Name: mouse pneumonectomy model of compensatory lung growth
Uploaded: 2014-12-17T14:00:00.000+00:00
Duration: 9 min 22 s
Description: Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

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.

NOT: Hayvan kullanım bildirimi: Bu çalışmada tüm işlemler onay ve Cincinnati Çocuk Hastanesi Kurumsal Hayvan Kullanımı ve Bakımı Komitesi (IACUC) yönergeleri takip gerçekleştirilmiştir. Sekiz hafta yaşında C57BL / 6J erkek fareleri, Jackson Laboratories&#39;den (Bar Harbor, ME) elde edilir ve kullanımdan önce bir hafta boyunca ortama alışmaları sağlandı. Cerrahi kadar, hayvanlar patojen madde içermeyen bir yere yerleştirilmesi ve otoklavlanmıştır yemi ve süzüldü suyu ad libdium sağlandı. Her fare kafesi özel hava ve su ile sağlanan ve odalar 12 saat gece-gündüz döngüsü tutuldu. Cerrahi işlemden sonra, fareler, süzüldü üstleri ile kafeslerde muhafaza otoklavlanmış yemi farelere istenildiği kadar mesafede, ve bir su şişesinden su süzüldü sağlandı. Araçların 1. Hazırlık <ol><li> Ataç ve işaretçilerine kullanılarak 6 cilt Ekartörleri olun. Büküm kağıt pim Shanks üzerine ataç doğruldu, lbir ucunda bir 5 cm&#39;lik tek bir çelik tel eave ve telin ucunda tek bir 0.5 cm&#39;lik bir &quot;U&quot; şekilli bir kancayı olun. </li><li> Plastik wrap kullanarak bazı 15 x 15 cm kare cerrahi perdeler olun. Fare başına bir pansuman hazırlayın. Her şal arasında bir kağıt kule koyun. </li><li> 12 x 12 inç mantar fayans, gazlı bez ve pamuklu çubuklar bir yığını ile birlikte tüm cerrahi aletler sterilize. </li></ol> 2. Fare Hazırlama <ol><li> % 2 izofluran ile anestezi neden olur. Hayvan tartılır. </li><li> Özel cerrahi hazırlık alanında elektrik tıraş sol toraks ve boyun alanı tıraş. </li><li> Farenin gözlerine yapay gözyaşı merhemi bir damla uygulayın. </li><li> Klorheksidin ve izopropil alkol ile boyun ve sol toraks arındırın. Iki kez daha tekrarlayın. </li></ol> 3. Fare Oro-trakeal entübasyon ve Mekanik Ventilasyon <ol><li> Steril olmayan bir cerrahi teknisyen yerde önceden ısıtılmış cerrahi alanında fare sırtüstü var.</li><li> Pençe tutam yanıt eksikliği belgeleyerek anestezi derinliği onaylayın. </li><li> Cerrahi kıyafetleri, maske, şapka ve ellerini yıkama ve donning sonra, steril cerrahi eldiven yok. </li><li> Dökümlü ve aseptik teknik kullanılarak sonra, gırtlak maruz ön orta boyun üzerinde 1 cm&#39;lik dikey kesi yapmak. Hafif kavisli, tırtıklı 10 cm forseps ile kayış kasları geri çekin ve düz makas ucu ile kayış kasları yayarak gırtlak ve trakea maruz. </li><li> Oral orta trakea (Şekil 1A) içine 22 G künt uçlu angiocatheter yerleştirin ve görsel yerleştirme (Şekil 1B) onaylayın. Anestezi koruyun ve kemirgen ventilatör ile (inme başına 225 ul; min başına 200 Stokes)% 1-3 izofluran kullanarak havalandırın. 15 cm H2O bir basınç sınırını ayrılmadan </li></ol> 4. Fare Pnömonektomi <ol><li> Farenin arka bakacak sağ lateral dekübit pozisyonunda fare LayOperatör (sol taraf yukarı). Steril örtü olarak kendi kendine kapanan plastik wrap kullanın. Drape ile kesme, 4. ve 5. interkostal aralıktan kaburga 2 cm uzunluğunda kesilmiş paralel hale getirmek için künt uçlu kavisli makas kullanın. Kör uç kavisli makas yerleştirin ve alttaki kaburga ve interkostal kasların uzak cildi teşrih. </li><li> Bir kare 1.5 x 1.5 cm cerrahi pencere (Şekil 2A) maruz dört toplayıcıları ile cildi geri çekin. Mantar kuruluna Ekartörleri sabitleyin. </li><li> Kavisli bir forseps kullanarak