Yazarlar Lena Brix, V.M.D, Hayvan Araştırmaları Enstitüsü, Hannover Tıp Fakültesi'nin yanı sıra Bayan Jutta Dammann'a, Araştırma Hayvan Bakım Tesisi, Hamburg-Eppendorf Üniversitesi Tıp Merkezi, Almanya'ya, ameliyat öncesi ve perioperatif hayvan bakımı ve hayvan işleme konusundaki teknik yardımları için teşekkür ediyor. Yazarlar ayrıca, Lüksemburg'daki Hôpital Kirchberg Vasküler Cerrahi Bölümü'nden Dr. Daniel Manzoni'ye teknik yardımı için teşekkür eder.

Constantin J.C. Trepte, Maquet'in dersleri için onur ödülü aldı. Diğer tüm yazarlar çıkar çatışması olmadığını beyan eder. Bu çalışma Avrupa Anesteziyoloji Derneği Genç Araştırmacı Start-Up Grant 2018 tarafından desteklendi.

Omurilik iskemisi tarafından indüklenen SCI, hasta sonucu 1 , 2 , 3 , 4,10,11,12üzerinde muazzam bir etkisi olan aort onarımının önemli bir komplikasyonudur. SCI'yı önlemek ve tedavi etmek için mikrosilkülasyon hedefli tedaviler en umut verici olanıdır. Protokol, gerçek zamanl?...

İskemi/reperfüzyon (SCI) ile indüklenen omurilik hasarı, aort onarımınınazaltılmış sonuç 1 , 2,3,4ile ilişkili en yıkıcı komplikasyonlarından biridir. SCI için mevcut önleme ve tedavi seçenekleri arasında makrohemodynamik parametrelerin optimizasyonu ve omurilik perfüzyon basıncını iyileştirmek için beyin omurilik sıvı basıncının (CSP) normalleştirilmesi 2 ,5,6,7,8,9bulunur. Bu manevraların uygulanmasına rağmen, SCI insidansı hala aort onarımının karmaşıklığına bağlı olarak% 2 ila%31arasında değişmektedir 10,11,12.
Son zamanlarda, mikrosirkülasyon artan dikkat kazanmıştır13,14. Mikroskrkülasyon hücresel oksijen alımı ve metabolik değişim alanıdır ve bu nedenle organ fonksiyonu ve hücresel bütünlükte kritik bir rol oynar13. Bozulmuş mikrosirkülatör kan akışı, artan mortalite15 , 16 , 17 , 18,19ile ilişkili doku iskemisinin önemli bir belirleyicisidir. Omurilik mikrosirkülasyonunun bozulması, nörolojik fonksiyonun azalması ve sonucu20 , 21,22,23ile ilişkilidir. Bu nedenle, SCI tedavisi için mikroperfüzyonun optimizasyonu en umut verici bir yaklaşımdır. Makrosikülatör optimizasyonuna rağmen mikro sirkülatör bozuklukların kalıcılığı26 , 27,28,29olarak tanımlanmıştır. Bu hemodinamik tutarlılık kaybı, iskemi/reperfüzyon dahil olmak üzere çeşitli koşullarda sıklıkla meydana gelir ve doğrudan mikro sirkülasyon hedefli tedavilere duyulan ihtiyacı vurgular26,27,30.
Şimdiye kadar, sadece birkaç çalışma omurilik mikro sirkülatör davranışının gerçek zamanlı değerlendirilmesi için lazer-Doppler probları kullandı20,31. Mevcut çalışmalar genellikle aralıklı kullanım ve ölüm sonrası analiz ile sınırlı olan mikroküreci enjeksiyon tekniklerini kullanmıştır32,33. Mikroküreç enjeksiyon tekniği kullanılarak yapılan farklı ölçümlerin sayısı, farklı dalga boylarına sahip mikrokürelerin mevcudiyeti ile sınırlıdır. Ayrıca, Lazer-Doppler tekniklerinin aksine, bu yöntem için ölüm sonrası doku işleme ve analize ihtiyaç duyulduğu için mikroperfüzyonun gerçek zamanlı olarak değerlendirilmesi mümkün değildir. Burada, bir porcine büyük hayvan modeli olan iskemi/reperfüzyonda omurilik mikroskrkülasyonunun gerçek zamanlı olarak değerlendirilmesi için deneysel bir protokol sunuyoruz.
Bu çalışma, kristaloidlerin ve kolloidlerin iskemi/reperfüzyondaki mikroserkülasyon üzerindeki etkisini karşılaştıran randomize bir çalışmanın yanı sıra sıvıların ve vazopresörlerin omurilik mikroperfüzyonu üzerindeki etkileri üzerine yapılan keşifsel randomize bir çalışmanın bir parçasıydı. Akış probu 2 noktalı kalibrasyon ve basınç ucu kateter kalibrasyonu daha öncetanımlanmıştır 34. Bildirilen protokole ek olarak, floresan mikroküreler, daha önce açıklandığı gibi, her hayvan için 12 omurilik dokusu örneği kullanılarak, üst omuriliği temsil eden 1-6 ve alt omuriliği temsil eden 7-12 örnekleri kullanılarak omurilik mikroperfüzyonunun ölçümü için kullanılmıştır35,36. Lazer-Doppler kayıtları ve makrohemodynamik değerlendirmenin tamamlanmasından sonra her ölçüm adımı için mikrosfer enjeksiyonu yapıldı. Histopatolojik değerlendirme daha önce açıklandığı gibi Kleinman-Score kullanılarak yapıldı37.

