Yazarlar, teknik yardım ve sahne histoloji Christiane Pahrmann teşekkür ederim.

<html> 20g en az 8 ila 12 hafta arasında bir yaş ağırlığında Balb / C fareler Charles River (Sandhofer Weg 7, D-97.633 Sulzfeld) satın alınır. Fare, geleneksel koşulları, beslenen standart fare pelet ve su ad libitum altında muhafaza edilir . Indüksiyon odasına kullanarak (% 2) izofluran anestezisi fare ile. <ol><li> Boyun bölgesi ve göğüs kafesinin sol tarafında Tıraş ve% 80 etanol kullanılarak dezenfekte edin. </li><li> Sırtında fare koyun ve anestezi tutmak için burun ve ağız içinde bir facemask yerde. </li><li> Fare anesthesized yeterli olduğundan emin olmak için, kuyruğu ve arka ayaklarının kısma refleksleri kontrol edin. </li><li> Mikroskopik görünümü altında trakea kaplayan deri, kas ve doku ayıran orta hat servikal insizyon gerçekleştirin. </li><li> Trakea, glottis altında iki kartuş halkaları arasında doku içine endotrakeal tüp (Şekil 1) eklemek için küçük bir delik maruz kaldığında. Mikro cerrahi forecepts ile trakea kafatası parçası tutan endotrakeal tüp yerleştirin. Torasik hareketin her iki akciğerde iyi havalandırılmış olduğundan emin olmak için kontrol edin. </li><li> Solunum hızı (RR), 17 - 18cm H 2 O bir inspiratuar basınç ile, yaklaşık 110 dakikada olmalıdır Sağ tarafta uzanan, sol yan bakan, fareyi dikkatli bir şekilde açın. 3. ve 4. kaburga arasındaki leftsided torakotomi gerçekleştirin ve kanamayı önlemek için bir cauter kullanarak, dikkatli bir şekilde doku ve kas disseke. </li><li> Toraks dikkatle açın toraks açıldıktan sonra, herhangi bir keskin bir nesne ile akciğer dokunmadan, kalp bulmak. Şimdi perikardiyal kese kalp kapsayan bölümü kaldırmak. </li><li> LAD, pulmoner arter ve sol kulak kepçesi arasında yer alır. Tek sütür (Şekil 2) ile proksimal LAD Arter bir 8-0 Prolen sütür (Ethicon, Norderstedt, Almanya) kullanın. </li><li> 4. ve 5. kaburga arasındaki bir göğüs tüpü (28G, rüşvet kateter) yerleştirin. </li><li> Cilt kapatmak için kaburga ve 4-0 Prolen çalışan dikişler (Ethicon, Norderstedt, Almanya) uyum için 6-0 Prolen çalışan dikişler (Ethicon, Norderstedt, Almanya) kullanarak katmanlar halinde torasik insizyon kapatın. </li><li> Dikkatli bir şekilde (Şekil 3) 2 ml şırınga yardımı ile toraks boşaltın. Sırtında fare yerleştirin. Şimdi endotrakeal tüp ve 7-0 Prolen dikişler (Ethicon, Norderstedt, Almanya) kullanarak tek bir dikiş ile trakeal kartuş halkaları uyum. </li><li> Fare üzerinde yüz maskesi koyun ve dikişler (Ethicon, Norderstedt, Almanya) 4-0 Prolen kullanarak cilt yakın. </li></ol> Onaylama LAD-ligasyonu başarısını teyit etmek için farklı olasılıklar vardır. Hayvan euthanize gerek kalmadan sadece 150 lt kan troponin testi, cerrahi sonrası 6 ila 18 saat olabilir. Kan özelleştirilmiş bir troponin test kiti (TROP T Hassas, Roche, Mannheim) (Şekil 4) uygulanır. Troponin kas hücre aktin filamentler bir düzenleyici protein, kas üç tip farklı isotypes vardır. CTnI ve cTnT isotypes kalp bulunur ve doku hasarı olarak yayımlanır. Troponin T (cTnT) için standart bir test iki kalp spesifik monoklonal antikorlar, bir tuzakları kan örneği troponin dayalı, ikinci bir belirteçtir. Enfarkte alanda, 3 gün sonra (Şekil 5) makroskopik tespit edilebilir. Infarkt TTC (2,3,5-triphenyltetrazolium chloride) ile boyanarak önlemler dokusunun canlılığını değerlendirmek için kullanılır. Evans mavi boya (% 1,5, 1.0mL) fosfat tamponlu salin (PBS), iskemik miyokard alanını ölçmek için sol ventrikül boşluğuna enjekte edilir. Fare ötenazi ve kalp dilimler halinde hasat ve kesitli. Doku dilimleri 20dk, 37 ° C&#39;de% 1 TTC PBS solüsyonu, pH 7.4 inkübe edilir. Dokular 2-8 gecede PBS-tamponlu formalin% 10 sabit ° C TTC boya Evans mavi-negatif alanlarda (Şekil 6) ex vivo yönetilmektedir. Histoloji için fare ötenazi olmalı ve kalbin daha fazla işlem için gömülü olması gerekir. Kesitler yaptıktan sonra, slaytlar, H &amp; E (hematoksilen ve eozin) ile boyanmış veya fibrotik doku (Şekil 7) görselleştirmek için trikrom. <img src="/files/ftp_upload/1438/1438fig1.jpg" alt="Şekil 1" /> Şekil 1: Trakeal tüp. 1A: Açık trakea, ventral servikal insizyon sonrası. 1B: Trakea gırtlak kaudal iki kıkırdak halkaları açıldı. 1C: Trakeal tüp trakea içine sokulur. <img src="/files/ftp_upload/1438/1438fig2.jpg" alt="Şekil 2" /> Şekil 2: kalp sol taraflı torakotomi yoluyla maruz LAD bağlandı. t &quot;&gt; <img src="/files/ftp_upload/1438/1438fig3.jpg" alt="Şekil 3" /> Şekil 3: Toraks kapatıldıktan sonra, plevra drenaj yapılır. Fare anestezi altında devam etmektedir. <img src="/files/ftp_upload/1438/1438fig4.jpg" alt="Şekil 4" /> Şekil 4: troponin testi pozitif olduğu zaman iki satır bir kaç dakika sonra ortaya çıkar. <img src="/files/ftp_upload/1438/1438fig5.jpg" alt="Şekil 5" /> Şekil 5: eksplante kalp gösterir. 5A: iskemik alana sahip bir soluk beyaz renk ile kalbin makroskopik görünümü. 5B: enfarkte kalp Kesit, iskemik alanı sağ üst köşesinde gösterir.

