Bu eser Savunma Bakanlığı (DoD) bize ordu tıbbi araştırma ve gereç komutu (USAMRMC) Congressionally yönetmen tıbbi araştırma programı (CDMRP) (PR121604) ve ulusal kurumları artrit ve Musculoskeletal gelen hibe tarafından desteklenmiştir ve cilt hastalıkları (NIAMS), NIH R01 AR068332 Uma Sankar için.

Aşağıdaki yordam Indiana Üniversitesi okul tıp kurumsal hayvan bakımı ve kullanımı Komitesi (IACUC) onayı ile gerçekleştirildi. Tüm yaşam ameliyatları steril koşullarda NIH kurallar tarafından belirtildiği gibi yapıldı. Ağrı ve enfeksiyon riski uygun ilaçlar ve başarılı bir sonuç sağlamak için antibiyotik ile yönetilen.
1. anestezi ve hazırlık
<ol><li>Fare tartmak ve mayi (IP) rota bir yönetilen karışımı ketamin (100 mg/kg) ve xylazine (10 mg/kg) yolu ile ile anestezi. Boş bir kafes içinde fareyi getirin ve tamamen uyuşturulmuş kadar izlemek.</li>
	<li>Fareyi kullanarak bir ayak çimdik refleks sakin olun. Oftalmik merhem gözleri kurumasını engellemek için uygulanır.</li>
	<li>Kürk sağ bacak kaldırma. Bir iyot bazlı fırçalayın ve % 70 etanol ile cerrahi site aşağı silin. Diz merkezinde başlayan ve dairesel süpürme dışa doğru yapma cerrahi sitesi fırçalayın. Bu 3 x %70 etanol ile biten taze scrubs ile yineleyin.</li>
	<li>Buprenorfin hidroklorür analjezi (0.03 mg/kg) ameliyat öncesi bir doz subkutan için acil ameliyat sonrası ağrı yönetimi yönetmek.</li>
	<li>Steril Cerrahi pedi tarafından örtülü bir Isıtma yastık üzerinde fareyi getirin.</li>
</ol>2. cerrahi yaklaşım
Not: önce kırık, ağırlık ve damla yükseklik ampirik olarak belirli zorlanma, yaş ve cinsiyet ameliyat öncesi farelerin için tespit edilmelidir. Bu cerrahi işlem 10 haftalık C57BJ6 erkek fareler için optimize edilmiştir.
<ol><li>Sırtında fareyi getirin ve operatif bacak diz esnek. Bir neşter bıçak kullanarak, diz eklemi üzerinde ortalanmış bir 1,5 cm kesi yapmak.</li>
	<li>Yanal femur distal sonunda ortaya çıkarmak için forseps kullanarak diz kapağı yerinden. Retrograd bir şekilde ve Proksimal femur sonuna medüller Kanal uzunluğu aşağı trochlear oluk Merkezi 1.5 - inç uzun 25'lik paslanmaz çelik Hipodermik iğne yerleştirin. PIN doğru yerleşimini emin olmak için röntgen al. Not: İğne embolisinin için bir yol oluşturmak için fareyi sırt tarafında Office Communicator'dan çıkmalısınız.</li>
	<li>Distal uyluk, hub üzerinden girme ve eğimi fare dorsal tarafta çıkarken iğne mili ile 4 - inç uzun 36 kalibrelik tungsten embolisinin geçmek.</li>
	<li>Başarılı yerleşim embolisinin dikkatle 25'lik iğne vücudu ve embolisinin yerde tutarak hub üzerinde hafifçe çekerek çıkarın. X-ışını tarafından embolisinin yerleşimini onaylayın.</li>
	<li>391 g ağırlık etkisi disk (Şekil 1A) yukarıda 34.6 cm yükseklikten tutun. Öyle ki femur intertrochanteric ve supracondylar bölgelerinde destek örsler (Şekil 1B) dinlenmek ve ekstremite yan tarafındaki yükleme noktası (rakam 1 c karşı karşıya pozisyon yatay genelinde iki uyluk kemiği puan, destekleyen ). Ağırlığı bırakın ve dikkatle fare kırık hemen ardından aygıttan kaldırmak.</li>
	<li>Kırık konum X-ray tarafından onaylayın.</li>
	<li>24'lik paslanmaz çelik Hipodermik boru kırık uyluk dengelemeye embolisinin yerleştirin. Not: giriş noktası daha küçük çaplı bir iğne kullanılarak oluşturulan gibi bu uygulama bir güç gerektirebilir. Bu fark çapında etkili 24'lik çubuk Proksimal femur sonuna bir potansiyel göç engeller. Künt boru büyük trochanter kortikal kemik karşılar gibi el ile ekleme derinliği hissedilebilir.</li>
	<li>Çelik çubuk konumunu ve x-ışını tarafından kırık uyluk istikrar embolisinin kaldırmadan önce onaylayın.</li>
	<li>Tel kesiciler kullanarak femur distal uçta aşırı tüp kesti. Diz eklemi yerinden çıkar değil dikkatli olmak nazik bir aşağı kuvvet uygulamak için forseps kullanarak condyles yüzeyinin altında maruz kalan boru gömün.</li>
	<li>Forseps kullanarak diz kapağı yeniden konumlandırın. 5-0 absorbe dikiş kesi sitesiyle kapatın.</li>
</ol>3. ameliyat sonrası yönetim
<ol><li>Ameliyat, fareler, steril serum fizyolojik ile 500 µL ile onlara kendi ameliyat sonrası kurtarma yardım etmek için I.P. rota zerk.</li>
	<li>Cerrahiden uyandırmak kadar hayvanlar bir ısıtmalı kurtarma yatak üzerinde izlemek. Ayaktan sonra onları onların kafes için dönün.</li>
	<li>Yakından onlar düzgün şifa ve hareketlilik yeniden sağlamak için ameliyat sonrası birkaç gün fareler izlemeye devam edin. Buprenorfin hidroklorür analjezi (0.03 mg/kg) subkutan her 6 h 3 gün cerrahisi ve gerektikçe bundan sonra yönetmek. Onlar kemik ameliyat sonrası iyileşme zarar gösterildiği gibi non-steroid antienflamatuar ilaçlar (NSAİİ) kullanmaktan kaçının.</li>
</ol>

