Bu çalışma kısmen El Pilot Ödülü ve NIH / NIAMS 1K01AR068386-01 (AEL kadar) ve NIAMS / NIH P30AR061307 Cerrahisi Amerikan Derneği tarafından desteklenmiştir.

Rochester Üniversitesi&#39;nde Hayvan Araştırmaları Üniversite Komitesi, tüm hayvan deneyleri onayladı. On-12 haftalık dişi C57BL / 6J fareleri kullanılmıştır. Flexor Tendon Cerrahisi Animals 1. Hazırlık (~ 15 dk) <ol><li> Otoklav cerrahi aletler, sterilize boyunca steril eldiven giyin ve steril bir operasyon alanı korumak için. </li><li> vücut ağırlığına tekabül eden bir ketamin hacmi (80 mg / kg) ve ksilazin (10 mg / kg) ile intraperitoneal enjeksiyon (i.p.) ile fare anestezisi. ayak-tutam refleksinin yokluğu yoluyla sedasyon derinliği onaylayın. </li><li> subkutan enjeksiyon yoluyla rüçhan analjezi (buprenorfin, 0.05-0.1 mg / kg) yönetmek. kurumasını önlemek gözleri oftalmik merhem sürün. </li><li> tüm arka bacak üzerinde kürk kırpma cerrahi bölgeyi hazırlayın. povidon iyot ile tekrar% 70 etanol ve ardından povidon iyot ile sırayla, ovma cildi sterilize ve. </li></ol><ol><li> buzağı medial yüzeysel görülen fleksör digitorum longus (FDL) tendon, bulun. Bir neşter kullanılarak, tendon ortaya çıkarmak için mikro-makas ile deride küçük 0.5-1 cm&#39;lik kesi yapmak. </li><li> çevreleyen dokudan FDL tendon ayırmak ve myotendinous kavşağa o kadar izlemek için forseps kullanabilir. posterior tibial arter önlemek için özen, onu serbest bırakmak için yaylı makas ile bu kavşakta tendon kesin. </li><li> 5-0 naylon sütür ile cilt kapatın. Not: FDL transection ve tamiri (2.4 adımlar) Stereomikroskopta kullanılarak yapılmalıdır. </li><li> bahar mikro-makas kullanılarak arka pençe posterolateral yönü üzerinde 3 mm kesi yapmak. Yavaşça doku hasarı en aza indirmek ve FDL tendon tanımlamak için emin olun, forseps ile çevreleyen yumuşak doku ve kas geri çekin. <ol><li> Yavaşça FDL tendon yükseltmek ve tamamen mil kullanarak transectcro-makas. periyodik serum fizyolojik ile ıslatma yoluyla tendonun kurumasını önlemek, 8-0 sütür ile modifiye Kessler desen tendon birlikte FDL uçlarını dikin. </li><li> tendon üzerinde yumuşak doku ve kas değiştirin sonra 5-0 naylon sütür ile kesi kapatın. </li></ol></li><li> anestezi kurtarıldı kadar vücut ısısını korumak için, 37 ° C slayt sıcak seti fareler yerleştirin. </li><li> ameliyat sonrası artmış vokalli ve karıştırdı kürk dahil olmak üzere yetmezliği ya da enfeksiyon belirtileri için fareler izleyin. Farelerde kadar her 12 saatte artık ağrı veya sıkıntı belirtileri gösterebilir analjezik olarak buprenorfin (50 ul / fare) enjekte edilir. </li><li> fareler sonraki adımlara devam etmeden önce 10 gün sonrası cerrahi bir süre için iyileşmek için izin verin. Not: Hücreler herhangi bir zaman noktası sonrası cerrahi hasat edilebilir. Burada, 10 gün, şu anda doku nispeten yüksek selülarite verilen bu Örnek deneyler için seçilir. </li></ol> 3. LabeliBisiklet Hücreler ng (~ 10 dakika) <ol><li> Edu (5-etinil-2&#39;deoxyuridine) çözeltisi hazırlayın (steril fosfat tamponlu tuzlu su içinde 10 mg / ml [PBS] +,% 1 dimetil Sülfoksit [DMSO] çözünürlüğünü arttırmak için). </li><li> IP enjeksiyon yoluyla kurban önce 24 saat 100 ul edu (50 mg / kg) ile fareler enjekte edilir. </li></ol> Sitoloji Santrifüj 4. Hasat Hücreleri (2.5 saat, ~ 30 dakika Hands-Zaman) <ol><li> PBS içinde 3 mg / ml kollajenaz I hazırlayın. </li><li> en fazla 3 L / dakikalık bir akış oranında karbon dioksit ile boğularak ile fare öldürülür ve ikincil bir tedbir olarak servikal dislokasyon gerçekleştirin. </li><li> Yukarıdaki adım 2.4 kullanılarak onarılan bir tendonun bulun ve mikro makasla yaralanma bölgesinde her iki tarafında yaklaşık 2 mm keserek tendon dokusuna izole eder. <ol><li> 3 mg / ml kolajenaz 1 ml bir petri neşter ile kıyma doku daha sonra dokunun kırmak için 18 G iğneli birkaç kez ile karışım geçmektedir. 