Yazarlar Valerie Schroeder Bu eser teknik yönleri ile ona yardım için teşekkür etmek istiyorum. Geliştirilmesi bu model ile ilgili çalışma Cook biyoteknoloji, Inc. (West Lafayette, inç USA) destek verdi.

Bu iletişim kuralı hayvanlarda kullanımı Üniversitesi Notre Dame kurumsal hayvan bakım ve kullanmak Komitesi tarafından kabul edildi, tüm yasal gereksinimleri ve yönergeleri takip ve tarafından akredite bir tesiste yapılmıştır Değerlendirme ve laboratuvar hayvan bakımı (AAALAC), uluslararası akreditasyon Birliği.
 1. hayvan seçim <ol><li>edinme patojen-Alerjik Sprague Dawley fareler bir saygın satıcı veya damızlık koloni. 
	Not: Çoğu zaman, tesis veteriner veya Yöneticisi uygun kaynakları için araştırmacı yönlendirebilirsiniz. Fareler yaklaşık 250 g vücut ağırlığı bu yordam için uygun.</li>
	<li>Bir gün önce cerrahi, klinik olarak sağlıklı olduklarından emin olmak için kullanılmak üzere hayvanlar inceleyin. Bu bağlamda, hayvan tartmak ve vücut durumunu değerlendirmek; solunum, herhangi bir açık yarası veya kitleler varlığı ve hayvan genel etkinlik düzeyini herhangi bir anormallik unutmayın. Bu tür gözlemler potansiyel dışlama ölçütü olarak düşünün ve cerrahi kayıttaki herhangi bir anormallik Not.</li>
</ol> 2. Anestezi ve aseptik cerrahi için hayvan hazırlık <ol><li>analjezi, 1.1-1.2 mg/kg sürekli yayın buprenorfin, subkutan sıçan önce ameliyat. Uygula oftalmik merhem için her iki gözde 2 h için yönetmek ve yönetmek 2 mL steril serum fizyolojik subkutan ısındı.</li>
	<li>Fareyi ketamin hidroklorür (90 mg/kg) ve xylazine (10 mg/kg) mayi bir doz ile anestezi. Oküler yağ küçük bir miktar her gözün kornea dessication önlemek için geçerlidir. Bir kez belgili tanımlık hayvan &#8217; s solunum sabit ve artık bir firma ayak çimdik için herhangi bir yanıt olduğunu, fareyi anestezi ameliyat için yeterli bir seviyeye ulaşmıştır. Anestezi kabul edilebilir bir düzeyde daha fazla derin, varlığı ile teyit edilmesi normal solunum ve normal renk göz (yetersiz kardiyovasküler perfüzyon öneren karşı solgunluk,).</li>
	<li>Ventral karın ile göğüs kemiği için apış arasından ve lateral kenar boşluklarına ventrum üzerinde bir ticari clippers tıraş. Nemli kağıt havlu bölgeye uygulayarak kısaltıldı tüylerden.</li>
	<li>% 70 etanol ile silin ve ardından traş alan bir iodophor antiseptik ile fırçalayın. Belgili tanımlık kesme için bir pencere potansiyel kirlenme ayrılmış olabilir cerrahi perdeler uygulamak.</li>
	<li>Steril Cerrahi önlük, bir yüz maskesi ve saç Bone don. İyice yıkayın ve eller sabun ve don steril cerrahi eldivenler ile fırçalayın.</li>
</ol> 3. Oluşturma ve Ventral karın fıtığı tamiri <ol><li>ameliyat öncesi cerrahi aletler otoklav tarafından sterilize.</li>