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Bu makale yazarları ifşa gerek yok. Bu el yazması bildirdi çalışmada kullanılan tüm malzeme için tam erişim üzerinde hiçbir kısıtlama yoktur yazarlar daha fazla durumu.

Bu cerrahi işlem standart kapalı femur kırığı farelerde oluşturmak için hedeftir. İnternal fiksasyon böylece bir bükülme İntramedüller çubuk kaçınarak kırık, nesil sonra yer alır bu modelin önemli bir avantaj sağlıyor. Belki de en önemli bir özelliği, bu iletişim kuralı bir standart transvers kırık femur midshaft, nesil gibidir kırık geometri uygulamalı bükme gücü ve arka bacak konumlandırma bağlıdır. Uygunsuz uyluk sırasında bükme an konumlandırma eğik veya parçalanmış kı...

Kırıklar için kas-iskelet sistemi meydana gelen en sık yaralanma bazıları ve 25 milyon $ her yıl Amerika Birleşik Devletleri1aşmak için öngörülen tedavi maliyeti de dahil olmak üzere büyük bir sosyo-ekonomik yük ile ilişkili olan, 2. Kırıklar çoğunluğu olaysız iyileşmek, şifa önemli kesintiler ve verimlilik kaybı ile ilişkili olsa da. Bütün kırıklar yaklaşık % 5-10 Neden gecikmiş şifa veya sendikasız, Yaş veya diğer altta yatan kronik hastalıklar, osteoporoz ve diyabet3,4,5gibi nedeniyle. Yok FDA onaylı farmakolojik tedaviler etkili kemik iyileşmesine katkıda bulunmak ve kurtarma süresini kısaltmak Şu anda mevcuttur.
Kırık iyileşmesi koordinasyon birden çok hücre tiplerinin içeren bir karmaşık ve son derece dinamik bir süreçtir. Bu nedenle, kemik rejenerasyon ile ilişkili hücresel ve moleküler olayların kapsamlı bir anlayış bu süreci hızlandırmak tedavi hedefleri tanımlaması için çok önemlidir. Olarak insan diğer hastalıklarla son derece uysal ve tekrarlanabilir hayvan modeli kurulması kemik iyileşme çalışmada çok önemlidir. Koyun ve domuz, gibi büyük hayvanlar kemik remodeling özellikleri ve biyomekanik uzatmaktı, benzer var ama pahalı, önemli ölçüde iyileşme zaman gerektirir ve genetik manipülasyon6için kolayca mükellef değildir. Öte yandan, sıçanlar ve fareler, gibi küçük hayvan modeller, taşıma, bakım, kısa üreme döngüsü ve daha kısa iyileşme süresi7düşük maliyetlerini hareket hızı da dahil olmak üzere birçok avantaj sunar. Ayrıca, fare genom tam olarak, hızlı düzenleme ve genetik varyantların nesil için izin sıralı. Böylece, fare insan hastalık, yaralanma, çalışma ve8onarmak için bir güçlü modeli sistemidir. İnsanlarda, osteoporoz ve diyabet gibi comorbidities gecikmiş bir iyileşme olasılığını artırmak. Varolan fare modelleri osteoporoz ve diyabet gibi comorbidities etkisi kemik yaralanma çalışma için kullanılabilir ve şifa vardır. Hastalarda osteoporoz9şifa bir kırık bir belirgin azalma kemik oluşumu daha sonraki aşamalarında var. Ovariektomili (OVX) fare hızlı kemik kaybı ve Gecikmeli kemik Postmenopozal Osteoporoz10,11' gözlemlenen benzer şifa sergi. Ayrıca, düşük kemik kitle fenotipleri ve insanlar11' de görülen bozulmuş kırık iyileşmesi birçok fare modelleri tip ı ve tip II diyabet taklit. Ayrıca, fare kırık modelleri Kallus içinde meydana gelen karmaşık biyolojik süreçlerin çalışmaya çok yönlü bir platform olarak hizmet ve kemik doku rejenerasyonu hızlandırmak roman tedavi stratejileri keşfetmek.
