Name: angiogenesis in the ischemic rat lung
Uploaded: 2013-02-08T13:00:00
Description: Watch this Scientific Journal Video about Angiogenesis in the Ischemic Rat Lung at JoVE.com

Reconocemos la labor del Dr. Adlah Sukkar, MD para ayudar con la colada del pulmón. Este trabajo ha sido financiado por el NHLBI, HL088005.

Todos los protocolos realizados en ratas han sido aprobados por el Cuidado de Animales Universidad Johns Hopkins y el empleo, y de conformidad con las directrices del NIH. Siempre que sea posible, el animal debe ser preparado quirúrgicamente en un área separada de la zona quirúrgica para minimizar la contaminación de la zona quirúrgica. 1. La anestesia / analgesia <ol><li> Rata lugar (ratas Sprague Dawley macho, 125-150 g; Harlan, Indianapolis, IN) en una cámara de inducción infundi...

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Med. 18, 507-522 (1913).</li><li>Endrys, J., Hayat, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+bronchopulmonary+collaterals+and+collateral+blood+flow+in+patients+with+chronic+thromboembolic+and+primary+pulmonary+hypertension.">Comparison of bronchopulmonary collaterals and collateral blood flow in patients with chronic thromboembolic and primary pulmonary hypertension.</a> Heart. 78, 171-176 (1997).</li><li>Virchow, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uber+die+Standpunkte+in+den+Wissenschaftlichen+Medizin.">Uber die Standpunkte in den Wissenschaftlichen Medizin.</a> Virchow Archiv. 1, 1-19 .</li><li>Fadel, E., Mazmanian, G. 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=Lung+reperfusion+injury+after+chronic+or+acute+unilateral+pulmonary+artery+occlusion.">Lung reperfusion injury after chronic or acute unilateral pulmonary artery occlusion.</a> Am. J. Respir. Crit. Care Med. 157, 1294-1230 (1998).</li><li>Charan, N. B., Carvalho, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiogenesis+in+bronchial+circulatory+system+after+unilateral+pulmonary+artery+obstruction.">Angiogenesis in bronchial circulatory system after unilateral pulmonary artery obstruction.</a> J. Appl. 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Physiol. 110, 538-544 (2011).</li><li>Remy-Jardin, M., Bouaziz, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bronchial+and+nonbronchial+systemic+arteries+at+multi-detector+row+CT+angiography:+comparison+with+conventional+angiography.">Bronchial and nonbronchial systemic arteries at multi-detector row CT angiography: comparison with conventional angiography.</a> Radiology. 233, 741-749 (2004).</li><li>Baluk, P., Tammela, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+persistent+lymphatic+vessel+hyperplasia+in+chronic+airway+inflammation.">Pathogenesis of persistent lymphatic vessel hyperplasia in chronic airway inflammation.</a> J. Clin. Invest. 115, 247-257 (2005).</li><li>Bailey, S. 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La ligadura de la arteria pulmonar izquierda en todas las especies estudiadas conduce a la neovascularización robusto sistémica del pulmón isquémica. Hemos presentado los detalles de la técnica quirúrgica en un modelo de rata. Nuestros resultados vascular producido por fundición, histopatología, y en el etiquetado in vivo demuestran que las arterias bronquiales proliferan y perfundir el parénquima pulmonar. Por lo tanto, los mecanismos de la angiogénesis bronquial puede ser estudiada en un mo...