	<li>Bir neşter kullanarak, yaptığınız bir karın kas düzeyine ventral orta çizgi boyunca yaklaşık 3-4 cm dikey kesi. Karın duvarının yavaşça bir Forseps ile Asansör ve linea küçük bir açıklıktan oluşturmak alba. Temel iç organlar korumak için bir forseps delikten ve kas duvar oluşturmak için kazıma gibi koruyucu yol gösterici için karın boşluğu içine ekleyin bir yaklaşık 1 cm lateral her iki yanına kesme yaklaşık 2 cm x 2 cm tam kalınlıkta kusur, orta çizgi üzerinde her iki ucunda da linea, alba kesi.</li>
	<li>Lay önceden kesilmiş 2 cm x 2 cm Bölüm 4 katmanlı SIS (şekil 1), bu durumda mekanik köprüleme malzemenin kusur. 
	Not: Alternatif olarak, bir altlık yaklaşım malzeme bölümünü yerleştirdiği karın boşluğuna kusur iç boyutu gelen karşılamak için kullanılabilir.</li>
	<li>Dikiş köprüleme, dört köşe 4-0 ipek ya da naylon bir basit kesintiye uğramış dikiş deseni kullanarak karın duvarı kas malzeme. 
	Not: Absorbe olmayan dikiş kolay tanımlanması hasat absorbe suture maddi karşı üzerine yerleştirilmiş malzemenin kullanılmasına.</li>
	<li>Malzeme her kenarı eşit aralıklı üç ek dikiş ile güvenli. İç organlar karın duvarından geçişini sağlayan önemli boşluklar yok emin olmak için tüm çevresel kenarına incelemek; ve böyle boşluklar varsa, kenar boşluğunu güvenliğini daha da artırmak için ek dikiş yerleştirin.</li>
	<li>4-0 absorbe dikiş basit kesintiye uğramış desen ile subkutan doku kapatın. Basit bir kesintiye uğramış desen uygulanan dikiş de kullanılabilir olsa cilt cerrahi zımba ile yakın.</li>
	<li>Bırakmayın hayvan katılımsız sternal recumbency; yeniden elde etmek için yeterli kendine geldi kadar ve tam olarak anesteziden kurtarıldı kadar diğer hayvanların şirkete cerrahi uğramıştır bir hayvan döndürmüyor. Bu süre boyunca, hayvan hipotermi önlemek için sıcak bir alanda devam.</li>
	Tam kurtarma ve normal iyileşme sağlamak için bu yordamı uğramıştır <li>inceleyin hayvan her gün. Deri zımba zımba sökücü ile 7-10 gün ameliyatı takiben kaldırmak.</li>
</ol> 4. Hasat implante malzeme <ol><li>tutarlı çalışma ihtiyaçları, ötenazi, yönetilen bir odasında karbon dioksit aşırı doz ile 28 gün ameliyatı takiben 10 ila 30 oranında, önceden belirlenmiş bir zamanda, genellikle 21 - hayvanlar doldurun.</li>
	<li>Kullanarak standart nekropsi araçlar, açık belgili tanımlık deri ve karın duvarı ve implantın alan bulaşmasına neden. 
	Not: Eğer absorbe olmayan dikiş implant güvenliğini sağlamak için kullanılan malzeme orijinal dikiş (Şekil 2) tarafından belirlenebilir.</li>
	<li>Herhangi bir olumsuz tepkiler (örneğin, yara yırtılması, apse, seroma, Hematom) ve yapışma ölçüde ve azim temel iç organlar için de dahil olmak üzere brüt herhangi bir patolojik gözlem gözlemlemek.</li>
	<li>İmplant bölgesi doku implant içeren geniş bir bölümünü keserek çıkarın. Örnek işleme histolojik inceleme ve karakterizasyonu için hazırlanırken % 10 tarafsız tampon formalin gibi bir sabitleştirici yerleştirin.</li>
</ol>