Kemik yapısı ve metabolizma, kemik kırığı kalır fareler ve insanlar, çok benzer şifa işlemindeki farklılıklara rağmen endochondral ve Intramembranous ossifikasyon kemik remodeling tarafından takip bir arada içeren. Endochondral ossifikasyon progenitör hücreler daha az mekanik olarak istikrarlı bölgelere nerede pituiter ve yumuşak bir Kallus üretmek için kıkırdak mineralize kondrosit ayırt etmek kırık boşluğu çevreleyen alımı içerir. İkinci dalga progenitör hücre Kallus sızmak ve yeni kemik matrisi12,13,14,15salgılar olgun dokusunu ayırt etmek. Intramembranous ossifikasyon sırasında periosteal ve endosteal yüzeylerinde ataları doğrudan dokusunu salgılayan Matrix'e ayırt etmek ve kırık boşluğu9,11,12 köprüleme kolaylaştırmak ,13. Birlikte, endochondral ve Intramembranous ossifications bir güçlü ikincil kemik mekanik yükler13,14 için elverişli oluşturmak için zaman içinde daha da yenilenmiş bir zor Kallus gelişimi sonucu ,15. Sağlıklı insanlarda iyileşme süreci yaklaşık 3 ay sadece 35 gün fareler16ile karşılaştırıldığında, alır.
Kırık iyileşmesi ya açık veya kapalı cerrahi modelleri17kullanarak yaygın olarak çalışılmıştır. Eleştirel boyutta nesil gibi açık cerrahi yaklaşımlar defekt veya osteotomi tamamlamak, sapmalar tarafından parçalanmış kırıklar neden azaltmak için geometri ve yaralanma konumunu standartlaştırmak. Şifa kez ertelendiği çünkü bir sendikaya arkasında yatan mekanizma çalışma için mükemmel bir model için kapalı kırık karşılaştırıldığında gibi ayak veya elde osteotomiler servis yapın. Ayrıca, bir katı eksternal fiksasyon osteotomized kemik rejenerasyon öncelikle Intramembranous ossifikasyon bağlıdır anlamı, stabilize etmek gereklidir. Açık cerrahi yaklaşımlar kilitleme çivi, PIN-müzik klipleri ve kilitleme tabak gibi aygıtlar kırık bacak için Aksiyel ve dönme istikrar sağlamak için kullanın; Ancak, bu tür cihazların pahalı ve önemli ölçüde ameliyatı18,19,20,21daha uzun süre gereklidir. Öte yandan, kapalı modelleri, endochondral şifa uyarmak yeterli istikrarsızlık için izin basit intramedüller fiksasyon cihaz bağları. Sonuç olarak, kapalı kırık modelleri kolayca bir sendikaya koşullarını taklit değil. Bunlar ucuz, kullanımı kolay ve ameliyat21,22,23yılında süresini en aza indirmek gibi İntramedüller iğne, çivi ve sıkıştırma vida, internal fiksasyon teknikleri avantajlı şunlardır. Bazı durumlarda önce kırığı İntramedüller pimleri eklenir ancak İntramedüller PIN eğilme bükülme veya bir değişken Kallus boyutu için katkıda bulunmak ve şifa kırık uyluk kemiği, deplasman yol açabilir. Bunlar neyin bir ağırlık diaphysis üzerinde bırakılan bir üç nokta bükme aygıtı kullanarak oluşturulur gibi geometri ve kırık konumunu kapalı modellerinde standart hale getirmek daha zordur. Ancak, uygun tekniği ile cerrahi bu yaklaşım hızlı ve tutarlı sonuçlar sunuyor. Ayrıca, kapalı kırık modeli yüksek kuvvet etkisi veya mekanik stres22neden kırıklar çalışmaya klinik bir araç olarak hizmet vermektedir.
Bu cerrahi protokolü üzerinden sıçanlar ve fareler22,24,25kırık uyluk dengelemek için bir İntramedüller PIN kullanarak önceden açıklanan yöntemleri uyarlanmıştır. İlk olarak, bir küçük çaplı bir İntramedüller iğne bir giriş noktası kurmak için intracondylar çentik eklenir ve bir embolisinin transvers kırık bir yerçekimi bağımlı üç nokta kullanarak femur midshaft, üreten önce tanıttı Aygıt bükme. Kapalı bir femur kırığı başarılı nesil, daha büyük çaplı bir İntramedüller çubuk kırık uyluk stabilize etmek için kılavuz tel üzerinden kurulmuştur. Bu yöntem çubuk sonrası kırık yerleşim için yaralı femur konumlandırılması ve en iyi duruma getirilmiş istikrar sağlayan gibi İntramedüller PIN bükülme sırasında kırık nedeniyle geciken şifa riski önler.

