكستران المستندة FITC لكثافة السفينة وظيفية (FVD) (الشكل 1B) أن الكسب غير المشروع وأوعية دموية للغاية في أسبوع واحد postimplantation والتي لم تتخذ أي تغييرات هامة في أسبوع إضافي في الجسم الحي. وقد أثبتت المباح السفينة ونضحبواسطة التصوير بالموجات فوق الصوتية (كان FVD 5.71 مم - 1 ± 0.51 ملم - 1 و 10.28 مم - 1 ± 2.71 ملم - 1، أسبوع وأسبوعين بعد الزرع، على التوالي) أكد الشكل 1C المضيف الفخذ المباح السفينة وسلامة بعد الكسب غير المشروع الزرع. . وعلاوة على ذلك، كشف الفحص بالأشعة فوق الصوتية نضح داخل منطقة الكسب غير المشروع، والذي كان أعلى قليلا أسبوعين بعد الزرع بالمقارنة مع أسبوع واحد بعد زرع (1D الشكل). خصائص رفرف الفحص الإجمالي من اللوحات أسبوع واحد بعد نقل-كشف أنسجة قابلة للحياة، أوعية دموية ومتكاملة بشكل جيد. خضعت اللوحات الالتزام الراسخ محيطهم. عند مقارنة، اللوحات جزءا لا يتجزأ من الخلية قبل أوعية دموية للسيطرة على اخلوي، الطعوم nonvascularized، أظهر السابق الخصائص الميكانيكية الفائقة، كما يتضح من زيادة صلابة وقوة ( <stroنانوغرام&gt; الشكل 2). نحن أيضا لاحظت أن تفزر الجرح وحدث فتق أقل كثيرا في الفئران التي عولجت مع اللوحات جزءا لا يتجزأ من الخلية، بالمقارنة مع الحيوانات تعامل مع الطعوم السيطرة، والتي يمكن أن يعزى إلى زيادة القوة الميكانيكية للأنسجة المزروعة. <img alt="الشكل 1" src="/files/ftp_upload/52984/52984fig1.jpg" /> الشكل 1. الكسب غير المشروع الأوعية الدموية. (A) كثافة السفن CD31 إيجابي، قاس في أسبوع وأسبوعين postimplantation مقارنة بالمجموعة الضابطة. كل القيم هي طبيعية إلى منطقة الكسب غير المشروع (مم 2). * = T-اختبار ف &lt;0.05. (B) وظيفية كثافة الأوعية الدموية (FVD) في أسبوع وأسبوعين postimplantation. لم يلاحظ أي اختلاف كبير، ص = 0.08 (T-اختبار). (C) سفن براءات AV كما تصويرها باستخدام الموجات فوق الصوتية في وضع لون دوبلر. الأزرق وتمثيلا الحمراءر تدفق الدم في الكسب غير المشروع. ويبين رباعي الأصفر منطقة الكسب غير المشروع. (D) الكسب غير المشروع نضح في واحد وأسبوعين postimplantation. (E و F) H &amp; E تلطيخ من اللوحات المتكاملة مع الأنسجة المضيف: نقطة السهم الأسود الشريان قابلة للحياة. السهام البيضاء تشير ألياف العضلات في منطقة البطن. الأسهم الصفراء تشير بقايا سقالة. (H وG) توزيع ديكستران FITC كما يتضح من الصور المجهري مبائر. لجميع قرارات، كان حجم العينة ن ≥ 3 ويتم تمثيل كل القيم كما يعني الخطأ ± المعياري للمتوسط. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52984/52984fig1large.jpg" target="_blank">الرجاء انقر هنا لعرض نسخة أكبر من هذا الرقم.</a> <img alt="الرقم 2" src="/files/ftp_upload/52984/52984fig2.jpg" /> الرقم 2. الخواص الميكانيكية للرفرف أسبوع واحدبعد النقل. (A) وتصلب و(B) قوة الشد في نهاية المطاف (UTS) من اللوحات. السيطرة تقف والكسب غير المشروع فارغة دون الخلايا، خلايا تقف الطعوم الثقافة الثلاثية. * = T-اختبار ف &lt;0.05، ن = تتمثل 3. القيم كما يعني الخطأ ± المعياري للمتوسط. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/52984/52984fig2large.jpg" target="_blank">الرجاء انقر هنا لعرض نسخة أكبر من هذا الرقم.</a>