<ol>
	<li>Poussier, 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=A+review+of+available+prosthetic+material+for+abdominal+wall+repair.">A review of available prosthetic material for abdominal wall repair.</a> J. Visc. Surg. 150 (1), 52-59 (2013).</li><li>Hjort, H., Mathisen, T., Alves, A., Clermont, G., Boutrand, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-year+results+from+a+preclinical+implantation+study+of+a+long-term+resorbable+surgical+mesh+with+time-dependent+mechanical+characteristics.">Three-year results from a preclinical implantation study of a long-term resorbable surgical mesh with time-dependent mechanical characteristics.</a> Hernia. 16 (2), 191-197 (2012).</li><li>Ko, R., Kazacos, E. A., Snyder, S., Ernst, D. M. J., Lantz, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tensile+strength+comparison+of+small+intestinal+submucosa+body+wall+repair.">Tensile strength comparison of small intestinal submucosa body wall repair.</a> J. Surg. Res. 135 (1), 9-17 (2006).</li><li>Garcia-Moreno, F., 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=Comparing+the+host+tissue+response+and+peritoneal+behavior+of+composite+meshes+used+for+ventral+hernia+repair.">Comparing the host tissue response and peritoneal behavior of composite meshes used for ventral hernia repair.</a> J. Surg. Res. 193 (1), 470-482 (2015).</li><li>Soiderer, E. E., Lantz, G. C., Kazacos, E. A., Hodde, J. P., Wiegand, R. 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Y., Wang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biomechanical+behavior+and+host+response+to+porcine-derived+small+intestine+submucosa,+pericardium+and+dermal+matrix+acellular+grafts+in+a+rat+abdominal+defect+model.">The biomechanical behavior and host response to porcine-derived small intestine submucosa, pericardium and dermal matrix acellular grafts in a rat abdominal defect model.</a> Biomaterials. 32 (29), 7086-7095 (2011).</li><li>Ma, J., Sahoo, S., Baker, A. R., Derwin, K. 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Bir yazar (CJ) Cook biyoteknoloji, Inc, Medikal dereceli SIS üreticisine istihdam edilmektedir.