kaburga aşağı incelemek ve göğüs boşluğu girmek için kavisli bir forseps bir ucunu kullanın. </li><li> Künt ucu mikro-makas kullanarak, göğüs boşluğu girmek için alt bıçak kullanın. Ters yönde kaburga ve tekrarı arasında 0,5 cm&#39;lik kesi yapmak. </li><li> Kalan iki Ekartörleri kullanarak, ön-arka eksen ve mantar pano (Şekil 2B) Retraktörler güvenli olarak toraks açın. </li> &lt;Sol pulmoner arter ve bronş (Şekil 3A, B) maruz kadar sol elinde kavisli künt uçlu forseps kullanarak li&gt; torakotomi ile yanal ve inferior sol akciğer üst kısmını sol akciğer kavramak ve yerinden. </li><li> Avuç aplikatör ve avuç (Şekil 3C) uzak işaret kavisli ucu gövdesi ile sağ elinde yüklü titanyum vasküler mikroklip aplikatör Holding, sol arka yönü eğrilik boyunca toraks içine aplikatör ucu kaydırın Akciğer ve bronşları sol ve pulmoner arter (Şekil 3D) klip. </li><li> Aplikatör çıkarın ama geri sol akciğer tutun. Sağ eli ile künt ucu mikro-makas kavrayın ve klibe bronşları ve pulmoner arter distalinde kesilir ve sol akciğer (Şekil 3E) çıkarın. </li><li> Kaburga Ekartörleri çıkarın. </li><li> İncisi aşağı cilt 1 cm kadar çimdik kavisli künt forseps kullanındeğil, diyafram seviyesinden ve deri yoluyla ve sol göğüs boşluğunda (Şekil 4A, B) 24 G angiocatheter yerleştirin. </li><li> Göğüs boşluğu kapatmak için 4 th yaklaşık iki kesintiye sütür ve 5 inci kaburga yerleştirmek için 5-0 prolen sütür kullanın. </li><li> Cilt Ekartörleri çıkarın. Kesi uzunluğu boyunca cildi yaklaştığı ve cilt kapalı tutkal forseps iki takım kullanın. </li><li> Angiocatheter 3 ml&#39;lik luer-lock şırınga bağlayın ve nazik vakum uygulayarak ve angiocatheter çekerek artık havayı çıkartın. </li><li> Boyun kesi daha önce olduğu gibi forseps iki takım kullanılarak kapalı Tutkal. </li></ol> 5. Fare Sham Pnömonektomi <ol><li> &quot;Fare Pnömonektomi&quot; protokolde belirtildiği gibi sol akciğer Açığa. Sol göğüs boşluğuna hava girmesini sağlamak için kavisli künt forseps ile göğüs kafesini kaldırın (Şekil 5A, B). </li><li> Sol tho içine 24 G angiocatheter yerleştirinSol akciğerin kesmemeye özen yukarıdaki gibi Toraks boşluğu. </li><li> 5-0 prolen sütür kullanarak ve akciğer (Şekil 5C) delinme dikkatli olmak, 3. / 4 th içine dikiş materyali iki uzunlukları ve 5. / 6. kaburga aralar (Şekil 5D) yerleştirin. Sol akciğer herniasyonu risklerini azaltmak için ipe önce dikiş materyali her iki uzunlukları yerleştirin. İki kesintiye dikiş (Şekil 5E) yapmak için sütür materyali bağlayın. </li><li> Torasik kesi üzerindeki deriyi Tutkal, angiocatheter ile artık havayı kaldırmak ve yukarıdaki gibi boyun kesi tutkal. </li></ol> 6. Resüsitasyon, Analjezi ve Kurtarma <ol><li> Izofluran kapatın ve buprenorfın 0.1 mg / kg yönetmek ve serum fizyolojik subkutan 0.5 ml. </li><li> Spontan solunum frekanslarında devam zaman, endotrakeal tüp kaldırmak. </li><li> Tekrar ayaktan kadar fareyi gözlemleyin. Tipik resum YürüyüşEndotrakeal tüp çıkarılmasından sonra birkaç dakika es. </li><li> / N O kurtarmak için bir 27 ° C inkübatör (nemlendirilmiş,% 25 oksijen) fare yerleştirin. NOT: Biz ameliyat sonrası ilk 24 saat için kafes katta su ile ıslatılmış yem birkaç topakları yerleştirin. </li><li> Ameliyat sonrası, günde iki kez, üç gün boyunca intraperitonal enjeksiyon ile buprenorfin, 0.1 mg / kg uygulayın. Hayvanları tutarken cerrahi siteyi açmak için özen gösterin. </li></ol> 7. Fare İzleme <ol><li> Ameliyat sonrası 1, 3, 5 ve 7 gün sonra fareler tartılır. </li></ol>