<ol>
	<li>Atkins, B. Z., Hueman, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+cardiomyoplasty+improves+diastolic+properties+of+injured+Heart.">Cellular cardiomyoplasty improves diastolic properties of injured Heart.</a> J Surg Res. 85 (2), 234-242 (1999).</li><li>Jaquet, K., KT, K. r. a. u. s. e. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+myocardial+scar+size+after+implantation+of+mesenchymal+stem+cells+in+rats:+what+is+the+mechanism.">Reduction of myocardial scar size after implantation of mesenchymal stem cells in rats: what is the mechanism.</a> Stem Cells Dev. 14 (3), 299-309 (2005).</li><li>Joung, B., Kim, I. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+marrow+mononuclear+stem+cells+transplanted+in+rat+infarct+myocardium+improved+the+electrical+conduction+without+evidence+of+proarrhythmic+effects.">Bone marrow mononuclear stem cells transplanted in rat infarct myocardium improved the electrical conduction without evidence of proarrhythmic effects.</a> Yonsei Med J. 48 (5), 754-764 (2007).</li></ol>

Harçlar Bu çalışmada Üniversitesi Hastanesi Hamburg (MK, Kadın Habilitasyon Hibe), DFG (SS, SCHR992/3-1), ISHLT (TD, Araştırma Ödülü 2009) tarafından desteklenmiştir. Açıklama Bütün hayvanlar Laboratuar Hayvan Bakımı İlkeleri ile uyumlu insan bakım, hayvanlar üzerinde yapılan deneyler tarafından yayınlanan Laboratuvar Hayvanları Bakım ve Kullanım için Tıbbi araştırma ve Kılavuzu Ulusal Derneği tarafından belirlenen kurallara ve yönetmeliklere uygun olarak yapıldı Ulusal Sağlık Enstitüleri (Ulusal Sağlık yayın 85-23 Enstitüleri, 1985 revize). Bütün hayvanlar, Charles River Laboratuvarları (Sulzfeld Almanya) elde edilen ve hayvan bakım tesisleri Üniversitesi Hastanesi Hamburg Eppendorf tutuldu. Hayvanlar standart yem ve su ad libitum aldı.

