Biz akciğer döküm ile yardımcı Dr Adlah Sukkar, MD işi kabul ederler. Bu çalışma NHLBI, HL088005 tarafından finanse edilmiştir.

Sıçanlar üzerinde gerçekleştirilen tüm protokoller Johns Hopkins Üniversitesi Hayvan Bakım ve Kullanım Kurulu tarafından ve NIH yönergeleri doğrultusunda onaylanmıştır. Mümkün olduğunda, hayvan cerrahi ameliyat kontaminasyon en aza indirmek için cerrahi bölgeden ayrı bir alanda prepped olmalıdır. 1. Anestezi / analjezi <ol><li> % 3 izofluran ile aşılanmış bir indüksiyon odasında; Yeri sıçan (Harlan, Indianapolis, IN Sprague Dawley erkek rat, 125-150 g). </li><li> % 3 izofluran anestezi ile burun konisi ve ventilatöre bağlı cerrahi gemide anestezi sıçan yerleştirin. Tüm prosedürler için steril tekniği kullanın. Steril çalışma alanı sağlamak için fare asmak. </li></ol> 2. Entübasyon <ol><li> Cerrahi bant kullanarak yatar pozisyonda uzantıları hareketsiz. </li><li> Burun konisi, sıçan kaldır, yastıklı forseps ile dil uzatmak. </li><li> Kılavuz olarak küntleşmiş metal Stilet ile 14 gauge intracath kullanın. Geride kaydırıntrakea içine ngue. </li><li> Trakeada beyaz plastik intracath bırakarak metal Stilet çıkarın. Sıçan nefes ve hava borusu akan emin olun. </li><li> Kateter doğrudan adaptör ile burun konisi değiştirin, ventilatör (; 8 ml / kg tidal volüm; Kemirgen Vantilatör Model 683, Harvard Apparatus, Holliston, Massachusetts 90 solunum / dk) sıçan bağlayın. </li><li> Puralube (Butler Schein, Dublin, OH) gözleri Veteriner merhem uygulayın. </li></ol> 3. Torakotomi <ol><li> Sıra sıçan sağ tarafında aşağı, cerrahi bant kullanarak uzantıları hareketsiz. </li><li> Sol tarafında göğüs kafesi alanı Tıraş. </li><li> Küçük vakum ile aşırı kürk çıkarın. </li><li> Povidon-İyot swabstick (Dynarex Corporation Orangebur, NY) ardından alkol ile silinerek steril alanın olun. Bu işlem iki kez daha (üç scrubs toplam) tekrarlayın. </li><li> Alanın merkezinde neşter steril kesme makası veya steril ile transvers kesi olun. </li><li> L boyunca teşrih küntaşağı kaburga (son kat kaburga kaplayan ince bir zardır) doku ve yağ ayers. </li><li> 3. interkostal alan belirlemek için kaburga sayısı. </li><li> Steril 45 ° Graefe forseps kullanarak, 3. ve 4. kaburga arasında künt kesi yapmak. </li><li> , Kaburga ayırıcılar yerleştirin hafifçe açık tutmak için sütürler açık akciğer tam görselleştirme void yer bandı oluşturma çekin. </li></ol> 4. Sol Pulmoner Arter Ligasyonu <ol><li> 90 ° Graefe forseps ile, sağ el ile sol arka akciğer taşıyın. </li><li> Dumont pattern # 5 düz forseps