Malzemelerin ventral karın fıtık onarımı için büyük ilgi, özellikle de bir ilk mekanik köprü sağlamak ve sonra ana bilgisayar doku dahil edebiliyoruz. Bu bağlamda, malzemelerin çeşitli araştırdık, SIS, domuz acellular dermal matris ve domuz kalp zarını gibi. 6 , 7 bu malzemelerin bu dokuların ekstraselüler matrisleri temsil ve böylece doku birleşme kolaylaştırmak içine hücre göç ve çoğalırlar, iskele hareket.
<p class="jove_co...

Karın duvarı fıtığı Konjenital defekt, travma olabilir veya cerrahi yaralar karın duvarı içeren başarısız kapatılması sonucunda oluşabilecek bir sık karşılaşılan klinik bir konudur. Onarım yaygın bir implant indirimli bu kime karın duvarı kapalıdır sadece dikiş ile karşılaştırıldığında bir implant ile tedavi edilen hastaların kaydetti Rekürrens ile karın duvarının güçlendirmek kullanmayı gerektirir. 1
Cerrahi fıtık onarımı kez kusur yerleştirilmiş bir malzeme ile mekanik köprü oluşturma gerektirir. Bu bağlamda, sentetik ve doğal malzemeler fıtık onarımı için kullanılmaktadır. Birçok sentetik malzeme genellikle sigara absorbe ve diğer sentetik implantlar bir absorbe olmayan malzemelerin bir absorbe ile birleştirmek polipropilen, polietilen tereftalat polyester ve genişletilmiş politetrafloroetilen, yer Polimer polygalactine gibi. 1 buna ek olarak, doğal malzeme implantlar genellikle hayvan veya insan Kadavra kaynaklardan elde. Fıtık onarımı için kullanılan doğal malzemeler genellikle kollajen zengindir ve ingrowth ve ana bilgisayar doku bütünleştirilmesi için bir iskele olarak hareket.
Arzu geliştirmek ve yeni malzemeler ventral karın fıtık onarımı için en iyi duruma getirmek için aday malzemelerin değerlendirilmesi için uygun bir hayvan modeli gerektirir. Türler, koyun,2 domuz,3 tavşan,4 fareler,5,6,7,8 ve köpekler de dahil olmak üzere çeşitli çalışmalar yapılmıştır. 9 kolayca işlenmiş bir modeli doğuştan suşların genetik değişkenliği denetimi için izin çeşitli mevcuttur sundukları fareler gibi çalışmalar için kullanmanın avantajı var. Buna ek olarak, daha küçük genel vücut yüzey alanına yukarıda listelenen diğer türler ile karşılaştırıldığında sıçan izin doğada deneysel olabilir malzemeleri değerlendirilmesi ve bu nedenle, değil bol.  Rat karşı diğer türler ile ilgili daha fazla, azaltılmış maliyetleri nispeten yüksek ekran aday malzemeler kullanmak ventral karın fıtığı tamiri için yeteneği sağlar.  Malzemeler için konuları çoğunlukla cerrah, gücü, yeteneği ana bilgisayar doku, Biyouyumluluk ve enfeksiyona direnç içine dahil tarafından işleme kolaylığı içerir.
Örnek olarak, domuz küçük bağırsak submukoza (SIS) doku onarım endikasyonları, ventral karın, kasık ve diyafragmatik fıtığı tamiri dahil olmak üzere çok çeşitli için kullanılan doğal bir biomaterial var. 10 , 11 , 12 kardeşim hızla ana bilgisayar doku kurulmuştur ve enfeksiyon genellikle dayanıklı olduğu tespit edilmiştir ve çok malzeme kirlenmiş alanlarda belirli yarar vardır. 13 , 14 sıçan modelinde, işleme, iyi gerilme kuvveti uniaxial çekme hatası test ve doku ingrowth kollajen birikimi ve neovaskülarizasyon açısından teşviki tarafından belirlenen kolaylığı SIS göstermiştir. 6 , 7
Doğru cerrahi kusur köprüleme bir temel üretken modelleme aday tamir harçları değerlendirirken özelliğidir. Hayvan yeterince imzalat ve aseptik teknik kesinlikle takip olması gerekir. Ayrıca, standart boyutta bir karın duvarı defekti dikkatli oluşturulmalı ve yeterince malzeme defekt kenar boşluklarına güvenlik altına alır bir şekilde köprü. 8 bu iletişim kuralı, tamiri ve cerrahi oluşturma ventral karın fıtığı SIS kullanarak bir sıçan malzeme köprü oluşturma bir örnek olarak takip edilmesi gereken standart bir yöntemini sağlar.

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Isoflurane (Isoflo)</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">17579</td>
			<td style="width:167px;">gas anesthetic</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Optixcare Eye Lubricant</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td style="width:119px;">OPX4240</td>
			<td>Ocular lubricant</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Oster clippers</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">6092</td>
			<td>hair clippers</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Betadine surgical scrub</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">1618</td>
			<td>antiseptic iodophor</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Sterile scalpel</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">329</td>
			<td>#10 scalpel</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">9&#34; x 12&#34; disposable surgical drape</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">SP-RPS</td>
			<td>Surgical drape</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">4-0 nylon suture</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">SUT 812</td>
			<td>Suture material</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">4-0 Absorbable Suture</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">29242</td>
			<td>Synthetic absorbable suture</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">9 mm Autoclips and applier</td>
			<td style="width:71px;">Braintree Scientific, Inc.</td>
			<td style="width:119px;">ACS KIT</td>
			<td>Surgical staples for closure of skin incision</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Torbugesic/analgesic</td>
			<td style="width:71px;">Henry Schine Animal Health</td>
			<td align="right" style="width:119px;">12084</td>
			<td>Butorphanol tartarate for post-operative analgesia</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Small intestinal submucosa (SIS)</td>
			<td style="width:71px;">Cook Biotech, Inc.</td>
			<td style="width:119px;"></td>
			<td>Implant material for bridging experimental abdominal wall defect</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Rats</td>
			<td style="width:71px;">Harlan, Inc.</td>
			<td style="width:119px;">Sprague Dawley</td>
			<td>Animal for hernia modeling</td>
		</tr>
	</tbody>
</table>