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The authors have nothing to disclose.

Biz bugüne kadar bildirilen fare PNX ve fare sahte PNX prosedürleri en detaylı açıklamasını sağladı. Biz prosedürü öğrenme araştırmacılar sık ​​karşılaştıkları ortak tuzaklar birkaç okuyucu farkında yaptık, ve biz bu tuzaklar karşısında azaltmak için laboratuvar tarafından geliştirilen çeşitli teknikler belirttiğimiz. Bu modeli kullanan diğer laboratuvarlar diğer bir teknik değişiklikler geliştirilen veya farklı enstrümanlar kullanmak olabilir. Teknikleri farklılıkları de?...

akciğerin başlıca fonksiyonu, bir organizmaya ve atmosfer arasındaki oksijen ve karbon dioksit değişimi sağlamaktır. İnsanlarda, doğumsal ve edinsel koşulları bir dizi bozulmuş akciğer fonksiyonu ile sonuçlanan azalmış akciğer yüzey alanına yol açar. Yani, akciğer - örneğin inhale kortikosteroidler, bronkodilatör, ek oksijen, ve kronik mekanik ventilasyon gibi tedavilerin bir ev sahibi engelli akciğer fonksiyonu 1-3 sonuçlarını hafifletmek için kullanılmasına rağmen, bu koşullar için ideal bir tedavi fonksiyonel akciğer dokusunun yeniden büyümesini teşvik ediyorum yenilenmesi. Memeli doku rejenerasyonu iyi belgelenmiştir. Afrika Dikenli Fare skar oluşumu 4 olmaksızın cildin geniş alanlar yeniden yapabilirsiniz. İnsanlarda distal falanks yaralanma veya ampütasyon 5-7 aşağıdaki yeniden yapabilirsiniz. Aşağıdaki pnömonektomi (PNX), telafi akciğer büyümesi yapmak, 9 fare, sıçan 8 oluşurgs 10, ve 11 insanlarda. Tanım olarak, telafi akciğer büyümesi ilişkili mikrosirkülasyon 12 genişlemesi ile bu genişlemiş hava sahasındaki mevcut hava sahasındaki değil sadece genişleme, ancak yeniden bölümlenmenin içerir. Gen ifadesi analizi bu modeli akciğer gelişimi 13 sinyalizasyon olaylar birçok özetlediği olduğunu göstermiştir. Dört hafta, fare PNX sonra, alveol yüzey alanının ve sahte işlemden geçirilmiş hayvan 14 edilene eşdeğerdir. Bu yazıda, fare pNX ve sahte PNX prosedürlerini açıklar.