Watch this Scientific Journal Video about Engineered Vascularized Muscle Flap at JoVE.com

Engineered Vascularized Muscle Flap

To date, thick tissue defects are typically reconstructed by applying autologous tissue flaps or engineered tissues. In this protocol, we present a new method for engineering vascularized tissue flap bearing an autologous pedicle, to serve as a substitute to autologous flaps.

المهندسة أوعية دموية العضلات رفرف

One of the main factors limiting the thickness of a tissue construct and its consequential viability and applicability in vivo, is the control of oxygen ...

Nanyang Technological University

Research

JoVE Journal

Bioengineering

8.2K Views.  Kaplan Medical Center. To date, thick tissue defects are typically reconstructed by applying autologous tissue flaps or engineered tissues. In this protocol, we present a new method for engineering vascularized tissue flap bearing an autologous pedicle, to serve as a substitute to autologous flaps. 

Video: Engineered Vascularized Muscle Flap

Generation and Grafting of Tissue-engineered Vessels in a Mouse Model

The construction of vascular conduits is a fundamental strategy for surgical repair of damaged and injured vessels resulting from cardiovascular diseases. The current protocol presents an efficient and reproducible strategy in which functional tissue engineered vessel grafts can be generated using partially induced pluripotent stem cell (PiPSC) from human fibroblasts. We designed a decellularized vessel scaffold bioreactor, which closely mimics the matrix protein structure and blood flow that exists within a native vessel, for seeding of PiPSC-endothelial cells or smooth muscle cells prior to grafting into mice. This approach was demonstrated to be advantageous because immune-deficient mice engrafted with the PiPSC-derived grafts presented with markedly increased survival rate 3 weeks after surgery. This protocol represents a valuable tool for regenerative medicine, tissue engineering and potentially patient-specific cell-therapy in the near future.

Here, we present a protocol to generate tissue engineered vessel grafts that are functional for grafting into mice by double seeding partially induced pluripotent stem cell (PiPSC) - derived smooth muscle cells and PiPSC - derived endothelial cells on a decellularized vessel scaffold bioreactor.

جيل ومسمار سفن الهندسة الأنسجة في نموذج الفأر

The construction of vascular conduits is a fundamental strategy for surgical repair of damaged and injured vessels resulting from cardiovascular diseases. ...

Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a deeper comprehension of the neural system scientists aim to simplistically reconstruct the tissue by assembling it in vitro from basic building blocks using a tissue engineering approach. Our group developed a tissue-engineered silk and collagen-based 3D brain-like model resembling the white and gray matter of the cortex. The model consists of silk porous sponge, which is pre-seeded with rat brain-derived neurons, immersed in soft collagen matrix. Polarized neuronal outgrowth and network formation is observed with separate axonal and cell body localization. This compartmental architecture allows for the unique development of niches mimicking native neural tissue, thus enabling research on neuronal network assembly, axonal guidance, cell-cell and cell-matrix interactions and electrical functions.

Insight into the complex actions of the brain requires advanced research tools. Here we demonstrate a novel silk-collagen-based 3D engineered model of neural tissue resembling brain-like architecture. The model can be used to study neuronal network assembly, axonal guidance, cell-cell interactions and electrical activity.

استنادا الحرير الكولاجين المهندسة 3D نموذج الاستقطاب العصبية الأنسجة

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

هندسة الأنسجة عن طريق الجوهرية الأوعية الدموية في<em&gt; في فيفو</em&gt; غرفة هندسة الأنسجة