1.5 ml&#39;lik bir santrifüj tüpüne karışımın toplanması. </li&gt; <li> çalkalanarak 37 ° C&#39;de 1 saat süre ile tendon dokusuna Digest. </li><li> Sıkma dokusunu (5 dakika için 300 x g), aspirat kolajenaz, daha sonra birkaç kez pipetli yukan aşağı çekilerek PBS içinde 500 ul% 3 sığır serum albümin (BSA) ile hücre / doku tekrar süspansiyon. </li><li> 70 mikron hücre süzgecinden Filtre hücre süspansiyonu enkaz ve tendonun büyük parça ve 37 ° C inkübatör kenara kaldırın. </li><li> Başka bir 1.5 ml santrifüj tüpüne tendon dokusuna yerleştirin ve pipetleme, 1 mi kolajenaz çözeltisi ile yenileme aşağı karıştırın. </li><li> çalkalanarak 37 ° C&#39;de doku bir saat inkübe edin. </li><li> sindirilmiş tendon dokusu (5 dakika boyunca 300 xg) aşağı Spin, sonra yukarı pipetleme ve birkaç kez aşağı PBS içinde 500 ul% 3 BSA içinde tekrar süspansiyon. </li><li> , 70 mikron hücre süzgecinden hücre süspansiyonu Filtre 4.3.3 izole süspansiyon ile birleştirmek, daha sonra bir hemasitometre hücreleri saymak. </li><li> Daha sonra 3-6 kadar tekrar süspansiyon, PBS içinde% 3 BSA ile bir kez daha yıkama hücrelerix 10 4/100 ul. </li></ol></li><li> pozitif yüklü slayt bir sitoloji huni takın ve slayt taşıyıcısının içine kilitleyin. Bir sitoloji santrifüj içine tüm aparat takın ve huni 100 ul hücre süspansiyonu ekleyin. 5 dakika boyunca 300 xg&#39;de santrifüj slayta hücreleri dağıtmak için. </li></ol> 5. İmmünositokimya Edu Algılama için (2 saat, ~ 45 dakika Hands-Zaman) Not: Tüm bekletme işlemleri florofor photobleaching sınırlamak için karanlıkta yapılmalıdır. <ol><li> adımları için gerekli çözelti en aza indirmek için, bir hidrofobik bariyer kalemle daire hücreler, daha sonra hemen oda sıcaklığında PBS, 15 dakika içinde 150 ul% 3 paraformaldehit (RT) ile tespit edin. <ol><li> PBS içinde 150 ul% 3 BSA ile 5 dakika boyunca iki kez yıkanır. </li><li> Oda sıcaklığında PBS, 20 dakika içinde% 0.5 Triton-X-100 ile Permeabilize hücreleri. </li><li> 5.1.1 gibi tekrarlayın yıkama adımı. </li></ol></li><li> Edu reaksiyon cockt hazırlayınail kit talimatlarına fazla kullanımdan önce en az 30 daha uygun. <ol><li> Her slayt için 150 ul kokteyl ekle oda sıcaklığında 30 dakika inkübe. </li><li> 5.1.1 gibi tekrarlayın yıkama adımı. </li></ol></li><li> Oda sıcaklığında, 30 dakika PBS içinde 2,000: Nükleer karşıt Hoechst33342 1 seyreltilmiş 150 ul inkübe edin. <ol><li> PBS 1X 150 ul, 5 dakika boyunca iki kez yıkanır. </li></ol></li><li> daha sonra yavaş yavaş, 1-2 hücrelerine montaj orta anti-solmaya damla kabarcıklar önlemek için alt lamel ekleyin. <ol><li> Oda sıcaklığında, karanlıkta gece boyunca tedavi için orta montaj izin verin. </li><li> slayt lamel mühür net oje kullanın. </li></ol></li><li> DAPI (Hoeschst33342) saptayabilen uyarma / emisyon filtreleri ile donatılmış bir floresan mikroskop kullanılarak 40X büyütmede görüntü slaytlar, ve Texas Kırmızı (Alexa Fluor 594). </li></ol>