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			<td>CardioMed Flowmeter</td>
			<td>Medistim AS, Oslo, Norway</td>
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			<td>Smiths Medical Germany GmbH, Grasbrunn, Germany</td>
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			<td>Infinity Delta</td>
			<td>Dr&#228;ger Medical, L&#252;beck, Germany</td>
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			<td>Dr&#228;ger Medical, L&#252;beck, Germany</td>
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			<td>ADInstruments Ltd., Oxford, UK</td>
			<td>v8.1.16</td>
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			<td>LiquoGuard 7</td>
			<td>M&#246;ller Medical GmbH, Fulda, Germany</td>
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			<td>Millar Micro-Tip Pressure Catheter (5F, Single, Curved, 120cm, PU/WD)</td>
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			<td>moor VMS LDF</td>
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			<td>moor VMS Research Software</td>
			<td>moor Instruments, Devon, UK</td>
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			<td>Perivascular Flow Module</td>
			<td>Transonic Systems Inc., Ithaca, NY, USA</td>
			<td>TS 420</td>
			<td>Flow-Module for Flow-Probe Aorta</td>
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			<td>PiCCO 2, Science Version</td>
			<td>Getinge AB, G&#246;teborg, Sweden</td>
			<td>v. 6.0</td>
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			<td>PiCCO 5 Fr. 20cm</td>
			<td>Getinge AB, G&#246;teborg, Sweden</td>
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			<td>Thermistor-tipped Arterial Line&#160;</td>
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			<td>PowerLab</td>
			<td>ADInstruments Ltd., Oxford, UK</td>
			<td>PL 3516</td>
			<td>Synchronic Laser-Doppler, Blood Pressure, ECG and Blood-Flow Aquisition Hardware</td>
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			<td>QuadBridgeAmp</td>
			<td>ADInstruments Ltd., Oxford, UK</td>
			<td>FE 224</td>
			<td>Four Channel Bridge Amplifier for Laser-Doppler and Invasive Blood Pressure Aquisition</td>
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			<td>Silverline</td>
			<td>Spiegelberg, Hamburg, Germany</td>
			<td>ELD33.010.02</td>
			<td>Cerebrospinal Fluid Drainage</td>
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			<td rowspan="2">SPSS statistical software package&#160;</td>
			<td rowspan="2">IBM SPSS Statistics Inc., Armonk, New York, USA</td>
			<td rowspan="2">v. 27</td>
			<td rowspan="2">Statistical Software</td>
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			<td>Twinwarm Warming System</td>
			<td>Moeck &#38; Moeck GmbH, Hamburg, Germany</td>
			<td>12TW921DE</td>
			<td>Warming System</td>
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			<td>Universal II Warming Blanket</td>
			<td>Moeck &#38; Moeck GmbH, Hamburg, Germany</td>
			<td>906</td>
			<td>Warming Blanket</td>
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			<td>VP 3 Probe, 8mm length (individually manufactured)</td>
			<td>moor Instruments, Devon, UK</td>
			<td></td>
			<td>Laser-Doppler Probe</td>
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			<td>Zeus</td>
			<td>Dr&#228;ger Medical, L&#252;beck, Germany</td>
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			<td>Anesthesia Machine</td>
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real-time assessment of spinal cord microperfusion in a porcine model of ischemia/reperfusion