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the generation of closed femoral fractures in mice: a model to study bone healing

Kemik kırıkları hastaların yaşam kalitesini önemli ölçüde etkileyen ek olarak, büyük bir sosyo-ekonomik yük empoze. Verimli kemik iyileşmesine katkıda bulunmak tedavi stratejileri var olmayan ve yüksek talep vardır. Şifa kırıklar etkili ve tekrarlanabilir hayvan modelleri kemik rejenerasyon ile ilişkili karmaşık biyolojik süreçleri anlamak için ihtiyaç vardır. Kırık iyileşmesi pek çok hayvan modeller yıllar içinde oluşturulmuş olan; Ancak, fare kırık modelleri güçlü araçlar kemik iyileşmesi çalışmaya son zamanlarda ortaya çıkmıştır. Çeşitli açık ve kapalı modeller geliştirilmiştir, ancak fizyolojik ilgili bir şekilde hızlı ve tekrarlanabilir sonuçlar üretmek için basit bir yöntem olarak kapalı femur kırığı modeli öne çıkıyor. Bu cerrahi Protokolü tek taraflı kapalı femur kırığı farelerde oluşturmak ve femur kırığı sonrası istikrar bir İntramedüller çelik çubuk ekleyerek kolaylaştırmak için hedeftir. Daha fazla eksenel ve dönme istikrar aygıtların bir çivi veya bir vida teklif rağmen bir İntramedüller çubuk kullanımı kemik dokusunda yeni kusurları üreten veya yumuşak zarar olmadan tutarlı şifa sonuçları için yeterli bir istikrar sağlar doku. Radyografik görüntüleme Kallus oluşumu, kemikli Birliği ve sonraki kemik Kallus remodeling ilerlemesini izlemek için kullanılır. Kemik şifa sonuçları genellikle iyileşmiş kemik gücü ile ilişkili ve burulma test ile ölçülür. Yine de, kırık tamir ile ilişkili erken hücresel ve moleküler olayları anlama kemik doku rejenerasyonu çalışmada çok önemlidir. Kapalı femur kırığı modeli intramedüller fiksasyon ile farelerde kemik kırık iyileşmesi çalışma ve şifa hızlandırmak için tedavi stratejileri değerlendirmek için çekici bir platform olarak hizmet vermektedir.