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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.

. Hem PNX ve sahte pnömonektomi için% 100 - pNX ve Sham fare ağırlıkları bir arsa Şekil 6&#39;da verilmiştir Bizim ellerde, hayatta sürekli 95. Sağ akciğer-yetişir yeniden bu model ve beklenen zaman ders, biz Gibney ark. 15 ve Wang ve ark yazmaları okuyucuyu bakın nasıl açıklamaları için. 14 Birkaç ortak tuzaklar başarıyla fare PNX ve fare sahte pnömonektomi prosedürleri gerçekleştirmek için kaçın?...

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.

Telafi Akciğer Büyüme Fare Pnömonektomi Modeli

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

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12.8K Görüntüleme.  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.

Video: 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.

Ortotopik Akciğer Transplantasyonu bir Mürin Modelinde Obliteratif Bronşiolit Gelişimi

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

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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.

Fare Pnömonektomi ve Protez İmplantasyonu ile İç Akciğer Yüzey Alanının Ölçümü için Standart Bir Yöntem

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

Gelişim Biyolojisi

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.

Fare Akciğerler Basınç-hacim Eğrisi Ölçümü

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.

Kronik obstrüktif akciğer hastalığı manken tekrarlanan ozon maruz farelerde nesil

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.

Sıçanlarda pulmoner hipertansiyon bir Model olarak Monocrotaline ya da Sügen ile birlikte sol Pneumonectomy

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.

Pnömoni Çözünürlüğünü Değerlendirmek için Deneysel Model

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

İmmünoloji ve Enfeksiyon

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.

Sıcak iskemi / reperfüzyon hasarının bir sıçan akciğer transplantasyon modeli: sonuçları iyileştirmek için optimizasyonlar

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.

Norannemik Negatif Basınçlı Ventilasyon Ex Situ Akciğer Perfüzyonu: Akciğer Fonksiyonlarının ve Metabolizmasının Değerlendirilmesi

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.

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.

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

Ex Vivo Porcine Experimental Model for Studying and Teaching Lung Mechanics

Mechanical ventilation is widely used and requires specific knowledge for understanding and management. Health professionals in this field may feel insecure and lack knowledge because of inadequate training and teaching methods. Therefore, the objective of this article is to outline the steps involved in generating an ex vivo porcine lung model to be used in the future, to study and teach lung mechanics. To generate the model, five porcine lungs were carefully removed from the thorax following the guidelines of the Animal Research Ethics Committee with adequate care and were connected to the mechanical ventilator through a tracheal cannula. These lungs were then subjected to the alveolar recruitment maneuver. Respiratory mechanics parameters were recorded, and video cameras were used to obtain videos of the lungs during this process. This process was repeated for five consecutive days. When not used, the lungs were kept refrigerated. The model showed different lung mechanics after the alveolar recruitment maneuver every day; not being influenced by the days, only by the maneuver. Therefore, we conclude that the ex vivo lung model can provide a better understanding of lung mechanics and its effects, and even of the alveolar recruitment maneuver through visual feedback during all stages of the process.

Ex Vivo Porcine Experimental Model for Studying and Teaching Lung Mechanics

We present an ex vivo pig lung model for the demonstration of pulmonary mechanics and alveolar recruitment maneuvers for teaching purposes. The lungs can be used for more than one day (up to five days) with minimal changes in pulmonary mechanics variables.

Ex Vivo (Canlı Canlı) Akciğer Mekaniğini İncelemek ve Öğretmek için Domuz Deneysel Modeli

Mechanical ventilation is widely used and requires specific knowledge for understanding and management. Health professionals in this field may feel ...