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from traumatic injury, disease and various congenital, genetic and acquired conditions are quite common. Tissue engineering and regenerative medicine technologies have enormous potential to provide a therapeutic solution. However, utilization of biologically relevant animal models in combination with longitudinal assessments of pertinent functional measures are critical to the development of improved regenerative therapeutics for treatment of VML-like injuries. In that regard, a commercial muscle lever system can be used to measure length, tension, force and velocity parameters in skeletal muscle. We used this system, in conjunction with a high power, bi-phase stimulator, to measure in vivo force production in response to activation of the anterior crural compartment of the rat hindlimb. We have previously used this equipment to assess the functional impact of VML injury on the tibialis anterior (TA) muscle, as well as the extent of functional recovery following treatment of the injured TA muscle with our tissue engineered muscle repair (TEMR) technology. For such studies, the left foot of an anaesthetized rat is securely anchored to a footplate linked to a servomotor, and the common peroneal nerve is stimulated by two percutaneous needle electrodes to elicit muscle contraction and dorsiflexion of the foot. The peroneal nerve stimulation-induced muscle contraction is measured over a range of stimulation frequencies (1-200 Hz), to ensure an eventual plateau in force production that allows for an accurate determination of peak tetanic force. In addition to evaluation of the extent of VML injury as well as the degree of functional recovery following treatment, this methodology can be easily applied to study diverse aspects of muscle physiology and pathophysiology. Such an approach should assist with the more rational development of improved therapeutics for muscle repair and regeneration.

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

We describe an in vivo protocol to measure dorsiflexion of the foot following stimulation of the peroneal nerve and contraction of the anterior crural compartment of the rat hindlimb. Such measurements are an indispensable translational tool for evaluating skeletal muscle pathology and tissue engineering approaches to muscle repair and regeneration.

تطبيقات<em&gt; في فيفو</em&gt; التجارب الفنية من الفئران قصبة الساق الأمامية لتقييم الأنسجة المهندسة وإصلاح العضلات والهيكل العظمي

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from ...

A Model of Free Tissue Transfer: The Rat Epigastric Free Flap

Free tissue transfer has been increasingly used in clinical practice since the 1970s, allowing reconstruction of complex and otherwise untreatable defects resulting from tumor extirpation, trauma, infections, malformations or burns. Free flaps are particularly useful for reconstructing highly complex anatomical regions, like those of the head and neck, the hand, the foot and the perineum. Moreover, basic and translational research in the area of free tissue transfer is of great clinical potential. Notwithstanding, surgical trainees and researchers are frequently deterred from using microsurgical models of tissue transfer, due to lack of information regarding the technical aspects involved in the operative procedures. The aim of this paper is to present the steps required to transfer a fasciocutaneous epigastric free flap to the neck in the rat.
This flap is based on the superficial epigastric artery and vein, which originates from and drain into the femoral artery and vein, respectively. On average the caliber of the superficial epigastric vein is 0.6 to 0.8 mm, contrasting with the 0.3 to 0.5 mm of the superficial epigastric artery. Histologically, the flap is a composite block of tissues, containing skin (epidermis and dermis), a layer of fat tissue (panniculus adiposus), a layer of striated muscle (panniculus carnosus), and a layer of loose areolar tissue.
Succinctly, the epigastric flap is raised on its pedicle vessels that are then anastomosed to the external jugular vein and to the carotid artery on the ventral surface of the rat's neck. According to our experience, this model guarantees the complete survival of approximately 70 to 80% of epigastric flaps transferred to the neck region. The flap can be evaluated whenever needed by visual inspection. Hence, the authors believe this is a good experimental model for microsurgical research and training.

This paper describes the steps required to raise a fasciocutaneous epigastric free flap and transfer it to the neck in the rat.

نموذج من حر نقل الأنسجة: الجرذ شرسوفي الحرة رفرف

Free tissue transfer has been increasingly used in clinical practice since the 1970s, allowing reconstruction of complex and otherwise untreatable defects ...