use of a rat model to study ventral abdominal hernia repair

Ventral karın fıtığı bazen herniorraphy (cerrahi onarım) gerektirir nispeten yaygın bir klinik durumdur. Ventral karın fıtığı tamiri genellikle karın duvarı üründe arasında mekanik bir köprü hizmet için bir malzemenin implantasyonu gerektirir. Biyomalzeme, domuz küçük bağırsak submukoza (SIS), gibi Ayrıca hücre büyüme içine implant için bir kafes olarak hizmet ve doğal olarak ana bilgisayar doku dahil edebilirsiniz. Bu tür tamir harçları gelişimi bir tekrarlanabilir biçimde onarmak için deneysel karın duvarı defektleri kolayca oluşturulur ve mükellef bulunmaktadır hayvan modelleri kullanımını yararlanır. Burada sunulan yöntem bir sıçan modelinde ventral karın fıtığı tamiri ve cerrahi oluşturma açıklar. Kardeşim bu modelde deneysel bir ventral karın fıtığı onarmak için kullanıldığında, bu hızla implantasyon 28 gün içinde ana bilgisayar doku kurulmuştur. Histolojik olarak, implante maddi ortaklığın ana bilgisayar doku içine sağlam bir fibrovascular yanıt tarafından karakterizedir. Büyük olasılıkla gelecekteki iyileştirmeler ve uygulamaları sıçan karın fıtığı modelinin daha yakından kopyalama ortak komorbiditeler bir adam için bir araç olarak diyabetik ve/veya obez hayvan içerebilir.

İmplantasyonu sonrası 28 gün tarafından SIS genellikle ana bilgisayar doku (Şekil 2) içine iyi birleşme gösterir. Çoğu durumda, orada 's kalan SIS belirgin, doku kez implante malzeme tanımlamak için keşfedilmeyi gerekir. Histolojik analizler iyi doku birleşme öncelikle fibrovascular doku ve kaydetti çok az kalıntı SIS (şekil 3) ile gösteren brüt patolojik gözlemlere doğruladı. Bu sonuçlar yardımcı programı sıçan ventral karı...

Watch this Scientific Journal Video about Use of a Rat Model to Study Ventral Abdominal Hernia Repair at JoVE.com

Use of a Rat Model to Study Ventral Abdominal Hernia Repair

Bu el yazması oluşturma ve ventral karın duvarı defekti (fıtık) tamiri ile ilgili yöntemler açıklanır. Bu model onarım stratejileri implante malzemeleri kullananlar gibi çalışma için kullanılabilir. Bu makale, domuz küçük bağırsak submukoza ile deneysel fıtık tamiri örnek olarak sunulur.

Ventral karın fıtık onarımı çalışmaya bir fare modeli kullanımı

Ventral abdominal hernia is a relatively common clinical condition that sometimes requires herniorraphy (surgical repair). The repair of ventral abdominal ...

use-of-a-rat-model-to-study-ventral-abdominal-hernia-repair

Research

JoVE Journal

Bioengineering

9.1K Görüntüleme.  University of Minnesota. Bu el yazması oluşturma ve ventral karın duvarı defekti (fıtık) tamiri ile ilgili yöntemler açıklanır. Bu model onarım stratejileri implante malzemeleri kullananlar gibi çalışma için kullanılabilir. Bu makale, domuz küçük bağırsak submukoza ile deneysel fıtık tamiri örnek olarak sunulur.

Video: Use of a Rat Model to Study Ventral Abdominal Hernia Repair

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells behave differently when they are grown within a 3D extra-cellular matrix and also interact with other cells (1-5). Mouse models have been broadly utilized to study tissue morphogenesis in vivo. However mouse and human skin have significant differences in cellular architecture and physiology, which makes it difficult to extrapolate mouse studies to humans. Since melanocytes in mouse skin are mostly localized in hair follicles, they have distinct biological properties from those of humans, which locate primarily at the basal layer of the epidermis. The recent development of 3D human skin reconstruct models has enabled the field to investigate cell-matrix and cell-cell interactions between different cell types. The reconstructs consist of a "dermis" with fibroblasts embedded in a collagen I matrix, an "epidermis", which is comprised of stratified, differentiated keratinocytes and a functional basement membrane, which separates epidermis from dermis. Collagen provides scaffolding, nutrient delivery, and potential for cell-to-cell interaction. The 3D skin models incorporating melanocytic cells recapitulate natural features of melanocyte homeostasis and melanoma progression in human skin. As in vivo, melanocytes in reconstructed skin are localized at the basement membrane interspersed with basal layer keratinocytes. Melanoma cells exhibit the same characteristics reflecting the original tumor stage (RGP, VGP and metastatic melanoma cells) in vivo. Recently, dermal stem cells have been identified in the human dermis (6). These multi-potent stem cells can migrate to the epidermis and differentiate to melanocytes.