kullanarak, sol eliyle sol pulmoner arter ve havayolu kapmak. Sol pulmoner arter yolu üstünde yatıyordu olacaktır. Forseps tabloya doğrudan dik olduğundan emin olun. Ayrıca, en distal pozisyonlar (parankim yakın) de sol pulmoner arter / sol ana bronşu almak için en etkili yöntemdir. Bu sol manevra ile kenara havalandırma sol akciğer itin. </li><li> Dumont kullanınpattern # 5 45 ° kendi doğal sınırları, sol ana bronşu gelen sol pulmoner arter ayırmak için kavisli bir forseps. Bu ayırma çizgisi iki ayrı yapı arasındaki ince ve beyaz görünür. </li><li> Damar geçmeden arter altında doğrudan ayırın; sorunsuz ayrılması boyunca birlikte düzenlenen forseps ipuçları kaydırın. </li><li> Forseps ipuçları gözle sol pulmoner arter ve sol ana bronşu ayrılır kadar devam edin. Ucu sadece küçük bir nokta tam yoluyla olmalıdır. Kan forseps ucu üzerinde görselleştirilebilir edilebilir ise, o zaman yoluyla tam değildir ve ligasyon teşebbüs edilmemelidir. Bir kez uzayda, sol pulmoner arter forseps eğrisi yatıyordu olacaktır. Bu pozisyonda tutun. </li><li> Yavaşça künt forseps kavrama (sol) bırakın ve bir pre-cut sütür (~ 2-3 inç; polipropilen sütür boyutu 6-0; Myco Tıp, Cary, NC) parça kapmak. </li><li> Sol pulmoner arter ve g kucağında kavisli bir forseps açınrab sütür. Yavaşça forseps eğriye göre yukarıya doğru bir hareketle sol pulmoner arter ve sol ana bronşu arasındaki uzayda sütür çekin. </li><li> Bir kare düğüm ile sol pulmoner, sütürün dikkatle kırt kalanı tıkayıp sıkılaştırın. </li><li> (Mavi monofilaman) polipropilen ile iki kez kaburga ve sütür tutun künt forseps kullanarak Kapat kaburga boyutu dikkatli olmanın, bir 19 mm, bir hemostat iğne (Myco Tıp, Cary, NC) kesim 3/8 daire ters bağlanmış 4-0 sütür cilt (sadece kaburga). </li><li> Gevşek bir kare düğüm tamamlayın, pozitif ekspirasyon sonu basınç (PEEP; 2-5 cmH2O) üzerinde yer akciğer şişirmek, sıkı akciğer ardından güvenli düğüm hyperinflate ve sütür kalanı kırpma önce başka bir tam düğüm yapmak. PEEP çıkar ve akciğeri olmadığından emin olmak için 30 saniye görselleştirmek. 5 damla bupivicaine (APP Pharmaceuticals, Schaumbur, IL) uygulayın. </li><li> Kapat yara üzerine doku yapıştırıcısı yerleştirerek cilt ve birlikte cilt itin tr back kullanarakpamuk uçlu aplikatör d. Daha sonra yara yerinden 24 saat süre ile her 8 saatte ya da hayvan normal aktivite kadar devam eder bupivakain (2.0 mg / kg deri altı) ver. Bu alanda subkutan tabakası çok ince olduğundan, kendi kendine dikiş kolay değildir. Doku yapıştırıcı uygulanır ve deri birlikte itilir, bu