Medicine

A Reliable Porcine Fascio-Cutaneous Flap Model for Vascularized Composite Allografts Bioengineering Studies

Vascularized Composite Allografts (VCA) such as hand, face, or penile transplant represents the cutting-edge treatment for devastating skin defects, failed by the first steps of the reconstructive ladder. Despite promising aesthetic and functional outcomes, the main limiting factor remains the need for a drastically applied lifelong immunosuppression and its well-known medical risks, preventing broader indications. Therefore, lifting the immune barrier in VCA is essential to tip the ethical scale and improve patients&#39; quality of life using the most advanced surgical techniques. De novo creation of a patient-specific graft is the upcoming breakthrough in reconstructive transplantation. Using tissue engineering techniques, VCAs can be freed of donor cells and customized for the recipient through perfusion-decellularization-recellularization. To develop these new technologies, a large-scale animal VCA model is necessary. Hence, swine fascio-cutaneous flaps, composed of skin, fat, fascia, and vessels, represent an ideal model for preliminary studies in VCA. Nevertheless, most VCA models described in the literature include muscle and bone. This work reports&#160;a reliable and reproducible technique for saphenous fascio-cutaneous flap harvest in swine, a practical tool for various research fields, especially vascularized composite tissue engineering.

The present protocol describes the porcine fascio-cutaneous flap model and its potential use in vascularized composite tissue research.

نموذج رفرف لفافة جلدية خزفية موثوقة لدراسات الهندسة الحيوية المركبة الوعائية

Vascularized Composite Allografts (VCA) such as hand, face, or penile transplant represents the cutting-edge treatment for devastating skin defects, ...

Fabrication of Engineered Vascular Flaps Using 3D Printing Technologies

Engineering implantable, functional, thick tissues requires designing a hierarchical vascular network. 3D bioprinting is a technology used to create tissues by adding layer upon layer of printable biomaterials, termed bioinks, and cells in an orderly and automatic manner, which allows for creating highly intricate structures that traditional tissue engineering techniques cannot achieve. Thus, 3D bioprinting is an appealing in vitro approach to mimic the native vasculature complex structure, ranging from millimetric vessels to microvascular networks. 
Advances in 3D bioprinting in granular hydrogels enabled the high-resolution extrusion of low-viscosity extracellular matrix-based bioinks. This work presents a combined 3D bioprinting and sacrificial mold-based 3D printing approach for fabricating engineered vascularized tissue flaps. 3D bioprinting of endothelial and support cells using recombinant collagen-methacrylate bioink within a gelatin support bath is utilized for the fabrication of a self-assembled capillary network. This printed microvasculature is assembled around a mesoscale vessel-like porous scaffold, fabricated using a sacrificial 3D printed mold, and is seeded with endothelial cells. 
This assembly induces the endothelium of the mesoscale vessel to anastomose with the surrounding capillary network, establishing a hierarchical vascular network within an engineered tissue flap. The engineered flap is then directly implanted by surgical anastomosis to a rat femoral artery using a cuff technique. The described methods can be expanded for the fabrication of various vascularized tissue flaps for use in reconstruction surgery and vascularization studies.

Engineered flaps require an incorporated functional vascular network. In this protocol, we present a method of fabricating a 3D printed tissue flap containing a hierarchical vascular network and its direct microsurgical anastomoses to rat femoral artery.

تصنيع اللوحات الوعائية الهندسية باستخدام تقنيات الطباعة 3D

Engineering implantable, functional, thick tissues requires designing a hierarchical vascular network. 3D bioprinting is a technology used to create ...

Procurement and Perfusion-Decellularization of Porcine Vascularized Flaps in a Customized Perfusion Bioreactor

Large volume soft tissue defects lead to functional deficits and can greatly impact the patient&#39;s quality of life. Although surgical reconstruction can be performed using autologous free flap transfer or vascularized composite allotransplantation (VCA), such methods also have disadvantages. Issues such as donor site morbidity and tissue availability limit autologous free flap transfer, while immunosuppression is a significant limitation of VCA. Engineered tissues in reconstructive surgery using decellularization/recellularization methods represent a possible solution. Decellularized tissues are generated using methods that remove native cellular material while preserving the underlying extracellular matrix (ECM) microarchitecture. These acellular scaffolds can then be subsequently recellularized with recipient-specific cells.
This protocol details the procurement and decellularization methods used to achieve acellular scaffolds in a pig model. In addition, it also provides a description of the perfusion bioreactor design and setup. The flaps include the porcine omentum, tensor fascia lata, and the radial forearm. Decellularization is performed via ex vivo perfusion of low concentration sodium dodecyl sulfate (SDS) detergent followed by DNase enzyme treatment and peracetic acid sterilization in a customized perfusion bioreactor.
Successful tissue decellularization is characterized by a white-opaque appearance of flaps macroscopically. Acellular flaps show the absence of nuclei on histological staining and a significant reduction in DNA content. This protocol can be used efficiently to generate decellularized soft tissue scaffolds with preserved ECM and vascular microarchitecture. Such scaffolds can be used in subsequent recellularization studies and have the potential for clinical translation in reconstructive surgery.