In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.

Üç Boyutlu İnsan Cilt Model Yeniden İnşa: Aracı Normal Cilt ve Melanom İlerleme Eğitim

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

Biyomühendislik

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic valve leaflets are composed of interstitial cells suspended within an extracellular matrix (ECM) and are lined with an endothelial cell monolayer. The valve withstands a harsh, dynamic environment and is constantly exposed to shear, flexion, tension, and compression. Research has shown calcific lesions in diseased valves occur in areas of high mechanical stress as a result of endothelial disruption or interstitial matrix damage1-3. Hence, it is not surprising that epidemiological studies have shown high blood pressure to be a leading risk factor in the onset of aortic valve disease4. 
The only treatment option currently available for valve disease is surgical replacement of the diseased valve with a bioprosthetic or mechanical valve5. Improved understanding of valve biology in response to physical stresses would help elucidate the mechanisms of valve pathogenesis. In turn, this could help in the development of non-invasive therapies such as pharmaceutical intervention or prevention. Several bioreactors have been previously developed to study the mechanobiology of native or engineered heart valves6-9. Pulsatile bioreactors have also been developed to study a range of tissues including cartilage10, bone11 and bladder12. The aim of this work was to develop a cyclic pressure system that could be used to elucidate the biological response of aortic valve leaflets to increased pressure loads. 
The system consisted of an acrylic chamber in which to place samples and produce cyclic pressure, viton diaphragm solenoid valves to control the timing of the pressure cycle, and a computer to control electrical devices. The pressure was monitored using a pressure transducer, and the signal was conditioned using a load cell conditioner. A LabVIEW program regulated the pressure using an analog device to pump compressed air into the system at the appropriate rate. The system mimicked the dynamic transvalvular pressure levels associated with the aortic valve; a saw tooth wave produced a gradual increase in pressure, typical of the transvalvular pressure gradient that is present across the valve during diastole, followed by a sharp pressure drop depicting valve opening in systole. The LabVIEW program allowed users to control the magnitude and frequency of cyclic pressure. The system was able to subject tissue samples to physiological and pathological pressure conditions. This device can be used to increase our understanding of how heart valves respond to changes in the local mechanical environment.

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

A cyclic pressure bioreactor capable of subjecting heart valve tissue to physiological and pathological pressure conditions has been designed. A LabVIEW program allows users to control pressure magnitude, amplitude and frequency. This device can be used to study the mechanobiology of heart valve tissue or isolated cells.

Devirli Basınç Biyoreaktör Tasarım Ex vivo</emAort Kalp Vanalar> Eğitim

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges1,2. These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production3. In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (&lambda;=532 nm, Fluence~110 J/cm2, spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 &plusmn; 6 kPa, (n=30). The adhesion strength drops to 2 &plusmn; 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

Fabrikasyon ve Lazer Doku Onarımı Rose Bengal-kitosan Film Uygulamaları

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two ...

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This technique allows for easier venous cuff-anatomoses after a reasonable learning curve. The device is composed of two handles, which are carved out from scalpel blades, one approximator, which is obtained by modifying Kocher's forceps, and cuffs designed from fine-bore polyethylene tubing. The whole process can be performed at a low-cost using common laboratory material. The present report provides a step-by-step protocol for the design of the required pieces and includes stencils.

We describe the design of the &#x201C;quick-linker&#x201D; device for easier orthotopic rat liver transplantation.