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The authors declare that they have no competing financial interests

tam kesisi ve fleksör digitorum longus tendon onarım fare modeli için cerrahi prosedür bu çalışmada sunulmuştur. Buna ek olarak sitolojisi santrifüj küçük hücre popülasyonlarının konsantre yeni bir uygulama fleksor tendonu iyileşmesi sırasında hücresel ortamda kantitatif immünositokimyasal analizi için izin gösterilmektedir. Fleksor tendon onarımı Bu model nakavt modeller kullanılarak iyileşme sürecinde değişiklik ya da farmakolojik müdahale ile değerlendirmek için kullanılabilen bir ye...

fleksör önkol kaslarının ve dijital kılıflar ile uyum içinde el işçiliği fleksör tendon basamak fleksiyon ve el kavrama işlevini etkinleştirmek için. Fleksör tendonlar elin palmar uzanmak; Bu nispeten yüzeysel konumu genellikle elle travma sırasında fleksör tendon yaralanmaları ile sonuçlanır. Tendon, normal tendon dokusunun 1 bir skar dokusu tepki yerine rejenerasyon yoluyla iyileşir. Bu skar dokusu tendon sürekliliğini sağlarken, işlev sağlıklı tendon göre azalmış dramatik bir. Tendon-skar dokusu kompozitler kopma olasılığı daha yüksektir tamir tendonları render, bozulmuş mekanik özellikleri 1 ile karakterizedir. Buna ek olarak, yara dokusu, tendon boyutu ve toplu olarak bir artışa neden olur, yerli tendonun kollajen lif yapısının organizasyonu yoksundur. Tendon-kılıf biriminin anatomik kısıtlamalar, tendon boyutunda bile mütevazı bir artış göz önüne alındığında can ölçüde kırmızıkayma tendon fonksiyonunu ve hareket ve el fonksiyonu dolayısıyla haneli aralığını UCE. Fleksor tendonların 1960 yaralanmaları, el II.Bölgede özellikle öncesinde, rutin nedeniyle bu onarım 2 ile ortaya çıkan iyileşme ciddi komplikasyonlara tamir edilmedi. Elin Bu alan &#39;adam hiçbir arazi 3&#39; olarak sevk edildi. Ancak, cerrahi teknikler, dikiş desenleri ve fizik tedavi rehabilitasyon protokolleri gelişmeler dramatik fleksör tendon onarımı 2 sonuçlarını iyileştirilmiştir. Bu ilerlemelere rağmen, onarım% 40&#39;a varan el fonksiyonlarını 4 engellemek için yeterli yapışma oluşumuna neden. Bu nedenle, biyolojik bir iyileşme geliştirmek için gereklidir. Ne yazık ki, çok az hücresel ve moleküler düzeyde tendon iyileşme süreci hakkında bilinmektedir. Bu nedenle, amaç, temel understandin geliştirmek için kullanılabilecek bir sıçan modelinde geliştirmektişifa artırmak için yeni terapötik hedefler belirlemek için bir araç olarak fleksor tendon iyileşmesi ve skar oluşumu yanıt hücresel ve moleküler bileşenleri, g. Büyük hayvan modelleri fleksör tendon iyileşme sürecinin anlaşılmasını ilerletmek vesile olmuştur. Köpek ve tavşan çalışmaları fleksör tendonların 5,6 içsel ve dışsal şifa yeteneği hem de göstermiştir, immobilizasyon 7 adezyon göreceli minimize erken kontrollü pasif