Özetle, bu video LAD-ligasyonu modeli, adım-adım görselleştirir ve kolay uygulanabilir cerrahi işlemler olarak farelerde LAD-ligasyonu tanımlar.

lad-ligation: a murine model of myocardial infarction

Enfarktüsü ve miyokardiyal iskemi Araştırma modelleri, miyokard iskemisi, akut ve kronik pathobiological ve patofizyolojik süreçleri incelemek ve gelecekteki tedavi geliştirmek ve optimize etmek için gerekli. Miyokard iskemisi oluşturma İki farklı yöntemler laboratuar kemirgenler gerçekleştirilir. İlk yöntem oluşturmak için cryo enfarktüsü, hızlı ama yanlış bir tekniği, bir cryo-kalem (1-3) kalp yüzeyine uygulanır. Bu yöntemi kullanarak, bilim adamı cryo yara izi de geniş bir miyokard hasarı, miyokard enfarktüsü ile ilgili patofizyolojik yan etkileri gösterir oluşturulur, iskemi yol açar garanti edemeyiz. İkinci yöntem, sol ön inen arter (LAD) daimi ligasyonu. İşte LAD hemen görülebildiği bir iskemi oluşturan tek bir dikiş ile bağlandı. LAD kapatarak, daha fazla kan akımı, miyokard doku çevresindeki neredeyse etkilenmez iken bu alanda izin verilir. Bu cerrahi işlem, enfarktüs ilgili miyokard iskemisi meydana pathobiological ve patofizyolojik yönlerini taklit eder. Bu video tanıtılan yöntem LAD ligating bir fare enfarktüsü modeli cerrahi işlem göstermektedir. Bu model, kalp enfarktüsü pathobiological ve patofizyolojik yanı sıra Bağışıklık çalışmaları için uygundur. Gösterilen teknik, yüksek doğruluk sağlar ve histolojik bölümleri ile de ilişkilidir.

Watch this Scientific Journal Video about LAD-Ligation: A Murine Model of Myocardial Infarction at JoVE.com

LAD-Ligation: A Murine Model of Myocardial Infarction

Bu video, miyokard iskemisi pathobiological ve patofizyolojik süreçleri incelemek için hızlı ve güvenilir bir model nasıl kullanılacağını gösterir.

LAD-ligasyon murin Model: Miyokard Enfarktüsü

Research models of infarction and myocardial ischemia are essential to investigate the acute and chronic pathobiological and pathophysiological processes ...

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Research

JoVE Journal

Biology

44.3K Görüntüleme.  University Heart Center Hamburg. Bu video, miyokard iskemisi pathobiological ve patofizyolojik süreçleri incelemek için hızlı ve güvenilir bir model nasıl kullanılacağını gösterir.

Video: LAD-Ligation: A Murine Model of Myocardial Infarction

Murine Pancreatic Islet Isolation

Murin Pankreas Adacık İzolasyonu

Biyoloji

Murine Model of Hindlimb Ischemia 

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide. Medical therapies for symptomatic relief are limited. Surgical or endovascular interventions are useful for some individuals, but long-term results are often disappointing. As a result, there is a need for developing new therapies to treat PAD. The murine hindlimb ischemia preparation is a model of PAD, and is useful for testing new therapies. When compared to other models of tissue ischemia such as coronary or cerebral artery ligation, femoral artery ligation provides for a simpler model of ischemic tissue. Other advantages of this model are the ease of access to the femoral artery and low mortality rate. 
In this video, we demonstrate the methodology for the murine model of unilateral hindimb ischemia. The specific materials and procedures for creating and evaluating the model will be described, including the assessment of limb perfusion by laser Doppler imaging. This protocol can also be utilized for the transplantation and non-invasive tracking of cells, which is demonstrated by Huang et al.1.

Murine Model of Hindlimb Ischemia

The surgical procedure for induction of unilateral hindlimb ischemia is demonstrated, with confirmation of ischemia by laser Doppler perfusion imaging.