Kolay Sıçan Karaciğer Nakli için üretim cihazlar ve Araçlar

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This ...

Athymic Rat Model for Evaluation of Engineered Anterior Cruciate Ligament Grafts

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without intervention. Current treatment strategies include ligament reconstruction with either autograft or allograft, which each have their associated limitations. Thus, there is interest in designing a tissue-engineered graft for use in ACL reconstruction. We describe the fabrication of an electrospun polymer graft for use in ACL tissue engineering. This polycaprolactone graft is biocompatible, biodegradable, porous, and is comprised of aligned fibers. Because an animal model is necessary to evaluate such a graft, this paper describes an intra-articular athymic rat model of ACL reconstruction that can be used to evaluate engineered grafts, including those seeded with xenogeneic cells. Representative histology and biomechanical testing results at 16 weeks postoperatively are presented, with grafts tested immediately post-implantation and contralateral native ACLs serving as controls. The present study provides a reproducible animal model with which to evaluate tissue engineered ACL grafts, and demonstrates the potential of a regenerative medicine approach to treatment of ACL rupture.

Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.

Engineered Ön Çapraz Bağ Greftlerinin Değerlendirme Atimik Rat Modeli

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without ...

In Vitro Permeation of FITC-loaded Ferritins Across a Rat Blood-brain Barrier: a Model to Study the Delivery of Nanoformulated Molecules

Brain microvascular endothelial cells, supported by pericytes and astrocytes endfeet, are responsible for the low permeation of large hydrosoluble drugs through the blood-brain barrier (BBB), causing difficulties for effective pharmacological therapies. In recent years, different strategies for promoting brain targeting have aimed to improve drug delivery and activity at this site, including innovative nanosystems for drug delivery across the BBB. In this context, an in vitro approach based on a simplified cellular model of the BBB provides a useful tool to investigate the effect of nanoformulations on the trans-BBB permeation of molecules. This study describes the development of a double-layer BBB, consisting of co-cultured commercially available primary rat brain microvascular endothelial cells and astrocytes. A multiparametric approach for the validation of the model, based on the measurement of the transendothelial electrical resistance and the apparent permeability of a high molecular weight dextran, is also described. As proof of concept for the employment of this BBB model to study the effect of different nanoformulations on the translocation of fluorescent molecules across the barrier, we describe the use of fluorescein isothiocyanate (FITC), loaded into ferritin nanoparticles. The ability of ferritins to improve the trans-BBB permeation of FITC was demonstrated by flux measurements and confocal microscopy analyses. The results suggest this is a useful system for validating nanosystems for delivery of drugs across the BBB.

In Vitro Permeation of FITC-loaded Ferritins Across a Rat Blood-brain Barrier: a Model to Study the Delivery of Nanoformulated Molecules

A method to establish an in vitro model of blood-brain barrier based on a co-culture of rat brain microvascular endothelial cells and astrocytes is described and validated. This system proved to be a valid tool to study the effect of nanoformulation on the trans-barrier permeation of fluorescent molecules.

Bir Sıçan Kan beyin bariyeri karşısında FITC yüklü ferritinin İn Vitro Geçirgenlik: Bir Model Nanoformulated Moleküllerin Teslim Eğitim için

Brain microvascular endothelial cells, supported by pericytes and astrocytes endfeet, are responsible for the low permeation of large hydrosoluble drugs ...

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.

Uygulamaları<em&gt; İn Vivo</em&gt; Doku değerlendirilmesi için Rat Tibialis Anterior Fonksiyonel Test İskelet Kası Onarım Engineered