hareket önemi yanı sıra, iyileşme sürecine 8 farklı sütür desen etkileri 9. Buna ek olarak, köpek modeli şifa 10 artırmak için öteleme doku mühendisliği yaklaşımları test yararlı olmuştur. Bununla birlikte, görece maliyet sıçangil özel reaktiflerin mevcudiyeti dahil büyük hayvan modeli için bir fare modeli kullanılmaktadır kullanılarak birçok önemli avantajı vardır ve küresel Knoc üretilmesi kolaylığıya da dokuya özel silme / aşırı yapıları aşımlarının k. Ayrıca, fleksor tendonları 11 ile ilgili olarak insan ve fareler arasındaki fonksiyonel benzerlik bir kemirgen modeli geliştirme potansiyel yararını göstermektedir. fleksor tendon transeksiyonu ve onarım taklit bol skar dokusu ve bozulmuş mekanik özellikleri oluşumu da dahil olmak üzere klinik iyileşme birçok yönleri, bir fare modeli geliştirilmesi. Burada açıklanan model onarım sitesini korumak amacıyla nedeniyle myotendinous kavşakta FDL kesilerinden için klinik pratikte gerçek bir tekrarlama değil. Onarım meydana tendon orta kısmını kaplayan herhangi bir sinovyal kılıf yoktur Dahası, bu model, şifa yanıt sinovyal kılıf hücrelerinin katkısı hesaba katmaz. Bu kısıtlamalara rağmen, bu model fare modellerinde daha clos gösterilebilir henüz hareket sınırlayıcı yapışıklıklar üreten aralığı avantajı vardırely klinik senaryo yaklaşmaktadır. Bu model knock-out fare modelleri 12,13 değerlendirmek ve şifa 14-17 geliştirmek için farklı farmakolojik yaklaşımlar test etmek için kullanılır olmuştur. Histolojik immünohistokimyasal kullanarak, bu modelin analizleri ve in situ melezleme, iyileşme sırasında anahtar genlerin ve proteinlerin lokalizasyon için önemli bilgiler sağlayabilir. Ancak, histoloji yalnızca bir kesit mekansal analiz sağlar ve tüm doku boyunca kantifikasyonunu izin vermez. Akış sitometri daha nicel bir yaklaşımı temsil eder, ama hücrelerin sadece çok sınırlı sayıda fare modelinde iyileşen tendon dokusundan izole edilebilir, ve bu sayı daha da tespit, geçirgenleştirme ve yıkama adımları sırasında azaltılır. hesapta bu alarak, sitometri nedeniyle gerekli olacaktır hayvan sayısı bir olanaksız bir yaklaşım olur akar. Alternatif bir yöntem için, bu küçük bir hücre popülasyonunun çoğunluğunu korumak için gerekli olanayrıca şifa ortam karakterize etmek. Burada gösterilen Bunu gerçekleştirmek için kullanılan yöntem, bağışıklık kimyası ve ardından bir cam lam üzerine sitoloji santrifüj yoluyla izole hücrelerin konsantrasyonu içerir. Bu çalışmada edu (5-etinil-2&#39;deoxyuridine bir timidin analoğu) dahil edilmesi ve daha sonra etiket iyileşmesi yerinde hücrelerin göreceli proliferatif durumu belirlemek için kullanılmıştır. Bu yaklaşım, hücre çoğalması, gen nakavt veya aşırı ekspresyonuna farmakolojik tedavilerin etkisini test etmek için, ya da farklı hücre popülasyonu belirlemek ve ölçmek için uygulanabilir.