The protocol describes the surgical procurement and subsequent decellularization of vascularized porcine flaps by the perfusion of sodium dodecyl sulfate detergent through the flap vasculature in a customized perfusion bioreactor.

شراء ونزع الخلايا من اللوحات الوعائية الخنازير في مفاعل حيوي مخصص للتروية

Large volume soft tissue defects lead to functional deficits and can greatly impact the patient&#39;s quality of life. Although surgical reconstruction ...

Porcine As a Training Module for Head and Neck Microvascular Reconstruction

Live models that resemble surgical conditions of humans are needed for training free-flap harvesting and anastomosis. Animal models for training purposes have been available for years in many surgical fields. We used the female (because they are easy to handle for the procedure) Yorkshire pigs for the head and neck reconstruction by harvesting the deep inferior epigastric artery perforator or the superior epigastric artery perforator flap. The anastomosis site (neck skin defect or tracheal wall defect) was prepared via the dissection of the common carotid artery and the internal jugular vein, in which 3.5&#215; loupe magnification was used for anastomosis as we use on human cases in real life. This procedure demonstrates a new training method using a reliable learning model and provides a detailed anatomy in a live scenario. We focused on the ischemia time, harvesting, vessel anastomosis, and designing the flap to fit the defect site. This model improves tissue handling and with the use of proper instruments can be repeated many times so that the surgeon is fully confident before starting the surgery on humans.

Here we present a protocol for the use of the pig superior epigastric artery perforator flap as a learning module for head and neck microvascular reconstruction.

الخنزير كنموذج تدريبي للتعمير ميكروفاسكولار الرأس والعنق

Live models that resemble surgical conditions of humans are needed for training free-flap harvesting and anastomosis. Animal models for training purposes ...

Modified Heterotopic Hindlimb Osteomyocutaneous Flap Model in the Rat for Translational Vascularized Composite Allotransplantation Research

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand and face transplants and, more recently, abdominal wall, uterus, and urogenital transplants. Functional outcomes have exceeded initial expectations, and most recipients enjoy an improved quality of life. However, as clinical experience accumulates, chronic rejection and complications from the immunosuppression must be addressed. In many cases where grafts have failed, the causative pathology has been ischemic vasculopathy. <span style="font-size: 11pt; line-height: 115%; font-family: Arial, sans-serif; color: rgb(41, 43, 49); background-image: initial; background-position: initial; background-size: initial; background-repeat: initial; background-attachment: initial; background-origin: initial; background-clip: initial;">The biological mechanisms of the acute and chronic rejection associated with VCA, especially ischemic vasculopathy, are important areas of research. However, due to the very small number of VCA patients, the evaluation of proposed mechanisms is better addressed in an experimental model. Multiple groups have used animal models to address some of the relevant unsolved questions in VCA rejection and vasculopathy. Several model designs involving a variety of species are described in the literature. Here we present a reproducible model of VCA heterotopic hindlimb osteomyocutaneous flap in the rat that can be utilized for translational VCA research. This model allows for the serial evaluation of the graft, including biopsies and different imaging modalities, while maintaining a low level of morbidity.

Vascularized composite allograft offers life-altering benefits to transplant recipients, but the biological causes of graft rejection and vasculopathy remain poorly understood. The rodent surgical model presented here offers a reproducible, clinically relevant model of transplantation, allowing researchers to evaluate rejection events and potential therapeutic strategies to prevent their occurrence.

المعدلة المغايرة الأوستيوميوكوتانيوس رفرف النموذج في الفئران لنقل الاوعيه الدموية المركبة المركب البحوث

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand ...

المهندسة أوعية دموية العضلات رفرف

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