da deri altı tabaka kapatır. </li><li> Izofluran gazı kapatın ama istemli hareket dönünceye kadar oda havası 1-2 dakika sıçan havalandırmak için devam ediyor. Ventilatörden trakeal tüp ayırın ve sıçan çıkarmadan önce spontan soluyan olduğundan emin olun. </li><li> Pamuklu çubukla ve monitör hareketi ve kurtarma ile gözlerden Puralube siliniz. (0.05 mg / kg intraperitoneal, Butler Schein, Dublin OH) Buprenorfin hidroklorür enjekte. Ameliyat sonrası 48 saat süreyle analjezik her 12 saatte teslim devam edin. </li></ol> 5. Sol Karotis Arter Kanülasyonu Bronş perfus büyüklüğünü değerlendirmek için,sol pulmoner arter ligasyonu sonrası istenilen zaman noktalarında iskemik sol akciğer iyon, arkus içine sol karotis arter yoluyla etiketli mikrosferler enjekte. 1-2 yukarıdaki gibi sıçan hazırlayın. <ol><li> Boyun boyunca orta hat Kes, trakea ve sol karotis arter (mikroküre enjeksiyon yerinde) ortaya çıkarmak için teşrih künt. </li><li> 25 g iğne, 4 yollu musluk, 1 ml şırınga bağlı damar içine PE20 boru (Becton Dickinson, MD Sparks) ve heparinize salin, künt ucu ile dolu kateter yerleştirin. </li><li> Su sonikatör 30 sn için; mikroküreler (Invitrogen, Eugene, OR x 10 6 küre / ml 15 mikron kızıl polistiren floresan mikrosferler, 1) Yeri şişe. </li><li> Şişe, vorteks çıkarın ve 20 g iğne yoluyla 1 ml Hamilton cam şırınga (Hamilton Company, Reno, NV) içine 0.5 ml (500.000 mikroküreler) hazırlar. </li><li> 4-yollu musluk Hamilton şırınga takılır ve enjektör pompası (oranıyla mikroküreler demlenmeye: 500 ul / dak; Genie Plus, Kent Bilimsel, Torringt) BT üzerinde. </li><li> 500 ul / dk &#39;heparized tuzlu 1 ml Hamilton şırıngası ile aynı hizada cihazları çıkarın. </li><li> Tam göğüs torakotomi gerçekleştirin ve inferior vena kava kopararak sıçan exsanguinate. </li><li> Sol akciğerde ve diğer ilgi dokuları çıkartın. </li><li> Dokudan mikroküreler ayıklamak için, kan kaybından sonra sıçan sol akciğerinin tamamı almak ve 2M KOH (4-6 ml) koyun. 55 ° C su banyosunda Yeri ve doku sindirim için bir gece bekletin. , Boncuk yıkamak için vorteks (10 sn) ve santrifüj (; 10 dakika süreyle 20 ° C&#39;de 2.000 rpm) Tween 80 (0.25%) ekleyin. , Süpernatantı 2-etoksietil asetat (1 mi) çözündürüldü, girdap ekleyin ve 1 saat boyunca bekletilir. Vortex süspansiyon ve santrifüj (2.000 rpm; (20 ° C&#39;de 10 dak) bir Hitachi F-2500 floresans spektrometresi kullanılarak bir küvet ve ölçülere floresans, yer içeren 2-etoksietil asetat sulu tabaka (eksitasyon 612/emission 618 çıkarın. ; Digilab, Holliston, MA). </li></ol>