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murine flexor tendon injury and repair surgery

Tendon neredeyse tüm vücudun hareketini kolaylaştıran, iskelet kas ve kemik bağlar. elinde, fleksör tendonlar (FTS) parmakları ve genel el fonksiyonunun fleksiyonuna etkinleştirin. FT yaralanmaları yaygındır ve tatmin edici iyileşme çoğu zaman aşırı skar dokusu ve tendon ve çevre doku arasındaki yapışıklıklar bozulur. Ancak, küçük FT onarım moleküler ve hücresel bileşenleri hakkında bilinmektedir. Bu amaçla, hareket bozukluğu aralığı dahil olmak üzere, insanlardaki iyileşme birçok açıdan, tekrarlar ve mekanik özellikleri azalma FT tamir bir murin modeli, geliştirilmiş ve daha önce tarif edilmiştir. İşte bu cerrahi işlem derinlemesine bir gösteri kesiye ve fare arka pençesine fleksör digitorum longus (FDL) tendonun sonraki tamir edilmesi suretiyle sağlanır. Bu teknik gen kazanç veya kayıp fonksiyon-etkilerini değerlendirmek, ve verimli test etmek için, farklı hücre tiplerinin soyu analizi yapmak için kullanılabiliriyileşme sürecinde kullanılan ilaç tedavilerinin etkinlik gösterir. i) transeksiyonu ve onarım meydana fare arka ayağın, orta kısmında FDL tendon, bir sinovyal kılıf ile çevrili değildir: Ancak, bu modelin iki temel sınırlamalar vardır. Bu nedenle, bu model skar oluşumu sürecine kılıfın potansiyel katkısı hesaba katmaz. ii) onarım sitenin bütünlüğünü korumak için, FT olasılıkla artmış yara oluşumuna katkıda tendonun mekanik kuvvetleri azalan myotendinous kavşakta serbest bırakılır. akım sitometri analizi için iyileşme sürecinde FT granülasyon dokusu yeterli hücrelerin izolasyonu zor olduğu kanıtlanmıştır; Bu hücrelerin konsantre sitoloji santrifüj kullanılan başka bir yöntemdir, ve imünoflöresan markalama yapılabilir üzerinde hücre preparatlarının oluşturulmasına izin verir. Bu yöntem ile, FT iyileşme sırasında hücreler ya da ilgi duyulan proteinlerin ölçümü mümkün olur.

Fleksör digitorum longus buzağı bulunan (FDL) kas, myotendinous proksimal çalıştığı, (Şekil 1A mavi özetlenen ve Şekil 2A histolojik gösterilen) fleksör tendon aracılığıyla fare arka pençe rakamlarını flex hareket kavşak ve distal falanks sonlandırır. Arka pençe basamağı (kırmızı oklar, Şekil 1A) için çatallanma, proksimal fleksör tendon iyileşme Bu modelde, FDL tendonu nakledilmiştir ve orta dibinde o...

Watch this Scientific Journal Video about Murine Flexor Tendon Injury and Repair Surgery at JoVE.com

Murine Flexor Tendon Injury and Repair Surgery

el fleksör tendonlar sıklıkla bozulmuş el fonksiyonu yol yaralandı. Bununla birlikte, yara izi dokusunu iyileşme yanıtı iyi karakterize değildir. fleksör tendon iyileşmesi bir fare modeli burada gösterilmiştir. Bu model iyileşme sürecinin genel anlayış geliştirmek ve şifa iyileştirmek için tedavi yaklaşımları değerlendirmek.

Murin Flexor Tendon yaralanmaları ve onarım Cerrahisi

Tendon connects skeletal muscle and bone, facilitating movement of nearly the entire body. In the hand, flexor tendons (FTs) enable flexion of the fingers ...

murine-flexor-tendon-injury-and-repair-surgery

Research

JoVE Journal

Medicine

13.0K Görüntüleme.  University of Rochester Medical Center.  el fleksör tendonlar sıklıkla bozulmuş el fonksiyonu yol yaralandı. Bununla birlikte, yara izi dokusunu iyileşme yanıtı iyi karakterize değildir. fleksör tendon iyileşmesi bir fare modeli burada gösterilmiştir. Bu model iyileşme sürecinin genel anlayış geliştirmek ve şifa iyileştirmek için tedavi yaklaşımları değerlendirmek. 

Video: Murine Flexor Tendon Injury and Repair Surgery

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Sıçanlarda Spinal Kord Yaralanma sonrası Akut ve Kronik Dokunsal Duyu Testi

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

Tıp

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

Sonsuz Horizon Darbeli kullanarak tek taraflı Servikal Spinal Kord Yaralanması bir Contusive Modeli

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

Endovasküler Aort Anevrizma Onarımı Yeni Bir murin Modeli

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Travmatik Beyin Hasarı için Kontrollü Kortikal Etki Modeli

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Mouse Fetal Skin Model of Scarless Wound Repair

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs across species but is age dependent. The transition from a scarless to scarring phenotype occurs in the third trimester of pregnancy in humans and around embryonic day 18 (E18) in mice. However, this varies with the size of the wound with larger defects generating a scar at an earlier gestational age. The emergence of lineage tracing and other genetic tools in the mouse has opened promising new avenues for investigation of fetal scarless wound healing. However, given the inherently high rates of morbidity and premature uterine contraction associated with fetal surgery, investigations of fetal scarless wound healing in vivo require a precise and reproducible surgical model. Here we detail a reliable model of fetal scarless wound healing in the dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