Omurilik yaralanması aort onarımının yıkıcı bir komplikasyonudur. Omurilik hasarının önlenmesi ve tedavisine yönelik gelişmelere rağmen, görülme sıklığı hala oldukça yüksektir ve bu nedenle hasta sonucunu etkiler. Mikroskrkülasyon doku perfüzyonu ve oksijen temininde önemli bir rol oynar ve genellikle makrohemodynamiklerden ayrışır. Bu nedenle, omurilik mikroskrkülasyonunun doğrudan değerlendirilmesi, mikrosilkülasyon hedefli tedavilerin geliştirilmesi ve omurilik mikroskrkülasyonu ile ilgili mevcut yaklaşımların değerlendirilmesi için gereklidir. Bununla birlikte, yöntemlerin çoğu omurilik mikroskrkülasyonunun gerçek zamanlı değerlendirmesini sağlamaz. Bu çalışmanın amacı, doğrudan omuriliğe yerleştirilen lazer-Doppler iğne probları kullanılarak gerçek zamanlı omurilik mikrosikülatör değerlendirmesi için standartlaştırılmış bir protokolü tanımlamaktır. Omurilik mikroserkülasyonunun bozulmasını sağlamak için bir porcine iskemi/reperfüzyon modeli kullandık. Ayrıca floresan mikrokürez enjeksiyon tekniği kullanılmıştır. Başlangıçta, hayvanlar uyuşturuldu ve mekanik olarak havalandırıldı. Daha sonra lazer-Doppler iğne probu yerleştirilmesi, ardından beyin omurilik sıvısı drenajının yerleştirilmesi gerçekleştirildi. Aort çapraz kenetleme yapmak için alçalan aortun maruz kalması için ortanca sternotomi yapıldı. İskemi/reperfüzyon, toplam 48 dk supra-çölyak aort çapraz kenetlenmesi ile indüklendi, ardından reperfüzyon ve hemodinamik stabilizasyon yapıldı. Lazer-Doppler Flux makrohemodynamik değerlendirmeye paralel olarak gerçekleştirildi. Ek olarak, stabil bir beyin omurilik basıncını korumak için otomatik beyin omurilik sıvı drenajı kullanılmıştır. Protokolün tamamlanmasının ardından hayvanlar kurban edildi, histopatolojik ve mikrosfer analizi için omurilik hasadı yapıldı. Protokol, lazer-Doppler probları kullanılarak omurilik mikroperfüzyon ölçümlerinin fizibilitesini ortaya koymakta ve iskemi sırasında belirgin bir azalmanın yanı sıra reperfüzyon sonrası iyileşme göstermektedir. Sonuçlar floresan mikroküreci değerlendirme ile karşılaştırılabilir davranış gösterdi. Sonuç olarak, bu yeni protokol iskemi/ reperfüzyon koşullarında gerçek zamanlı omurilik mikroperfüzyon değerlendirmesi kullanılarak gelecekteki çalışmalar için yararlı bir büyük hayvan modeli sağlayabilir.

Altı hayvan da protokolün tamamlanmasına kadar hayatta kaldı. Hayvan ağırlığı 48.2 ± 2.9 kg; beş hayvan erkekti ve bir hayvan dişiydi. Omurilik iğnesi probu takılması ve omurilik Flux ölçümü tüm hayvanlarda mümkündü.
İskemi indüksiyonu için aort çapraz kenetleme sırasında ve unclamping ve reperfüzyon sırasında serebral mikro sirkülatör ve makrohemodynamik kayıtlarla birlikte gerçek zamanlı om...

Watch this Scientific Journal Video about Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion at JoVE.com

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Omurilik mikroskrkülasyonu omurilik yaralanmasında önemli bir rol oynar. Çoğu yöntem, mikroskrkülasyon hedefli tedavilerin geliştirilmesi için gerekli olan omurilik mikroskrkülasyonunun gerçek zamanlı değerlendirilmesine izin vermez. Burada, büyük bir iskemi /reperfüzyon hayvan modelinde Lazer-Doppler-Akış İğne probları kullanarak bir protokol öneriyoruz.

İskemi/Reperfüzyon Porsin Modelinde Omurilik Mikroperfüzyonunun Gerçek Zamanlı Değerlendirilmesi

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

real-time-assessment-spinal-cord-microperfusion-porcine-model

Research

JoVE Journal

Medicine

3.4K Görüntüleme.  University Medical Center Hamburg-Eppendorf. Omurilik mikroskrkülasyonu omurilik yaralanmasında önemli bir rol oynar. Çoğu yöntem, mikroskrkülasyon hedefli tedavilerin geliştirilmesi için gerekli olan omurilik mikroskrkülasyonunun gerçek zamanlı değerlendirilmesine izin vermez. Burada, büyük bir iskemi /reperfüzyon hayvan modelinde Lazer-Doppler-Akış İğne probları kullanarak bir protokol öneriyoruz.

Video: Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

Normotermik Kardiyak Arrest ve Kardiyopulmoner Resusitasyon: İskemi-Reperfüzyon Hasarı Fare Modeli

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

Tıp

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 ...