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Çıkar çatışması ilan etti.

Test edilen bütün türlerde Sol pulmoner arter ligasyonu iskemik akciğer sağlam sistemik neovaskülarizasyon yol açar. Biz bir sıçan modelinde cerrahi yaklaşımın ayrıntıları sundu. Bizim sonuçlarımız vasküler döküm, histopatoloji tarafından üretilen, ve in vivo etiketleme bronşiyal arterler prolifere olduğunu göstermektedir ve akciğer parankimi serpmek. Böylece, bronşiyal anjiogenezin mekanizmaları kronik pulmoner tromboemboli insanlık durumunun paralellik bir hayvan modelinde okudu...

Akciğer Sistemik anjiyogenez iyi tanınır. Astım 2, interstisyel akciğer fibrozis 3, kanser 4, ve kronik pulmoner tromboemboli 5, sistemik damar ve akciğer parankimi akciğer proliferates çevreleyen ve işgal gibi hastalık durumlarında. Ancak, sistemik bu diferansiyel ziyade aktivasyonu pulmoner dolaşım incelemek için hayvan modelleri birkaçıdır. Belki de yetişkin memeli akciğerde sistemik neovaskülarizasyon en tekrarlanabilir modeli kronik pulmoner arter iskemi uyaran sonra oluştuğu olmasıdır. 5-7 insanlarda sol akciğer arter tıkanması olan cevap, köpekler 8, 9 domuz, koyun 10, kobay 11, sıçan 1, 12, 13, 14 ve fareler bronşiyal arter hem de interkostal arterlerin hızlı yayılmasıdır. Sistemik neovasculari sorumlu mekanizmalarpulmoner iskemi sonrası akciğer kıymetleştirme büyük ölçüde bilinmemektedir ve yaygın olarak çalışılmamıştır. Sol pulmoner arter tıkanıklığı sonrası sıçan bronş anjiogenezin zamanlı kurs dikkatli Weibel 1 histolojik çalışmada tarif edilmiştir. Bu iş rat uzanan, laboratuar bu süreçte önemli büyüme faktörleri yanı sıra, akciğer içinde bu neovaskülatür fizyolojik sonucu, her iki üzerine odaklanmıştır. Sonuçlar CXC kemokin CINC-3 erken iskemi sonrası ve CXCR2 bir nötralizan antikor ile sıçan tedavisinde yükselmiş göstermek, CINC-3 reseptör, anjiyogenez 13 azaltır. 14 gün pulmoner iskemi başlangıcından sonra yeni kurulan bronşiyal damar anlamlı artış protein geçirgenliği 15 ile anormal olduğu gösterilmiştir. Sol akciğer fonksiyonu difüzyon kapasitesi ve akciğer hacmi 15 bir azalma azalma normal bir gösteri değildi. Neovaskülatür co olsakronik akciğer iskemi sırasında akciğer dokusunun korunmasına ntributed, normal olmak ve solunum fonksiyon sürekli bir azalma katkıda bulunabilir olmadığı görülüyor. Belki de bu modelin en çok merak yönlerinden biri kan damarlarının genişlemesi mekansal dağılımı ile ilgilidir. Iskemiye bağlı akciğer parankimi içinde büyüme faktörlerinin serbest rağmen neovaskülatür nispeten büyük memba bronşiyal arterler kaynaklanır. Normal Bronşiyal arter aortadan küçük bir dal olarak ortaya çıkan ve karina de havayolu ağacı işgal etti. Böylece büyüme faktörleri arteriogenesis işleminin başlangıç ​​aşamasında indüklemek ile ilgili bir mekanizma açık değildir. Biz sıçan, insana benzer bir vasküler anatomisi, akciğer iskemi sırasında sistemik anjiyogenez sorumlu mekanizmaları incelemek için eşsiz bir fırsat sunuyor öneririz. Sol pulmoner arterde tam tıkanma insan deneklerde nadir bir olay olmasına rağmen,artmış bronşiyal vaskülarite benzer hastalarda indüklenebilirler görünüyor pulmoner arter tıkanıklığı 16 ne olursa olsun yeri ve boyutu. Böylece, sıçanlarda sol pulmoner arter ve anjiogenezin büyüklüğünü ölçmek için bir araç Arter için cerrahi yaklaşım ayrıntılı bir açıklamasını sağlar.

<table border="1"><tbody><tr><td> Reaktifler: </td><td> Şirket </td><td></td><td></td></tr><tr><td> buprenorfin hidroklorür, Puralube </td><td> Butler Schein </td><td></td><td></td></tr><tr><td> bupivicaine </td><td> APP İlaç </td><td></td><td></td></tr><tr><td> Povidon-İyot swabstick </td><td> Dynarex Şirketi </td><td></td><td></td></tr><tr><td> polipropilen sütür boyutu 6-0, 3/8 daire ters kesme iğne </td><td> Myco Tıp </td><td></td><td></td></tr><tr><td> PE20 boru </td><td> Becton Dickinson </td><td></td><td></td></tr><tr><td> 15 mikron kızıl polistiren florosfer </td><td> Invitrogen </td><td></td><td></td></tr><tr><td> 1 ml Hamilton cam şırınga </td><td> Hamilton Company </td><td> &lt;/ Td&gt; <td></td></tr><tr><td> Ekipman: </td><td></td><td></td><td></td></tr><tr><td> Genie Artı şırınga pompası </td><td> Kent Bilimsel </td><td></td><td></td></tr><tr><td> Floresans Spektrofotometre </td><td> Digilab </td><td></td><td></td></tr><tr><td> Kemirgen Vantilatör Model 683 </td><td> Harvard Apparatus </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> Tablo 1. Spesifik reaktifler ve ekipman Tablo. </td></tr></tbody></table>