Murin Model Hindlimb İskemi

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide.  Medical therapies for ...

Murine Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with &gt; 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.
Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.

Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

Murin Böbrek Nakli Prosedürü

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

Rodent Working Heart Model for the Study of Myocardial Performance and Oxygen Consumption

Isolated working heart models have been used to understand the effects of loading conditions, heart rate and medications on myocardial performance in ways that cannot be accomplished in vivo. For example, inotropic medications commonly also affect preload and afterload, precluding load-independent assessments of their myocardial effects in vivo. Additionally, this model allows for sampling of coronary sinus effluent without contamination from systemic venous return, permitting assessment of myocardial oxygen consumption. Further, the advent of miniaturized pressure-volume catheters has allowed for the precise quantification of markers of both systolic and diastolic performance. We describe a model in which the left ventricle can be studied while performing both volume and pressure work under controlled conditions. 
In this technique, the heart and lungs of a Sprague-Dawley rat (weight 300-500 g) are removed en bloc under general anesthesia. The aorta is dissected free and cannulated for retrograde perfusion with oxygenated Krebs buffer. The pulmonary arteries and veins are ligated and the lungs removed from the preparation. The left atrium is then incised and cannulated using a separate venous cannula, attached to a preload block. Once this is determined to be leak-free, the left heart is loaded and retrograde perfusion stopped, creating the working heart model. The pulmonary artery is incised and cannulated for collection of coronary effluent and determination of myocardial oxygen consumption. A pressure-volume catheter is placed into the left ventricle either retrograde or through apical puncture. If desired, atrial pacing wires can be placed for more precise control of heart rate. This model allows for precise control of preload (using a left atrial pressure block), afterload (using an afterload block), heart rate (using pacing wires) and oxygen tension (using oxygen mixtures within the perfusate).

Isolated working heart models can be used to measure the effect of loading conditions, heart rate, and medications on myocardial performance and oxygen consumption. We describe methods for preparation of a rodent left heart working model that permits study of systolic and diastolic performance and oxygen consumption under various conditions.

Miyokard Performans ve Oksijen Tüketimi İnceleme Kemirgen Çalışma Kalp Modeli

Isolated working heart models have been used to understand the effects of loading conditions, heart rate and medications on myocardial performance in ways ...

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury

Chronic liver diseases, such as viral hepatitis, alcoholic liver disease, or non-alcoholic fatty liver disease, are characterized by continual inflammation, progressive destruction and regeneration of the hepatic parenchyma, liver progenitor cell proliferation, and fibrosis. The end-stage of every chronic liver disease is cirrhosis, a major risk factor for the development of hepatocellular carcinoma. To study processes regulating disease initiation, establishment, and progression, several animal models are used in laboratories. Here we describe a six-week time course of the choline-deficient and ethionine-supplemented (CDE) mouse model, which involves feeding six-week old male C57BL/6J mice with choline-deficient chow and 0.15% DL-ethionine-supplemented drinking water. Monitoring of animal health and a typical body weight loss curve are explained. The protocol demonstrates the gross examination of a CDE-treated liver and blood collection by cardiac puncture for subsequent serum analyses. Next, the liver perfusion technique and collection of different hepatic lobes for standard evaluations are shown, including liver histology assessments by hematoxylin and eosin or Sirius Red stainings, immunofluorescent detection of hepatic cell populations as well as transcriptome profiling of the liver microenvironment. This mouse model is suitable for studying inflammatory, fibrogenic, and liver progenitor cell dynamics induced through chronic liver disease and can be used to test potential therapeutic agents that may modulate these processes.

The Murine Choline-Deficient, Ethionine-Supplemented CDE Diet Model of Chronic Liver Injury

Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses.

Fare Kolin eksikliği, Ethionine desteklenmiştir (özgür) diyet kronik karaciğer yaralanma modeli

Chronic liver diseases, such as viral hepatitis, alcoholic liver disease, or non-alcoholic fatty liver disease, are characterized by continual ...