Angiogénesis sistémica en el pulmón está bien reconocida. En estados de enfermedad tales como el asma 2, fibrosis pulmonar intersticial 3, 4 cáncer y tromboembolismo pulmonar crónico 5, la vasculatura sistémica en y alrededor de los prolifera pulmonares e invade el parénquima pulmonar. Sin embargo, los modelos animales para el estudio de esta activación diferencial de la sistémico en lugar de la circulación pulmonar son pocos. Quizás el modelo más reproducible de la neovascularización sistémica en el pulmón del mamífero adulto es el que ocurre después de la inducción de isquemia crónica de la arteria pulmonar. La respuesta a la obstrucción de la arteria pulmonar en seres humanos 5-7, 8 perros, cerdos, ovejas 9 10, 11 conejillos de indias, ratas 1, 12, 13, 14 y ratones es la rápida proliferación de la arteria bronquial, así como las arterias intercostales. Los mecanismos responsables de neovascularización sistémicación del pulmón después de la isquemia pulmonar son en gran parte desconocido y no se han estudiado ampliamente. El curso temporal de la angiogénesis bronquial en la rata después de la obstrucción de la arteria pulmonar se ha descrito cuidadosamente en el trabajo histológico de Weibel 1. La extensión de este trabajo en la rata, nuestro laboratorio ha centrado tanto en los factores de crecimiento importantes en este proceso, así como el resultado fisiológico de este neovasculatura en el pulmón. Los resultados demuestran la quimiocina CXC CINC-3 se eleva temprano después de la isquemia y el tratamiento de ratas con un anticuerpo neutralizante para CXCR2, el receptor de CINC-3, atenúa la angiogénesis 13. La vasculatura recientemente establecida bronquial 14 días después de la aparición de la isquemia pulmonar ha demostrado ser anormal con permeabilidad proteína aumentó significativamente 15. La función pulmonar izquierda no mostraba normal de disminución de la capacidad de difusión y una disminución en el volumen pulmonar 15. Aunque la neovasculatura puede tener contributed a la preservación de tejido pulmonar durante la isquemia pulmonar crónica, parece no ser normal y puede contribuir a una disminución sostenida de la función pulmonar. Quizás uno de los aspectos más curiosos de este modelo se refiere a la distribución espacial de la proliferación de vasos sanguíneos. A pesar de la liberación de factores de crecimiento dentro del parénquima pulmonar por isquemia, la neovasculatura origina en relativamente grandes arterias bronquiales aguas arriba. La arteria bronquial normal surge como una pequeña rama de la aorta e invade el árbol de la vía aérea en la carina. Así, el mecanismo por el cual los factores de crecimiento inducen la fase inicial de la arteriogénesis no está claro. Sugerimos que la rata, con una anatomía vascular similares a los humanos, proporciona una oportunidad única para estudiar los mecanismos responsables de la angiogénesis pulmonar durante la isquemia sistémica. Aunque la obstrucción completa de la arteria pulmonar izquierda es un fenómeno poco frecuente en sujetos humanos,aumento de la vascularidad bronquial parece estar inducida en los pacientes de manera similar cualquiera que sea el sitio y el tamaño de la obstrucción de la arteria pulmonar 16. Por lo tanto, ofrecemos una descripción detallada de la técnica quirúrgica para ligar la arteria pulmonar izquierda en ratas y un medio para cuantificar la magnitud de la angiogénesis.

<table border="1"><tbody><tr><td> Reactivos: </td><td> Empresa </td><td></td><td></td></tr><tr><td> hidrocloruro de buprenorfina, Puralube </td><td> Mayordomo Schein </td><td></td><td></td></tr><tr><td> bupivicaína </td><td> APP Pharmaceuticals </td><td></td><td></td></tr><tr><td> Povidona Yodada bastoncillo con hisopo </td><td> Dynarex Corporación </td><td></td><td></td></tr><tr><td> polipropileno tamaño de la sutura 6-0, 3/8 círculo inverso aguja de corte </td><td> Myco Médico </td><td></td><td></td></tr><tr><td> PE20 tubo </td><td> Becton Dickinson </td><td></td><td></td></tr><tr><td> 15 micras fluoroesferas de poliestireno carmesí </td><td> Invitrogen </td><td></td><td></td></tr><tr><td> 1 ml vidrio de la jeringa Hamilton </td><td> Hamilton Company </td><td> &lt;/ Td&gt; <td></td></tr><tr><td> Equipamiento: </td><td></td><td></td><td></td></tr><tr><td> Genie Plus bomba de jeringa </td><td> Kent Científico </td><td></td><td></td></tr><tr><td> Espectrofotómetro de Fluorescencia </td><td> Digilab </td><td></td><td></td></tr><tr><td> Rodent Ventilator Modelo 683 </td><td> Harvard Apparatus </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> Tabla 1. Tabla de reactivos y equipos específicos. </td></tr></tbody></table>

angiogenesis in the ischemic rat lung

El pulmón adulto se perfunde por tanto la arteria sistémica bronquial y el retorno venoso todo fluye a través de las arterias pulmonares. En la mayoría de las patologías de pulmón, que es más pequeño de la vasculatura sistémica que responde a una necesidad de mejorar la perfusión pulmonar y muestra neovascularización robusto. Isquemia vascular pulmonar inducido por la obstrucción de la arteria pulmonar se ha demostrado que resulta en una rápida angiogénesis arterial sistémica en el hombre, así como en varios modelos animales. Aunque la evaluación histológica de la evolución temporal de la proliferación de la arteria bronquial en ratas se ha descrito cuidadosamente por Weibel 1, los mecanismos responsables de este crecimiento organizado de nuevos vasos no son claras. Ofrecemos detalles quirúrgicos de la inducción de isquemia arteria pulmonar en la rata que lleva a la neovascularización bronquial. La cuantificación de la extensión de la angiogénesis constituye un desafío adicional debido a la presencia de los dos lechos vasculares en el pulmón. Métodospara determinar la angiogénesis funcional basado en inyecciones de microesferas marcadas se proporcionan.

Reparto vascular: Resultados de los efectos de la isquemia arteria pulmonar izquierda en la rata se muestra en la Figura 1. Se muestra un reparto de metacrilato de la vasculatura bronquial y la vascularización extensa del árbol de las vías respiratorias izquierda 28 días después de LPAL. Para obtener este molde, la vasculatura sistémica se inyectó con una mezcla de metacrilato (rojo), retrógrada en la aorta descendente y de la tráquea se canuló y se inyecta con un material a base de silicona b...

Watch this Scientific Journal Video about Angiogenesis in the Ischemic Rat Lung at JoVE.com

Angiogenesis in the Ischemic Rat Lung

El pulmón se perfundieron por tanto la arteria sistémica bronquial y las arterias pulmonares. En la mayoría de las patologías de pulmón, que es más pequeño de la vasculatura sistémica que muestra neovascularización robusto. La cesación del flujo sanguíneo pulmonar promueve la angiogénesis bronquial a paso ligero. Ofrecemos detalles quirúrgicos de la inducción de isquemia arteria pulmonar que promueve la neovascularización bronquial.

La angiogénesis en el pulmón de rata isquémico

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

Research

JoVE Journal

Medicine

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

Multimodal de imágenes de la angiogénesis en un modelo de rata desnuda de las metástasis óseas del cáncer de pecho utilizando imágenes de resonancia magnética, tomografía computarizada y la ecografía volumétrica

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Investigación

Medicina

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

Desprendimiento de Retina modelo en roedores por inyección subretiniana de Hialuronato de Sodio

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

El implante de gel de fibrina en pulmón de ratón para el Estudio de la angiogénesis pulmón específico

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

Método de aislados<em&gt; Ex Vivo</em&gt; Perfusión pulmonar en un modelo de rata: Lecciones Aprendidas de Desarrollo de un Programa EVLP Rata

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

Rata Modelo de neuropatía óptica isquémica posterior inducida por Fotoquímicamente

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

Rata heterotópico abdominal Corazón / Trasplante de un solo pulmón en una configuración de volumen cargado

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

Demostración de la rata isquémica de la piel herida Modelo

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

Un modelo de trasplante de pulmón de rata de lesión por isquemia/reperfusión caliente: optimizaciones para mejorar los resultados

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

Securing the Rat in the Rat Jacket for Electroacupuncture

Wear a pair of gloves before proceeding to fit and secure the rat in the custom made rat jacket. Then using one hand hold the rat jacket's middle and back portion on the table. Using the other hand, gently hold the rat by its tail to pick it up from the feeding cage and place it into the jacket as the rat moves forward into the jacket and its head reaches the headgear point.Using both hands, carefully pull out its left and right upper limbs from the holes in the jacket. Securely wrap the tying ropes from the opening end of the jacket around the rat's trunk, and tie them tightly at the opposite end to adequately restrain its upper limbs. Tie the rope loose enough to prevent hypoxia or local limb ischemia, and tight enough to prevent the rat's escape.In a similar manner, secure its lower limbs.

Asegurar a la rata en la chaqueta de la rata para la electroacupuntura

<meta charset="utf8"></head><body>Monitorización en tiempo real de la dinámica celular en el trasplante de pulmón de ratón

Intravital Two-Photon Microscopy of the Transplanted Mouse Lung

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by crosstalk between donor and recipient cells. A better understanding of these processes relies on the use of preclinical animal models and is aided by an ability to study intra-graft immune cell trafficking in real-time. Intravital two-photon microscopy can be used to image tissues and organs for depths up to several hundred microns with minimal photodamage, which affords a great advantage over single-photon confocal microscopy. Selective use of transgenic mice with promoter-specific fluorescent protein expression and/or adoptive transfer of fluorescent dye-labeled cells during intravital two-photon microscopy allows for the dynamic study of single cells within their physiologic environment. Our group has developed a technique to stabilize mouse lungs, which has enabled us to image cellular dynamics in na&#239;ve lungs and orthotopically transplanted pulmonary grafts. This technique allows for detailed assessment of cellular behavior within the vasculature and in the interstitium, as well as for examination of interactions between various cell populations. This procedure can be readily learned and adapted to study immune mechanisms that regulate inflammatory and tolerogenic responses after lung transplantation. It can also be expanded to the study of other pathogenic pulmonary conditions.

<meta charset="utf8"></head><body>Autor destacado: Avance de la inmunología del trasplante de pulmón a través de imágenes intravitales

Here, we present a protocol to intravitally image the transplanted mouse left lung using two-photon microscopy. This represents a valuable tool for studying cellular dynamics and interactions in real-time following murine lung transplantation.

Microscopía intravital de dos fotones del pulmón de ratón trasplantado

Complications after lung transplantation are largely related to the host immune system responding to the graft. Such immune responses are regulated by ...