Cerrahi işlem başarılı uygulanması ile radyografik görüntülemede takip. Anahtar adımlar bir İntramedüller iğne, bir kılavuz tel yerleşimini, femur midshaft ve uygun sabitleme bir İntramedüller çubuk (Şekil 2Aben - 2Aiv) ile enine bir kırık indüksiyon ekleme içerir. Kırık Kallus şifa ilerleme ile haftalık radyografik görüntüler kadar (2B rakam) ameliyatı takiben 28 g?...

Watch this Scientific Journal Video about The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing at JoVE.com

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

Fare kapalı femur kırığı kırık iyileşmesi eğitim için güçlü bir platform ve kemik rejenerasyon hızlandırmak için yeni tedavi stratejileri modelidir. Cerrahi bu iletişim kuralı tek taraflı kapalı femur kırık uyluk kemiği stabil İntramedüller çelik çubuk kullanarak farelerde oluşturmak için hedeftir.

Farelerde kapalı femur kırığı nesil: kemik şifa çalışması için bir Model

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies ...

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15.1K Görüntüleme.  Indiana University School of Medicine. Fare kapalı femur kırığı kırık iyileşmesi eğitim için güçlü bir platform ve kemik rejenerasyon hızlandırmak için yeni tedavi stratejileri modelidir. Cerrahi bu iletişim kuralı tek taraflı kapalı femur kırık uyluk kemiği stabil İntramedüller çelik çubuk kullanarak farelerde oluşturmak için hedeftir.

Video: The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

TSSB, Nöral Hematolojik ve Davranış Bağlantılı Revealing için Hayvan Modelinde Biyobelirteçler

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

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Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Yara Şifa murin Modeli

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Femoral Bone Marrow Aspiration in Live Mice

Serial sampling of the cellular composition of bone marrow (BM) is a routine procedure critical to clinical hematology. This protocol describes a detailed step-by-step technical procedure for an analogous procedure in live mice which allows for serial characterization of cells present in the BM. This procedure facilitates studies aimed to detect the presence of exogenously administered cells within the BM of mice as would be done in xenograft studies for instance. Moreover, this procedure allows for the retrieval and characterization of cells enriched in the BM such as hematopoietic stem and progenitor cells (HSPCs) without sacrifice of mice. Given that the cellular composition of peripheral blood is not necessarily reflective of proportions and types of stem and progenitor cells present in the marrow, procedures which provide access to this compartment without requiring termination of the mice are very helpful. The use of femoral bone marrow aspiration is illustrated here for cytological analysis of marrow cells, flow cytometric characterization of the hematopoietic stem/progenitor compartment, and culture of sorted HSPCs obtained by femoral BM aspiration compared with conventional marrow harvest.

This protocol describes a procedure for serial sampling of femoral bone marrow (BM) without requiring the sacrifice of mice. This procedure facilitates longitudinal studies of the BM composition of mice over time and provides serial access to cells within the BM for ex vivo and transplantation studies.

Canlı Farelerde Femoral Kemik İliği Aspirasyon

Serial sampling of the cellular composition of bone marrow (BM) is a routine procedure critical to clinical hematology. This protocol describes a detailed ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Farelerde Basit Kritik ölçekli Femoral Hata Modeli

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

Farelerde Yüz Sinir Aksotomi: Sakatlık Motonöron Yanıtı Eğitim için Bir Model

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Establishment of a Segmental Femoral Critical-size Defect Model in Mice Stabilized by Plate Osteosynthesis

