Name: the generation of closed femoral fractures in mice: a model to study bone healing
Uploaded: 2018-08-16T14:30:00.000+00:00
Duration: 5 min 53 s
Description: Watch this Scientific Journal Video about The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing at JoVE.com

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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Bone Tissue and the Skeletal System

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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

Pseudofracture: An Acute Peripheral Tissue Trauma Model

Following trauma there is an early hyper-reactive inflammatory response that can lead to multiple organ dysfunction and high mortality in trauma patients; this response is often accompanied by a delayed immunosuppression that adds the clinical complications of infection and can also increase mortality.1-9 Many studies have begun to assess these changes in the reactivity of the immune system following trauma.10-15 
Immunologic studies are greatly supported through the wide variety of transgenic and knockout mice available for in vivo modeling; these strains aid in detailed investigations to assess the molecular pathways involved in the immunologic responses.16-21
The challenge in experimental murine trauma modeling is long term investigation, as fracture fixation techniques in mice, can be complex and not easily reproducible.22-30
This pseudofracture model, an easily reproduced trauma model, overcomes these difficulties by immunologically mimicking an extremity fracture environment, while allowing freedom of movement in the animals and long term survival without the continual, prolonged use of anaesthesia. The intent is to recreate the features of long bone fracture; injured muscle and soft tissue are exposed to damaged bone and bone marrow without breaking the native bone.
The pseudofracture model consists of two parts: a bilateral muscle crush injury to the hindlimbs, followed by injection of a bone solution into these injured muscles. The bone solution is prepared by harvesting the long bones from both hindlimbs of an age- and weight-matched syngeneic donor. These bones are then crushed and resuspended in phosphate buffered saline to create the bone solution.
Bilateral femur fracture is a commonly used and well-established model of extremity trauma, and was the comparative model during the development of the pseudofracture model. Among the variety of available fracture models, we chose to use a closed method of fracture with soft tissue injury as our comparison to the pseudofracture, as we wanted a sterile yet proportionally severe peripheral tissue trauma model. 31
Hemorrhagic shock is a common finding in the setting of severe trauma, and the global hypoperfusion adds a very relevant element to a trauma model. 32-36 The pseudofracture model can be easily combined with a hemorrhagic shock model for a multiple trauma model of high severity. 37

Pseudofracture, a reproducible murine model of sterile musculoskeletal trauma, allows for evaluation of late term post-traumatic immune responses. This article describes the procedural execution of the model step by step, including the potential for experimental model combinations to permit study of multiple trauma.

Pseudofracture: Akut Periferik Doku Travma Modeli

Following trauma there is an early hyper-reactive inflammatory response that can lead to multiple organ dysfunction and high mortality in trauma patients; ...

Tıp

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 ...

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 ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

Vivo Fare uyluk için bir MRI uyumlu Osteosentez aygıtı kullanma farelerde kemik iyileşme sırasında kırık Kallus gelişimi değerlendirme

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes. Here, we introduce a clinically relevant mouse model of tibial fracture. The postoperative changes in these mice mimic some of the cognitive impairments commonly observed after routine orthopedic surgery in humans. Briefly, an incision is performed in the right hind limb under strictly aseptic conditions. Muscles are disassociated, and a 0.38-mm stainless steel pin is inserted into the upper crest of the tibia, inside the intramedullary canal. Osteotomy is then performed, and the wound is stapled. We have used this model to investigate the effects of surgical trauma on postoperative neuroinflammation and behavioral changes. By applying this fracture model in combination with parabiosis, a surgical model in which 2 mice are anastomosed, we have studied cells and secreted factors that systemically rejuvenate organ function and tissue regeneration after injury. By following our step-by-step protocol, these models can be reproduced with high fidelity, and can be adapted to interrogate many biologic pathways that are altered by surgical trauma.

This protocol describes a mouse model of orthopedic surgery that has been used to study mechanisms of postoperative neuroinflammation and behavioral changes, and when combined with parabiosis, to study tissue regeneration during aging.