A Murine Model of Vertical Sleeve Gastrectomy

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. Bariatric surgery is currently the most effective long-term treatment for obesity. In addition to weight loss, bariatric surgery produces additional health benefits such as remission of type 2 diabetes, remission of hypertension, and decreased risk of developing certain types of cancer. The mechanisms beyond weight loss for these benefits remain incompletely defined. Therefore, animal models of bariatric surgery are being developed and validated to identify the mechanisms leading to these benefits, with the goal of improving understanding of gastrointestinal physiology and identifying new therapeutic targets. VSG has become the most commonly performed bariatric procedure in the clinic in the United States because it is highly effective at producing weight loss and metabolic improvement, and is simpler to perform than other bariatric procedures. Therefore, we have developed and validated a murine model of VSG. This murine VSG model recapitulates many of the effects of VSG seen in humans, including improved glucose and blood pressure regulation. The method is based on isolation of the stomach, ligation of gastric vessels, and removal of 70% of the stomach by transecting along the greater curvature of the stomach. We have successfully applied this surgical protocol to various genetically modified mouse lines to define the mechanistic contributors to the benefits of VSG. Furthermore, this murine VSG model has been combined with other surgical techniques, to achieve deeper mechanistic insight. Therefore, this is a simple and versatile model for studying gastrointestinal physiology and the health benefits of bariatric surgery.

The following describes the performance of vertical sleeve gastrectomy in mice. This is a type of weight-loss surgery that involves removal of approximately 70% of the stomach.

Dikey kol gastrektomi fare modeli

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer progression. How the chromosomal location and function of a centromere (i.e., centromere identity) are determined and participate in accurate chromosome segregation is a fundamental question. CENP-A is proposed to be the non-DNA indicator (epigenetic mark) of centromere identity, and CENP-A ubiquitylation is required for CENP-A deposition at the centromere, inherited through dimerization between cell division, and indispensable to cell viability.
Here we describe mass spectrometry analysis to identify ubiquitylation of EYFP-CENP-A K124R mutant suggesting that ubiquitylation at a different lysine is induced because of the EYFP tagging in the CENP-A K124R mutant protein. Lysine 306 (K306) ubiquitylation in EYFP-CENP-A K124R was successfully identified, which corresponds to lysine 56 (K56) in CENP-A through mass spectrometry analysis. A caveat is discussed in the use of GFP/EYFP or the tagging of high molecular weight protein as a tool to analyze the function of a protein. Current technical limit is also discussed for the detection of ubiquitylated bands, identification of site-specific ubiquitylation(s), and visualization of ubiquitylation in living cells or a specific single cell during the whole cell cycle.
The method of mass spectrometry analysis presented here can be applied to human CENP-A protein with different tags and other centromere-kinetochore proteins. These combinatory methods consisting of several assays/analyses could be recommended for researchers who are interested in identifying functional roles of ubiquitylation.

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

CENP-A ubiquitylation is an important requirement for CENP-A deposition at the centromere, inherited through dimerization between cell division, and indispensable to cell viability. Here we describe mass spectrometry analysis to identify ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A) protein.

EYFP etiketli CENP-A (EYFP-CENP-A) Ubiquitylation tanımlamak için Kütle Spektrometresi Analizi

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

Isolation of Human Ventricular Cardiomyocytes from Vibratome-Cut Myocardial Slices

The isolation of ventricular cardiac myocytes from animal and human hearts is a fundamental method in cardiac research. Animal cardiomyocytes are commonly isolated by coronary perfusion with digestive enzymes. However, isolating human cardiomyocytes is challenging because human myocardial specimens usually do not allow for coronary perfusion, and alternative isolation protocols result in poor yields of viable cells. In addition, human myocardial specimens are rare and only regularly available at institutions with on-site cardiac surgery. This hampers the translation of findings from animal to human cardiomyocytes. Described here is a reliable protocol that enables efficient isolation of ventricular myocytes from human and animal myocardium. To increase the surface-to-volume ratio while minimizing cell damage, myocardial tissue slices 300 &#181;m thick are generated from myocardial specimens with a vibratome. Tissue slices are then digested with protease and collagenase. Rat myocardium was used to establish the protocol and quantify yields of viable, calcium-tolerant myocytes by flow-cytometric cell counting. Comparison with the commonly used tissue-chunk method showed significantly higher yields of rod-shaped cardiomyocytes (41.5 &#177; 11.9 vs. 7.89 &#177; 3.6%, p &#60; 0.05). The protocol was translated to failing and non-failing human myocardium, where yields were similar as in rat myocardium and, again, markedly higher than with the tissue-chunk method (45.0 &#177; 15.0 vs. 6.87 &#177; 5.23 cells/mg, p &#60; 0.05). Notably, with the protocol presented it is possible to isolate reasonable numbers of viable human cardiomyocytes (9&#8211;200 cells/mg) from minimal amounts of tissue (&#60;50 mg). Thus, the method is applicable to healthy and failing myocardium from both human and animal hearts. Furthermore, it is possible to isolate excitable and contractile myocytes from human tissue specimens stored for up to 36 h in cold cardioplegic solution, rendering the method particularly useful for laboratories at institutions without on-site cardiac surgery.