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

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in pathological situations such as cancer, vascularization plays a leading role in disease progression. Consequently, there is a strong need for a standardized and well-characterized in vivo model in order to elucidate the mechanisms of neovascularization and develop different vascularization approaches for tissue engineering and regenerative medicine.
We describe a microsurgical approach for a small animal model for induction of a vascular axis consisting of a vein and artery that are anastomosed to an arteriovenous (AV) loop. The AV loop is transferred to an enclosed implantation chamber to create an isolated microenvironment in vivo, which is connected to the living organism only by means of the vascular axis. Using 3D imaging (MRI, micro-CT) and immunohistology, the growing vasculature can be visualized over time. By implanting different cells, growth factors and matrices, their function in blood vessel network formation can be analyzed without any disturbing influences from the surroundings in a well controllable environment. 
In addition to angiogenesis and antiangiogenesis studies, the AV loop model is also perfectly suited for engineering vascularized tissues. After a certain prevascularization time, the generated tissues can be transplanted into the defect site and microsurgically connected to the local vessels, thereby ensuring immediate blood supply and integration of the engineered tissue. By varying the matrices, cells, growth factors and chamber architecture, it is possible to generate various tissues, which can then be tailored to the individual patient's needs.

The Arteriovenous AV Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

We describe a microsurgical approach for the generation of an arteriovenous (AV) loop as a model for analyzing vascularization in vivo in an isolated and well-characterized environment. This model is not only useful for the investigation of angiogenesis, but is also optimally suited for engineering axially vascularized and transplantable tissues.

Bir Küçük Hayvan Modelinde Arteriovenöz (AV) Döngü Anjiogenez ve Vaskülarize Doku Mühendisliği Eğitim için

A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in ...

The Use of a &#946;-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions

Biosensors are becoming increasingly important and implemented in various fields such as pathogen detection, molecular diagnosis, environmental monitoring, and food safety control. In this context, we used &#946;-lactamases as efficient reporter enzymes in several protein-protein interaction studies. Furthermore, their ability to accept insertions of peptides or structured proteins/domains strongly encourages the use of these enzymes to generate chimeric proteins. In a recent study, we inserted a single-domain antibody fragment into the Bacillus licheniformis BlaP &#946;-lactamase. These small domains, also called nanobodies, are defined as the antigen-binding domains of single chain antibodies from camelids. Like common double chain antibodies, they show high affinities and specificities for their targets. The resulting chimeric protein exhibited a high affinity against its target while retaining the &#946;-lactamase activity. This suggests that the nanobody and &#946;-lactamase moieties remain functional. In the present work, we report a detailed protocol that combines our hybrid &#946;-lactamase system to the biosensor technology. The specific binding of the nanobody to its target can be detected thanks to a conductimetric measurement of the protons released by the catalytic activity of the enzyme.

In this work, we report a new method to study protein-protein interactions using a conductimetric biosensor based on the hybrid &#946;-lactamase technology. This method relies on release of protons upon hydrolysis of &#946;-lactams.

Biyomoleküler etkileşimleri algılamak için bir Conductimetric Biyoalgılayıcı β-lactamase tabanlı tahlil kullanımı

Biosensors are becoming increasingly important and implemented in various fields such as pathogen detection, molecular diagnosis, environmental ...

An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) is a surgical technique leading to bone regeneration after an osteotomy. The osteotomized extremities are moved away from each other by gradual distraction to reach the desired elongation. This procedure is widely used in humans for lower and upper limb lengthening, treatment after a bone nonunion, or the regeneration of a bone defect following surgery for bone tumor excision, as well as in maxillofacial reconstruction. Only a few studies clearly demonstrate the efficiency of their protocol in obtaining a functional regenerated bone, i.e., bone that will support physiological weight-bearing without fracture after removal of the external fixator. Moreover, protocols for DO vary and reproducibility is limited by lack of information, making comparison between studies difficult. The aim of this study was to develop a reproducible protocol comprising an appropriate external fixator design for rat limb lengthening, with a detailed surgical technique that permits physiological weight-bearing by the animal after removal of the external fixator.

This study describes a reproducible and detailed protocol using a newly developed external fixator for distraction osteogenesis (DO) in a femoral rat model which permits physiological weight-bearing by the animal after removal of the external fixator.

Oyalama Osteogenesis fonksiyonel rejenere uyluk için önde gelen bir fare modeli için verimli ve tekrarlanabilir bir protokol

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) ...

Ventral karın fıtık onarımı çalışmaya bir fare modeli kullanımı

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