Ameliyat sonrası bilişsel işlev bozukluğu ve doku rejenerasyonu çalışmaya ortopedik cerrahi bir fare modeli

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

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 ...

Protocol for Developing a Femur Osteotomy Model in Wistar Albino Rats

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. Osteoanabolic drugs have the potential to augment the repair of fractures but have constraints like high costs or undesirable side effects. The bone healing potential of a drug can initially be determined by in vitro studies, but in vivo studies are needed for the final proof of concept. Our objective was to develop a femur osteotomy rodent model that could help researchers understand the development of callus formation following fracture of the shaft of the femur and that could help establish whether a potential drug has bone healing properties. Adult male Wistar albino rats were used after Institutional Animal Ethics Committee clearance. The rodents were anesthetized, and under aseptic conditions, complete transverse fractures at the middle one-third of the shafts of the femurs were created using open osteotomy. The fractures were reduced and internally fixed using intramedullary K-wires, and secondary fracture healing was allowed to take place. After surgery, intraperitoneal analgesics and antibiotics were given for 5 days. Sequential weekly x-rays assessed callus formation. The rats were sacrificed based on radiologically pre-determined time points, and the development of the fracture callus was analyzed radiologically and using immunohistochemistry.

Here, we present a protocol to iatrogenically fracture the shaft of the femur of Wistar albino rats and follow up on the development of the callus. This femur osteotomy model can help researchers evaluate the process of fracture healing and to study how a drug could influence fracture healing.

Wistar Albino Sıçanlarında Femur Osteotomi Modeli Geliştirme Protokolü

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. ...

Transverse Fracture of the Mouse Femur with Stabilizing Pin

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test whether a gene, gene product or drug influences bone repair because murine bones recapitulate the stages observed during human fracture healing. When a mouse or human breaks a bone, an inflammatory response is initiated, and the periosteum, a stem cell niche surrounding the bone itself, is activated and expands. Cells residing in the periosteum then differentiate to form a vascularized soft callus. The transition from the soft callus to a hard callus occurs as the recruited skeletal progenitor cells differentiate into mineralizing cells, and the bridging of the fractured ends results in the bone union. The mineralized callus then undergoes remodeling to restore the original shape and structure of the healed bone. Fracture healing has been studied in mice using various injury models. Still, the best way to recapitulate this entire biological process is to break through the cross-section of a long bone that encompasses both cortices. This protocol describes how a stabilized, transverse femur fracture can be safely performed to assess healing in adult mice. A surgical protocol including detailed harvesting and imaging techniques to characterize the different stages of fracture healing is also provided.

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

Fare Femurunun Stabilize Pin ile Enine Kırılması

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test ...

Biyoloji

Establishing a Diaphyseal Femur Fracture Model in Mice

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age and overall health status of the patient, failures can occur, leading to delayed union or nonunion. Due to the increasing number of fractures resulting from high-energy trauma and aging, the development of innovative therapeutic strategies to improve bone repair based on the combination of skeletal/mesenchymal stem/stromal cells and biomimetic biomaterials is urgently needed. To this end, the use of reliable animal models is fundamental to better understanding the key cellular and molecular mechanisms that determine the healing outcomes. Of all the models, the mouse is the preferred research model because it offers a wide variety of transgenic strains and reagents for experimental analysis. However, the establishment of fractures in mice may be technically challenging due to their small size. Therefore, this article aims to demonstrate the procedures for the surgical establishment of a diaphyseal femur fracture in mice, which is stabilized with an intramedullary wire and resembles the most common bone repair process, through cartilaginous callus formation.

This protocol describes a surgical procedure for the establishment of a diaphyseal fracture in the femur of mice, which is stabilized with an intramedullary wire, for fracture healing studies.

Farelerde Diafiz Femur Kırığı Modelinin Kurulması

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age ...