During mammalian development, early gestational skin wounds heal without a scar. Here we detail a reliable and reproducible model of fetal scarless wound healing in the cutaneous dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

Scarless Yara Repair Fare Fetal Cilt Modeli

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

Arter Restenozun bir fare modeli: Femur Tel Yaralanma Teknik Yönleri

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Bağırsak İskemi-reperfüzyon Hasarı Murin Modeli

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Bir Fare Modelinde İnfrarenal Abdominal Aort Bağlama Ekipmanları tarafından tekrarlanabilir Arter denüdasyon Yaralanma

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

Evaluation of Stem Cell Therapies in a Bilateral Patellar Tendon Injury Model in Rats

Regenerative medicine provides novel alternatives to conditions that challenge traditional treatments. The prevalence and morbidity of tendinopathy across species, combined with the limited healing properties of this tissue, have prompted the search for cellular therapies and propelled the development of experimental models to study their efficacy. Umbilical cord matrix-derived mesenchymal stem cells (UCM-MSC) are appealing candidates because they are abundant, easy to collect, circumvent the ethical concerns and risk of teratoma formation, yet resemble primitive embryonic stem cells more closely than adult tissue-derived MSCs. Significant interest has focused on chitosan as a strategy to enhance the properties of MSCs through spheroid formation. This paper details techniques to isolate UCM-MSCs, prepare spheroids on chitosan film, and analyze the effect of spheroid formation on surface marker expression. Consequently, creation of a bilateral patellar tendon injury model in rats is described for in vivo implantation of UCM-MSC spheroids formed on chitosan film. No complication was observed in the study with respect to morbidity, stress rising effects, or tissue infection. The total functional score of the operated rats at 7 days was lower than that of normal rats, but returned to normal within 28 days after surgery. Histological scores of tissue-healing confirmed the presence of a clot in treated defects evaluated at 7 days, absence of foreign body reaction, and progressing healing at 28 days. This bilateral patella tendon defect model controls inter-individual variation via creation of an internal control in each rat, was associated with acceptable morbidity, and allowed detection of differences between untreated tendons and treatments.

This paper describes the preparation and evaluation of umbilical cord matrix-derived mesenchymal stem cells spheroids with a bilateral patellar tendon defect model in a rat. This model was associated with an acceptable morbidity and was found to detect differences between untreated and treated tendons, and between the two treatments tested.

Kök hücre tedavileri Sıçanlarda bir ikili Patellar Tendon yaralanma modelinde değerlendirilmesi

Regenerative medicine provides novel alternatives to conditions that challenge traditional treatments. The prevalence and morbidity of tendinopathy across ...

Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats

Incomplete spinal cord injury (SCI) often leads to impairments of sensorimotor functions and is clinically the most frequent type of SCI. Human Brown-S&#233;quard syndrome&#160;is a common type of incomplete SCI caused by a lesion to one half of the spinal cord which results in paralysis and loss of&#160;proprioception&#160;on the same (or ipsilesional) side as the injury, and loss of pain and temperature sensation on the opposite (or contralesional) side. Adequate methodologies for producing a spinal cord lateral hemisection (HX) and assessing neurological impairments are essential to establish a reliable animal model of Brown-S&#233;quard syndrome. Although lateral hemisection model plays a pivotal role in basic and translational research, standardized protocols for creating such a hemisection and assessing unilateralized function are lacking. The goal of this study is to describe step-by-step procedures to produce a rat spinal lateral HX at the 9th thoracic (T9) vertebral level. We, then, describe a combined behavior scale for HX (CBS-HX) that provides a simple and sensitive assessment of asymmetric neurological performance for unilateral SCI. The CBS-HX, ranging from 0 to 18, is composed of 4 individual assessments which include unilateral hindlimb stepping (UHS), coupling, contact placing, and grid walking. For CBS-HX, the ipsilateral and contralateral hindlimbs are assessed separately. We found that, after a T9 HX, the ipsilateral hindlimb showed impaired behavior function whereas the contralateral hindlimb showed substantial recovery. The CBS-HX effectively discriminated behavioral functions between ipsilateral and contralateral hindlimbs and detected temporal progression of recovery of the ipsilateral hindlimb. The CBS-HX components can be analyzed separately or in combination with other measures when needed. Although we only provided visual descriptions of the surgical procedures and behavioral assessments of a thoracic HX, the principle may be applied to other incomplete SCIs and at other levels of the injury.

Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury.