The authors would like to acknowledge the Cincinnati Children&#8217;s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.

Declaração uso Animal:: NOTA Todos os procedimentos deste estudo foram realizados com a aprovação e seguir as orientações da Institutional Uso de Animais e do Comitê Care (IACUC) no Hospital Infantil de Cincinnati. Oito semanas de idade C57BL / 6J machos foram obtidos a partir de Jackson Laboratories (Bar Harbor, ME) e deixados a aclimatar durante uma semana antes da utilização. Até a cirurgia, os animais foram alojados em uma instalação de barreira livre de patógenos e desde ração autoclavada e filtrada libdium ad água. Cada gaiola do rato foi fornecido com um ar dedicado e água, e quartos foram mantidos em um ciclo de dia-noite de 12 h. Após a recuperação da cirurgia, os ratos foram mantidos em gaiolas com tops filtrados, desde autoclavado libidum anúncio de comida, e desde que a água filtrada a partir de uma garrafa de água. 1. Preparação de Instrumentos <ol><li> Faça 6 afastadores pele usando clipes de papel e pinos. Torção endireitou clipes de papel nas hastes de papel pinos, lbeiral de um fio de aço resistente cinco centímetros em uma extremidade e fazer uma única 0,5 centímetro &quot;L&quot; em forma de gancho na extremidade do fio. </li><li> Faça cerca de 15 x 15 centímetros campos cirúrgicos quadrados usando filme plástico. Prepare um molho por mouse. Coloque uma torre de papel entre cada envoltório. </li><li> Esterilizar todos os instrumentos cirúrgicos, juntamente com uma pilha de telhas de 12 x 12 polegadas de cortiça, gazes, e cotonetes. </li></ol> 2. Preparação do rato <ol><li> Induzir a anestesia com isoflurano a 2%. Pesar animal. </li><li> Em uma área dedicada preparação cirúrgica raspar tórax esquerdo e pescoço com barbeador elétrico. </li><li> Aplicar uma gota de lágrima artificial do unguento para os olhos do rato. </li><li> Descontaminar pescoço e tórax esquerdo com clorexidina e álcool isopropílico. Repita mais duas vezes. </li></ol> Intubation 3. Rato Oro-traqueal e ventilação mecânica <ol><li> Ter um lugar não-estéril técnico cirúrgico o supino do mouse na área cirúrgica pré-aquecido.</li><li> Confirme profundidade da anestesia, documentando a falta de uma resposta a pata pitada. </li><li> Depois de lavar as mãos e vestindo traje cirúrgico, máscara e chapéu, don luvas cirúrgicas estéreis. </li><li> Depois de drapeados e utilizando uma técnica asséptica, fazer uma incisão vertical um centímetro sobre o mid-neck anterior para expor a laringe. Levemente retrair os músculos da cinta com curvas, serrilhadas 10 centímetros forceps e expor a laringe e traquéia, espalhando os músculos da cinta com a ponta de uma tesoura reta. </li><li> Oralmente inserir um 22 G de ponta romba angiocateter em meados da década de traquéia (Figura 1A) e visualmente confirmar a colocação (Figura 1B). Manter a anestesia e ventilar usando 1-3% de isoflurano através roedor ventilador (225 mL por acidente vascular cerebral; 200 AVC por min). Empregar um limite de pressão de 15 cm H 2 O. </li></ol> 4. Rato Pneumonectomia <ol><li> Coloque o mouse em decúbito lateral direito, com frente para as costas do ratoo operador (lateral-se à esquerda). Usar um filme plástico auto-selante como um campo estéril. Corte através da cortina, usar tesoura curva sem corte para fazer um 2 cm de comprimento corte paralelo às costelas no 4º e 5º espaço intercostal. Insira a ponta curvada tesoura sem corte e dissecar a pele longe das costelas subjacentes e músculos intercostais. </li><li> Retirar a pele com quatro afastadores para expor um quadrado de 1,5 x 1,5 centímetros janela cirúrgica (Figura 2A). Prenda os retratores à placa de cortiça. </li><li> Dissecar até costelas usando uma pinça curva, e usar uma ponta da pinça curvas para entrar na cavidade torácica. </li><li> Usando as micro-tesoura com ponta romba, utilizar a lâmina inferior para entrar na cavidade torácica. Faça um 0,5 centímetros incisão entre as costelas e repita na direção oposta. </li><li> Usando os dois afastadores restantes, abrir o tórax no eixo ântero-posterior e o seguro dos retratores para a placa de cortiça (Figura 2B). </li> &lt;li&gt; Usando pinças curvas de ponta romba na mão esquerda, o pulmão esquerdo agarrar e deslocar a parte superior do pulmão esquerdo lateralmente e inferiormente através da toracotomia até a artéria pulmonar esquerda e brônquios são exposto (Figura 3A, B). </li><li> Segurando o titânio microclipe vascular aplicador carregado na mão direita com o corpo do aplicador na palma e ponta curvada apontando para fora a partir da palma (Figura 3C), deslize a ponta do aplicador para o tórax ao longo da curvatura da face posterior da esquerda pulmão e prender o brônquio esquerdo e artéria pulmonar (Figura 3D). </li><li> Retire o aplicador, mas manter o pulmão esquerdo retraído. Segure as contundentes micro-tesoura de ponta com a mão direita e cortar o brônquio e distal da artéria pulmonar ao clipe e remover pulmão esquerdo (Figura 3E). </li><li> Retire os afastadores de costela. </li><li> Use a pinça sem corte curvas para beliscar até 1 cm de pele inferior ao incisiem, mas acima do nível do diafragma e inserir um angiocateter de 24 L através da pele e para dentro da cavidade torácica esquerda (Figura 4A, B). </li><li> Use 5-0 prolene sutura para colocar dois pontos separados em torno da 4ª e 5 ª costelas para fechar a cavidade torácica. </li><li> Remover os afastadores de pele. Usar dois conjuntos de pinças para aproximar a pele ao longo do comprimento da incisão e cola a pele fechada. </li><li> Conecte-se a 3 ml seringa luer-lock ao angiocateter e remover o ar residual através da aplicação de sucção suave e retirada do angiocateter. </li><li> Cole a incisão no pescoço fechada com dois conjuntos de fórceps, como antes. </li></ol> 5. Rato Sham Pneumonectomia <ol><li> Exponha o pulmão esquerdo como observado no protocolo &quot;Rato Pneumonectomia&quot;. Levantar a caixa torácica com uma pinça sem corte curvas para permitir que o ar na cavidade torácica esquerda (Figura 5A, B). </li><li> Coloque um G angiocateter 24 para o tho esquerdaRacic cavidade como acima tomando cuidado para não ferir o pulmão esquerdo. </li><li> Usando 5-0 prolene sutura e tomando cuidado para não perfurar o pulmão (Figura 5C), coloque dois pedaços de material de sutura na 3ª / 4ª e 5ª / 6 interspaces th costela (Figura 5D). Coloque os dois pedaços de material de sutura antes de amarrar para diminuir os riscos de hérnia de pulmão esquerdo. Amarre o material de sutura para fazer dois pontos separados (Figura 5E). </li><li> Cole a pele sobre a incisão torácica, remover o ar residual com o angiocateter e cole a incisão no pescoço, como acima. </li></ol> 6. Resuscitation, Analgesia, e Recovery <ol><li> Desligue o isoflurano, e administrar 0,1 mg / kg de buprenorfina e 0,5 ml de solução salina normal subcutaneamente. </li><li> Quando respirações espontâneas retomar, remova o tubo endotraqueal. </li><li> Observe rato até que seja novamente ambulatorial. Andando normalmente resumes vários minutos após a remoção do tubo endotraqueal. </li><li> Posicione o mouse em um 27 ° C incubadora (umidificado, 25% de oxigênio) para recuperar O / N. NOTA: Nós colocamos vários pellets de ração umedecida com água no chão da gaiola para as primeiras 24 horas após a cirurgia. </li><li> Administrar 0,1 mg / kg de buprenorfina por via intraperitoneal duas vezes por dia durante três dias após a cirurgia. Tome cuidado para não abrir o local da cirurgia ao manusear animais. </li></ol> 7. Rato Monitoring <ol><li> Pesar camundongos em 1, 3, 5, e 7 dias após a cirurgia. </li></ol>

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The authors have nothing to disclose.

Nós fornecemos a descrição mais detalhada dos procedimentos PNX PNX rato e rato sham reportados à data. Fizemos o leitor ciente de várias das armadilhas comuns que os investigadores de aprendizagem do procedimento comumente encontrar, e nós descrevemos várias técnicas desenvolvidas por nosso laboratório para mitigar contra essas armadilhas. Outros laboratórios que utilizam este modelo pode ter desenvolvido outras modificações na técnica ou utilizar diferentes instrumentos. Ao avaliar as diferenças de técn...

A principal função do pulmão é o de proporcionar para o oxigénio e dióxido de carbono troca entre um organismo e a atmosfera. Nos seres humanos, uma série de condições congênitas e adquiridas levar à redução da área de superfície do pulmão o que resulta em redução da função pulmonar. Apesar de uma série de terapias, como corticóides inalatórios, broncodilatadores, oxigênio suplementar e ventilação mecânica crônica são usados ​​para atenuar as consequências da redução da função pulmonar 1-3, a terapia ideal para essas condições seria promover o crescimento do tecido pulmonar funcional - ou seja, pulmão regeneração. Regeneração de tecidos de mamíferos tem sido bem documentada. O espinhoso Rato Africano pode regenerar grandes áreas da pele, sem a formação de cicatrizes 4. A falange distal em seres humanos podem se regenerar após uma lesão ou amputação 5-7. Seguindo pneumonectomy (PNX), o crescimento compensatório pulmonar ocorre em camundongos 8, 9 ratos, fazergs 10, e os humanos 11. Por definição, o crescimento compensatório pulmonar envolve não apenas a expansão dos espaços aéreos existentes, mas re-septation desses espaços aéreos ampliada com a expansão da microcirculação associada 12. Análise da expressão gênica tem demonstrado que este modelo recapitula muitos dos eventos de sinalização de desenvolvimento pulmonar 13. Quatro semanas após rato PNX, área superficial alveolar é equivalente ao de animais operados simulados 14. Neste artigo, são apresentadas a PNX mouse e procedimentos PNX sham.

<table><tbody><tr><td>6 inch Vascular clip applicator</td><td>Teleflex Medical (WECK)</td><td>137062</td><td></td></tr><tr><td>Horizon small titanium red clip</td><td>Teleflex Medical (WECK)</td><td>1201</td><td></td></tr><tr><td>Narrow pattern 12 cm curved forceps</td><td>Fine Science Tools</td><td>11003-12</td><td></td></tr><tr><td>Curved serrated 10 cm Graefe forceps</td><td>Fine Science Tools</td><td>11052-10</td><td></td></tr><tr><td>Castroviejo needle holder</td><td>Fine Science Tools</td><td>12565-14</td><td></td></tr><tr><td>Straight 9 cm Strabismus scissors (blunt tip)</td><td>Fine Science Tools</td><td>14075-09</td><td></td></tr><tr><td>Straight 8.5 cm hardened fine scissors</td><td>Fine Science Tools</td><td>14090-09</td><td></td></tr><tr><td>Straight, blunt tip Cohan-Vannas spring scissors</td><td>Fine Science Tools</td><td>15000-12</td><td></td></tr><tr><td>Skin glue</td><td>Gluture</td><td>32046</td><td></td></tr><tr><td>22 G Angiocatheter</td><td></td><td></td><td></td></tr><tr><td>24 G Angiocatheter</td><td></td><td></td><td></td></tr><tr><td>3 ml Luer lock syringe</td><td></td><td></td><td></td></tr><tr><td>4 Short retractors</td><td></td><td></td><td></td></tr><tr><td>2 Long retractors</td><td></td><td></td><td></td></tr><tr><td>5-0 Prolene on curved cutting needle</td><td>Ethicon</td><td>8698G</td><td></td></tr><tr><td>0.5 ml Syringe on 27 G needle</td><td></td><td></td><td></td></tr><tr><td>Normal saline</td><td></td><td></td><td></td></tr><tr><td>Buprenorphine</td><td></td><td></td><td></td></tr><tr><td>Press-n-Seal wrap</td><td>Glad Products Company</td><td></td><td></td></tr><tr><td>12 x 12 inch Cork board stack</td><td>Office Depot</td><td></td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td></td></tr><tr><td>Betadine</td><td></td><td></td><td></td></tr><tr><td>Mouse ventilator</td><td>Hugo Sachs Elektronnik</td><td>Minivent Type 845</td><td></td></tr><tr><td>Isoflurane vaporizer</td><td>OHMEDA</td><td>Excel 210 SE</td><td></td></tr><tr><td>Artificial tear ointment</td><td>Puralube</td><td>NDC: 17033-211-38</td><td></td></tr></tbody></table>

mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

. Um lote de PNX e operação simulada rato pesos é fornecido na Figura 6 Em nossas mãos, a sobrevivência é constantemente 95-100% para ambos PNX e sham pneumonectomy. 14 Para obter descrições de como o pulmão direito re-cresce neste modelo e o curso do tempo esperado, nos referimos ao leitor manuscritos de Gibney 15 et al. E Wang et al. Várias armadilhas comuns devem ser evitados para realizar com êxito os procedimentos sham ...

Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

Mouse Pneumonectomy Model of Compensatory Lung Growth

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Rato Pneumonectomia modelo de crescimento compensatório pulmonar

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.8K visualizações.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Vídeo: Mouse Pneumonectomy Model of Compensatory Lung Growth

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

Desenvolvimento de bronquiolite obliterante em um modelo murino de transplante de pulmão ortotópico

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

Medicina

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

Internal lung surface area (ISA) is a critical criterion for assessing lung morphology and physiology in lung diseases and injury-induced alveolar regeneration. We describe here a standardized method that can minimize the measurement bias for ISA in both lung pneumonectomy and prosthesis implantation mouse models.

Um Método Padronizado para Medição da Superfície de Pulmão Interna via Neumonectomia de Mouse e Implante de Prótese

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

Biologia do Desenvolvimento

Measurement of the Pressure-volume Curve in Mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential phenotypic changes are best assessed by measurement of the changes in lung elasticity. To best understand specific mechanisms underlying such pathologies in mice, it is essential to make functional measurements that can reflect the developing pathology. Although there are many ways to measure elasticity, the classical method is that of the total lung pressure-volume (PV) curve done over the whole range of lung volumes. This measurement has been made on adult lungs from nearly all mammalian species dating back almost 100 years, and such PV curves also played a major role in the discovery and understanding of the function of pulmonary surfactant in fetal lung development. Unfortunately, such total PV curves have not been widely reported in the mouse, despite the fact that they can provide useful information on the macroscopic effects of structural changes in the lung. Although partial PV curves measuring just the changes in lung volume are sometimes reported, without a measure of absolute volume, the nonlinear nature of the total PV curve makes these partial ones very difficult to interpret. In the present study, we describe a standardized way to measure the total PV curve. We have then tested the ability of these curves to detect changes in mouse lung structure in two common lung pathologies, emphysema and fibrosis. Results showed significant changes in several variables consistent with expected structural changes with these pathologies. This measurement of the lung PV curve in mice thus provides a straightforward means to monitor the progression of the pathophysiologic changes over time and the potential effect of therapeutic procedures.

Here we present a protocol to simply and reliably measure the lung pressure-volume curve in mice, showing that it is sufficiently sensitive to detect phenotypic parenchymal changes in two common lung pathologies, pulmonary fibrosis and emphysema. This metric provides a means to quantify the lung&#8217;s structural changes with developing pathology.

Medição da curva pressão-volume no mouse Lungs

In recent decades the mouse has become the primary animal model of a variety of lung diseases. In models of emphysema or fibrosis, the essential ...

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high incidence in aging populations. The current conventional therapies for COPD focus mainly on symptom-modifying drugs; thus, the development of new therapies is urgently needed. Qualified animal models of COPD could help to characterize the underlying mechanisms and can be used for new drug screening. Current COPD models, such as lipopolysaccharide (LPS) or the porcine pancreatic elastase (PPE)-induced emphysema model, generate COPD-like lesions in the lungs and airways but do not otherwise resemble the pathogenesis of human COPD. A cigarette smoke (CS)-induced model remains one of the most popular because it not only simulates COPD-like lesions in the respiratory system, but it is also based on one of the main hazardous materials that causes COPD in humans. However, the time-consuming and labor-intensive aspects of the CS-induced model dramatically limit its application in new drug screening. In this study, we successfully generated a new COPD model by exposing mice to high levels of ozone. This model demonstrated the following: 1) decreased forced expiratory volume 25, 50, and 75/forced vital capacity (FEV25/FVC, FEV50/FVC, and FEV75/FVC), indicating the deterioration of lung function; 2) enlarged lung alveoli, with lung parenchymal destruction; 3) reduced fatigue time and distance; and 4) increased inflammation. Taken together, these data demonstrate that the ozone exposure (OE) model is a reliable animal model that is similar to humans because ozone overexposure is one of the etiological factors of COPD. Additionally, it only took 6 - 8 weeks, based on our previous work, to create an OE model, whereas it requires 3 - 12 months to induce the cigarette smoke model, indicating that the OE model might be a good choice for COPD research.

This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone.

Geração de um modelo de doença pulmonar obstrutiva crônica em ratos pela exposição repetida de ozônio

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

The Left Pneumonectomy Combined with Monocrotaline or Sugen as a Model of Pulmonary Hypertension in Rats

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats with the additional administration of monocrotaline (MCT) or SU5416 (Sugen). We also compare these two models to other PAH models commonly used in research. In the last few years, the focus of animal PAH models has moved towards studying the mechanism of angioproliferation of plexiform lesions, in which the role of increased pulmonary blood flow is considered as an important trigger in the development of severe pulmonary vascular remodeling. One of the most promising rodent models of increased pulmonary flow is the unilateral left pneumonectomy combined with a "second hit" of MCT or Sugen. The removal of the left lung leads to increased and turbulent pulmonary blood flow and vascular remodeling. Currently, there is no detailed procedure of the pneumonectomy surgery in rats. This article details a step-by-step protocol of the pneumonectomy surgical procedure and post-operative care in male Sprague-Dawley rats. Briefly, the animal is anesthetized and the chest is opened. Once the left pulmonary artery, pulmonary vein, and bronchus are visualized, they are ligated and the left lung is removed. The chest then closed and the animal recovered. Blood is forced to circulate only on the right lung. This increased vascular pressure leads to a progressive remodeling and occlusion of small pulmonary arteries. The second hit of MCT or Sugen is used one week post-surgery to induce endothelial dysfunction. The combination of increased blood flow in the lung and endothelial dysfunction produces severe PAH. The primary limitation of this procedure is that it requires general surgical skills.

The rodent left pneumonectomy is a valuable technique in pulmonary hypertension research. Here, we present a protocol to describe the rat pneumonectomy procedure and postoperative care to ensure minimal morbidity and mortality.

A pneumonectomia esquerda combinado com Monocrotaline ou Sugen como modelo de hipertensão pulmonar em ratos

In this protocol, we detail the correct procedural steps and necessary precautions to successfully perform a left pneumonectomy and induce PAH in rats ...

Experimental Model to Evaluate Resolution of Pneumonia

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to high morbidity and mortality. Currently, there are no pre-clinical models that recapitulate the complexity of human ARDS. However, infectious models of pneumonia (PNA) can replicate the main pathophysiological features of ARDS. Here, we describe a model of PNA induced by the intratracheal instillation of live Streptococcus pneumoniae and Klebsiella pneumoniae in C57BL6 mice. In order to evaluate and characterize the model, after inducing injury, we carried out serial measurements of body weight and bronchoalveolar lavage (BAL) for measuring markers of lung injury. Additionally, we harvested lungs for cell count and differentials, BAL protein quantification, cytospin, bacterial colony-forming unit counts, and histology. Lastly, high dimensional flow cytometry was performed. We propose this model as a tool to understand the immune landscape during the early and late resolution phases of lung injury.

This manuscript describes the establishment of an infectious model of pneumonia in mice and the respective characterization of injury resolution along with methods for growing bacteria and intratracheal instillation. A novel approach using high-dimensional flow cytometry to evaluate the immune landscape is also described.

Modelo experimental para avaliar a resolução da pneumonia

Acute respiratory distress syndrome (ARDS) causes acute lung injury, characterized by rapid alveolar damage and severe hypoxemia. This, in turn, leads to ...

Imunologia e Infecção

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.

Modelo de Lesão de Isquemia/Reperfusão de Pulmão de Rato em Transplante de Pulmão de Rato: Otimizações para Melhorar os Resultados

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

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ quality exacerbated by excessive geographic transportation distance and stringent donor organ acceptance criteria pose limitations to current LTx efforts. Ex situ lung perfusion (ESLP) is an innovative technology that has shown promise in attenuating these limitations. The physiologic ventilation and perfusion of the lungs outside of the inflammatory milieu of the donor body affords ESLP several advantages over traditional cold static preservation (CSP). There is evidence that negative pressure ventilation (NPV) ESLP is superior to positive pressure ventilation (PPV) ESLP, with PPV inducing more significant ventilator-induced lung injury, pro-inflammatory cytokine production, pulmonary edema, and bullae formation. The NPV advantage is perhaps due to the homogenous distribution of intrathoracic pressure across the entire lung surface. The clinical safety and feasibility of a custom NPV-ESLP device have been demonstrated in a recent clinical trial involving extender criteria donor (ECD) human lungs. Herein, the use of this custom device is described in a juvenile porcine model of normothermic NPV-ESLP over a 12 h duration, paying particular attention to management techniques. Pre-surgical preparation, including ESLP software initialization, priming, and de-airing of the ESLP circuit, and the addition of anti-thrombotic, anti-microbial, and anti-inflammatory agents, is specified. The intraoperative techniques of central line insertion, lung biopsy, exsanguination, blood collection, cardiectomy, and pneumonectomy are described. Furthermore, particular focus is paid to anesthetic considerations, with anesthesia induction, maintenance, and dynamic modifications outlined. The protocol also specifies the custom device's initialization, maintenance, and termination of perfusion and ventilation. Dynamic organ management techniques, including alterations in ventilation and metabolic parameters to optimize organ function, are thoroughly described. Finally, the physiological and metabolic assessment of lung function is characterized and depicted in the representative results.

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

This paper describes a porcine model of negative pressure ventilation ex situ lung perfusion, including procurement, attachment, and management on the custom-made platform. Focus is made on anesthetic and surgical techniques, as well as troubleshooting.

Perfusão pulmonar ex situ com pressão negativa normotérmica: avaliação da função e metabolismo pulmonar

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

A Simple Cuff Technique for Murine Left Lung Transplantation

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an efficient and highly reliable "cuff technique" of transplantation. This article describes a sophisticated and comprehensive method for orthotopic lung transplantation in a vascularized orthotopic lung model, representing the most physiologic and clinically relevant model of mouse lung transplantation to date. The transplantation process consists of two distinct stages: donor harvest and subsequent implantation into the recipient. The method has been successfully mastered, and with several months of sufficient training, a skilled practitioner can perform the procedure in approximately 90 min from skin-to-skin. Surprisingly, once individuals overcome the initial learning curve, the survival rate during the perioperative period approaches nearly 100%. The mouse model allows for the use of multiple commercially available transgenic and mutant strains of mice, enabling the study of tolerance and rejection. Additionally, the unique features of this model make it a valuable tool for investigating tumor biology and immunology.

<meta charset="utf8"></head><body>Uma técnica simples de manguito para transplante de pulmão esquerdo murino

The present protocol describes a cuff technique for a mouse left lung transplantation model. This technique has been developed over several years and has performed well, serving effectively in immunological research.

Uma técnica simples de manguito para transplante de pulmão esquerdo murino

Over the past decade, our laboratory has made significant progress in developing and refining vascularized mouse lung transplantation models using an ...

Rato Pneumonectomia modelo de crescimento compensatório pulmonar

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Medição da curva pressão-volume no mouse Lungs

Geração de um modelo de doença pulmonar obstrutiva crônica em ratos pela exposição repetida de ozônio

A pneumonectomia esquerda combinado com Monocrotaline ou Sugen como modelo de hipertensão pulmonar em ratos

Modelo experimental para avaliar a resolução da pneumonia

Modelo de Lesão de Isquemia/Reperfusão de Pulmão de Rato em Transplante de Pulmão de Rato: Otimizações para Melhorar os Resultados

Perfusão pulmonar ex situ com pressão negativa normotérmica: avaliação da função e metabolismo pulmonar

Uma técnica simples de manguito para transplante de pulmão esquerdo murino

Uma técnica simples de manguito para transplante de pulmão esquerdo murino