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a model organism, the mouse has already been widely used in bone-related research. Large diaphyseal bone defect models in mice, however, are sparse and often use bone fixation which fills the bone marrow cavity and does not provide optimal mechanical stability. The objectives of the current study were to develop a critical-size, segmental, femoral defect in nude mice. A 3.5-mm mid-diaphyseal femoral ostectomy (approximately 25% of the femur length) was performed using a dedicated jig, and was stabilized with an anterior located locking plate and 4 locking screws. The bone defect was subsequently either left empty or filled with a bone substitute (syngenic bone graft or coralline scaffold). Bone healing was monitored noninvasively using radiography and in vivo micro-computed-tomography and was subsequently assessed by ex vivo micro-computed-tomography and undecalcified histology after animal sacrifice, 10 weeks postoperatively. The recovery of all mice was excellent, a full-weight-bearing was observed within one day following the surgical procedure. Furthermore, stable bone fixation and consistent fixation of the implanted materials were achieved in all animals tested throughout the study. When the bone defects were left empty, non-union was consistently obtained. In contrast, when the bone defects were filled with syngenic bone grafts, bone union was always observed. When the bone defects were filled with coralline scaffolds, newly-formed bone was observed in the interface between bone resection edges and the scaffold, as well as within a short distance within the scaffold. 
The present model describes a reproducible critical-size femoral defect stabilized by plate osteosynthesis with low morbidity in mice. The new load-bearing segmental bone defect model could be useful for studying the underlying mechanisms in bone regeneration pertinent to orthopaedic applications.

Although mouse models are invaluable tools for bone tissue engineering, models of long bone defects are sparse. This need motivated development of the present protocol which uses a locking plate with four screws and a dedicated jig to perform and stabilize a reproducible, femoral, critical-size defect with low morbidity.

Farelerde bir Segmental Femoral Kritik boyutlu Defekt Modelinde kurulması Plaka Osteosentez tarafından Stabilize

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a ...

An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming more commonly used in trauma research. They offer a large number of mutant strains and antibodies for the analysis of the molecular mechanisms behind the highly differentiated process of bone healing. To control the biomechanical environment, standardized and well-characterized osteosynthesis techniques are mandatory in mice. Here, we report on the design and use of an intramedullary nail to stabilize open femur osteotomies in mice. The nail, made of medical-grade stainless steel, provides high axial and rotational stiffness. The implant further allows the creation of defined, constant osteotomy gap sizes from 0.00 mm to 2.00 mm. Intramedullary locking nail stabilization of femur osteotomies with gap sizes of 0.00 mm and 0.25 mm result in adequate bone healing through endochondral and intramembranous ossification. Stabilization of femur osteotomies with a gap size of 2.00 mm results in atrophic non-union. Thus, the intramedullary locking nail can be used in healing and non-healing models. A further advantage of the use of the nail compared to other open bone healing models is the possibility to adequately fix bone substitutes and scaffolds in order to study the process of osseous integration. A disadvantage of the use of the intramedullary nail is the more invasive surgical procedure, inherent to all open procedures compared to closed models. A further disadvantage may be the induction of some damage to the intramedullary cavity, inherent to all intramedullary stabilization techniques compared to extramedullary stabilization procedures.

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

Femur Osteotomiler standardize Sabitleme için bir intramedüller Kilitleme Nail Fare Normal ve Arızalı Kemik Şifa Analiz

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

We describe a step-by-step method of performing direct pulp capping on mice teeth for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

Pulpa Değerlendirilmesi için Doğrudan Selüloz-kapatma Modelinin Geliştirilmesi Yara İyileşmesi ve Farelerde Onarım dentin oluşumu

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an increased use of mouse models in orthopedic research was noted, most probably because mouse models offer a large number of genetically-modified strains and special antibodies for the analysis of molecular mechanisms of fracture healing. To control the biomechanical conditions, well-characterized osteosynthesis techniques are mandatory, also in mice. Here, we report on the design and use of a closed bone healing model to stabilize femur fractures in mice. The intramedullary screw, made of medical-grade stainless steel, provides through fracture compression an axial and rotational stability compared to the mostly used simple intramedullary pins, which show a complete lack of axial and rotational stability. The stability achieved by the intramedullary screw allows the analysis of endochondral healing. A large amount of callus tissue, received after stabilization with the screw, offers ideal conditions to harvest tissue for biochemical and molecular analyses. A further advantage of the use of the screw is the fact that the screw can be inserted into the femur with a minimally invasive technique without inducing damage to the soft tissue. In conclusion, the screw is a unique implant that can ideally be used in closed fracture healing models offering standardized biomechanical conditions.

This protocol describes a minimally invasive osteosynthesis technique using an intramedullary screw for standardized stabilization of femur fractures, which can be used to analyze endochondral bone healing in mice.

Endochondral standart biyomekanik koşullar altında farelerde şifa kırık çözümlemek için minimal invaziv bir Model

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

Transvers aort daralma farelerde ikna etmek için kapalı-göğüs manken

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...