angiogenesis in the ischemic rat lung

Erişkin akciğer sistemik bronşiyal arter ve pulmoner arterler üzerinden akan tüm venöz dönüşü hem perfüze edilir. Çoğu akciğer patolojilerinde, bu gelişmiş akciğer perfüzyon için bir ihtiyaca cevap veriyor ve sağlam neovaskülarizasyonu gösteren küçük sistemik damar olduğunu. Pulmoner arter tıkanıklığı ile uyarılan pulmoner vasküler iskemi insan hızlı bir sistemik arteryel anjiyogenez yanı sıra çok sayıda hayvan modellerinde neden olduğu gösterilmiştir. Sıçanlarda bronşiyal arter proliferasyon zaman dersin histolojik değerlendirmesi dikkatlice Weibel 1 tarafından tanımlanmış olmasına rağmen, yeni gemiler bu örgütlü büyümesi için sorumlu mekanizmalar net değildir. Biz bronşiyal neovaskülarizasyonu neden sıçan sol pulmoner arter iskemi oluşturulmadan cerrahi ayrıntıları. Anjiyogenez ölçüde kantitasyonu akciğer içinde iki vasküler yatak varlığı nedeniyle ek bir engel oluşturmaktadır. Yöntemleretiketli mikrosfer enjeksiyonları göre fonksiyonel anjiyogenez belirlemek için sağlanır.

Vasküler döküm: sıçan sol pulmoner arter iskemi etkilerinin sonuçları Şekil 1&#39;de gösterilmiştir. Gösterilen bronşiyal damar bir metakrilat döküm ve 28 gün LPAL sonra sol havayolu ağacının geniş vaskülarite olduğunu. Bu döküm elde etmek için, sistemik damar inen aorta retrograd bir metakrilat karışımı (kırmızı) enjekte edildi ve trakea kanüle ve beyaz bir silikon bazlı bir materyal enjekte edildi. Bu vasküler döküm akciğer bronş anjiyogenez olağanüstü bir gör...

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

Angiogenesis in the Ischemic Rat Lung

Akciğer sistemik bronşiyal arter ve pulmoner arterler hem perfüze edilir. Çoğu akciğer patolojilerinde, bu sağlam neovaskülarizasyonu gösterir küçük sistemik damar olduğunu. Pulmoner kan akımının kesilmesi tempolu bronşiyal anjiyogenez teşvik etmektedir. Biz bronşiyal neovaskülarizasyon teşvik sol pulmoner arter iskemi oluşturulmadan cerrahi ayrıntıları.

İskemik Sıçan Akciğer Angiogenezis

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

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Medicine

15.5K Görüntüleme.  Johns Hopkins University. Akciğer sistemik bronşiyal arter ve pulmoner arterler hem perfüze edilir. Çoğu akciğer patolojilerinde, bu sağlam neovaskülarizasyonu gösterir küçük sistemik damar olduğunu. Pulmoner kan akımının kesilmesi tempolu bronşiyal anjiyogenez teşvik etmektedir. Biz bronşiyal neovaskülarizasyon teşvik sol pulmoner arter iskemi oluşturulmadan cerrahi ayrıntıları.

Video: Angiogenesis in the Ischemic Rat Lung

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.

Manyetik Rezonans Görüntüleme, Volumetrik Bilgisayarlı Tomografi ve Ultrason Kullanımı Meme Kanseri Kemik metastazı bir Nude Rat Model Angiogenezis Multi-modal Görüntüleme

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

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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.

Sodyum Hyaluronat subretinal Enjeksiyon Kemirgenler Retina Dekolmanı Modeli

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.

Fare Akciğer Fibrin Gel implantasyonu Akciğer özel Anjiogenez Eğitim için

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.

İzole Yöntemi<em&gt; Ex vivo</emBir Rat Modelinde&gt; Akciğer Perfüzyon: Sıçan EVLP Programı Geliştirme Öğrenilmiş Dersler

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.

Fotokimyasal Bağlı Posterior İskemik Optik Nöropati Rat Modeli

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.

Sıçan Heterotopik Abdominal Kalp / Bir Ses yüklenen Yapılandırması Tek akciğer transplantasyonu

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.

Sıçan İskemik Cilt Yara Modeli Gösteri

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

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

Koroner Anjiyogenezin In Vitro Modeli

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...

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 ...

<meta charset="utf8"></head><body>Fare akciğer naklinde hücresel dinamiklerin gerçek zamanlı izlenmesi

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>Yazar Spotlight: İntravital Görüntüleme Yoluyla Akciğer Nakli İmmünolojisini İlerletmek

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.

Nakledilen Fare Akciğerinin İntravital İki Foton Mikroskobu

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