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, intestinal ischemia as well as following transplant and cardiovascular surgery.1 Reperfusion of previously ischemic tissue, while essential for the prevention of irreversible tissue injury, elicits excessive inflammation of the affected tissue. Adjacent to the production of reactive oxygen species, activation of the complement system and increased microvascular permeability, the activation of leukocytes is one of the principle actors in the pathological cascade of inflammatory tissue damage during reperfusion.2, 3 Leukocyte activation is a multistep process consisting of rolling, firm adhesion and transmigration and is mediated by a complex interaction between adhesion molecules in response to chemoattractants such as complement factors, chemokines, or platelet-activating factor.4
While leukocyte rolling in postcapillary venules is predominantly mediated by the interaction of selectins5 with their counter ligands, firm adhesion of leukocytes to the endothelium is selectin-controlled via binding to intercellular adhesion molecules (ICAM) and vascular cellular adhesion molecules (VCAM).6, 7 
Gold standard for the in vivo observation of leukocyte-endothelial interaction is the technique of intravital microscopy, first described in 1968.8
Though various models of IRI (ischemia-reperfusion injury) have been described for various organs, 9-12 only few are suitable for direct visualization of leukocyte recruitment in the microvascular bed on a high level of image quality.8 
We here promote the digital intravital epifluorescence microscopy of the postcapillary venule in the cremasteric microcirculation of the rat 13 as a convenient method to qualitatively and quantitatively analyze leukocyte recruitment for IRI-research in striated muscle tissue and provide a detailed manual for accomplishing the technique. We further illustrate common pitfalls and provide useful tips which should enable the reader to truly appreciate, and safely perform the method.
In a step by step protocol we depict how to get started with respiration controlled anesthesia under sufficient monitoring to keep the animal firmly anesthetized for longer periods of time. We then describe the cremasteric preparation as a thin flat sheet for outstanding optical resolution and provide a protocol for leukocyte imaging in IRI that has been well established in our laboratories.

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Digital intravital epifluorescence microscopy of postcapillary venules in the cremasteric microcirculation is a convenient method to gain insights into leukocyte-endothelial interaction in vivo in ischemia-reperfusion injury (IRI) of striated muscle tissue. We here provide a detailed protocol to safely perform the technique and discuss its applications and limitations.

Sıçan Cremaster&#39;ın kas iskemi-reperfüzyon Hasarı içinde Lökosit-endotelyal Etkileşim Gerçek zamanlı Dijital Görüntüleme (IRI)

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

Sıçanlarda Şiddetli Spinal Kord Yaralanması bir bere modeli

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Background 
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered advancements to decrease the occurrence this injury.&#160;The current model demonstrates reproducible lower&#160;extremity paralysis following thoracic aortic occlusion.
Methods 
Adult male C57BL6 mice were anesthetized with isoflurane. Through a cervicosternal incision the aorta was exposed. The descending thoracic aorta and left subclavian arteries were identified without entrance into pleural space. Skeletonization of these arteries was followed by immediate closure (Sham) or occlusion for 4 min (moderate ischemia) or 8 min (prolonged ischemia). The sternotomy and skin were closed and the mouse was transferred to warming bed for recovery.&#160; Following recovery, functional analysis was obtained at 12 hr intervals until 48 hr.
Results 
Mice that underwent sham surgery showed no observable hind&#160;limb deficit. Mice subjected to moderate ischemia for 4 min had minimal functional deficit at 12 hr followed by progression to complete paralysis at 48 hr. Mice subjected to prolonged ischemia had an immediate paralysis with no observable hind-limb movement at any point in the postoperative period. There was no observed intraoperative or post&#160;operative mortality.
Conclusion 
Reproducible lower extremity paralysis whether immediate or delayed can be achieved in a murine model. Additionally, by using a median sternotomy and careful dissection, high survival rates, and reproducibility can be achieved.

The lack of mechanistic understanding of spinal cord ischemia-reperfusion injury has hindered further adjuncts to prevent paraplegia following high risk aortic operations. Thus, the development of animal models is imperative. This manuscript demonstrates reproducible lower extremity paralysis following thoracic aortic occlusion in a murine model.

Spinal Kord İskemi-reperfüzyon bir Mürin Modelinde Torasik Aort Tıkanıklığı tarafından tekrarlanabilir Parapleji

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

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 Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

A Radyo-telemetrik Sistemi Omurilik transeksiyonu ve Embriyonik Nöral Hücre Greftler Kök Sıçanlarda Kardiyovasküler Fonksiyonu Monitör

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

Deneysel Spinal Kord Yaralanmalarında Omurilik Kan Akımının Değerlendirilmesi için geliştirilmiş Ultrason Görüntüleme Kontrast

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

Langendorff-perfüze sıçan Hearts indüksiyon ve İskemi-reperfüzyon Hasarı Değerlendirilmesi

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

Bir Yenidoğan Fare Omurilik Sıkıştırma Yaralanması Modeli

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...