Presented is a protocol for the isolation of human and animal ventricular cardiomyocytes from vibratome-cut myocardial slices. High yields of calcium-tolerant cells (up to 200 cells/mg) can be obtained from small amounts of tissue (&#60;50 mg). The protocol is applicable to myocardium exposed to cold ischemia for up to 36 h.

Vibratome-Cut Miyokardiyal Dilimlerden İnsan Ventriküler Kardiyomiyositlerin İzolasyon

The isolation of ventricular cardiac myocytes from animal and human hearts is a fundamental method in cardiac research. Animal cardiomyocytes are commonly ...

A Murine Model of a Burn Wound Reconstructed with an Allogeneic Skin Graft

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and require surgical debridement and skin grafting. Successful integration of the donor graft into a recipient wound bed depends on timely recruitment of immune cells, robust angiogenic response and new extracellular matrix formation. The development of novel therapeutic agents, which target some key processes involved in wound healing, are hindered by the lack of reliable preclinical models with optimized objective assessment of wound closure. Here, we describe an inexpensive and reproducible model of experimental full thickness burn wound reconstructed with an allogeneic skin graft. The wound is induced on the dorsum surface of anaesthetized inbred wild type mice from the BALB/C and SKH1-Hrhr backgrounds. The burn is produced using a brass template measuring 10 mm in diameter, which is preheated to 80 &#176;C and delivered at a constant pressure for 20 s. Burn eschar is excised 24 hours after the injury and replaced with a full thickness graft harvested from the tail of a genetically similar donor mouse. No specialized equipment is required for the procedure and surgical techniques are straightforward to follow. The method may be effortlessly implemented and reproduced in most research settings. Certain limitations are associated with the model. Due to technical difficulties, the harvest of thinner split thickness skin grafts is not possible. The surgical method we describe here allows for the reconstruction of burn wounds using full thickness skin grafts. It may be used to carry out preclinical therapeutic testing.

The aim of this study was to develop a murine model of burn wound healing. A thermal burn was induced on the dorsal skin of mice using a preheated brass template. Burned tissue was debrided and overlaid with a skin graft harvested from the tail of a genetically similar donor mouse.

Allojenik Deri Grefti ile Yeniden Yapılandırılan Yanık Yarasının Murine Modeli

Trivial superficial wounds heal without complications by primary intention. Deep wounds, such as full thickness burns, heal by secondary intention and ...

The Establishment of a Murine Mandibular Molar Extraction Socket Healing Model

This study introduces the development of a molar extraction model in the murine mandible to provide a practicable model for studying alveolar bone regeneration and intramembranous ossification. C57/J6 mice were used to extract the mandibular first molar to establish this model. They were euthanized, and the bilateral mandibles harvested, at 1 week and 4 weeks post-surgery, respectively. Subsequent serial stereoscopic harvest, histological assessment, and immunofluorescence staining were performed to demonstrate successful surgery. Immediately after surgery, the stereoscopic images displayed an empty extraction socket. The hematoxylin and eosin (H&#38;E) at 1 week and Masson staining at 4 weeks post-surgery showed that the area of the original root was partially and fully filled with bone trabeculae, respectively. The immunofluorescence staining showed that, compared with the homeostasis side, the Sp7 expression increased at 1 week post-surgery, suggesting vigorous osteogenesis in the alveolar fossa. All these results demonstrated a practicable murine tooth extraction socket healing model. Upcoming studies revealing the mechanisms of jawbone defect healing or socket healing could adopt this method.

This protocol demonstrates step-by-step details of how to extract the mandibular first molar in the mouse. It provides an alternative method for researchers focusing on jawbone healing and regeneration.