Name: an intramedullary locking nail for standardized fixation of femur osteotomies to analyze normal and defective bone healing in mice
Uploaded: 2016-11-13T15:00:00.000+00:00
Duration: 7 min 41 s
Description: Watch this Scientific Journal Video about An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice at JoVE.com

Bu çalışma RISystem AG, İsviçre&#39;nin Davos tarafından desteklenmiştir.

Tüm işlemler IACUC onaylı ve kurumsal ilkelerine uygun (Landesamt für Verbraucherschutz, Zentralstelle Amtstierärztlicher Dienst, Saarbrücken, Almanya) izledi. Ağrı ve enfeksiyon önleme deneyleri yapılacak ülkenin ve kurumun ilgili kurallar ile uyum içinde olmalıdır. İmplantlar ve Cerrahi Aletler hazırlanması 1. <ol><li> mikrocerrahi alet kutusundan bir neşter bıçak (boyut 15), küçük hazırlık makas, ince forseps, soyunma forseps, küçük kerpeten, 24 ayar (G) ve 27 G iğne olmayan bir rezorbe 5-0 sütür, ve bir iğne tutucu seçin . </li><li> Intramedüller çivi, kilitli iğne, özel amaçlı cihaz, Gigli gördüm, Gigli için şablon gördüm, merkezleme matkap ucu (1 mm çapında), matkap ucu (0.3 mm çapında), ve el matkabı ambalajından (Şekil 2;) Malzeme listesine bakınız. NOT: intramedullary tırnak (0.8 mm çap, 15.7 mm uzunluk) femur içine retrograd implantasyon tıbbi kalite paslanmaz çelikten yapılmış bir intramedüller kilitli çivi olduğunu. Tırnak bir yakın ipliği (4 mm uzunluk) ve eksenel ve dönel stabilite elde etmek için birbirine pimlerin sokulması (0.3 mm çap) için iki delik (Şekil 1) yer alır. </li><li> (130 ° C, 25 dk buhar sterilizasyonu) 5 dakika boyunca bir dezenfekte edici solüsyon (örneğin,% 96 alkol) için implantlar ve tüm cerrahi aletler Açığa veya sterilize. Dezenfeksiyon veya Sterilizasyon sonrasında, steril bir operasyon bez üzerine aletleri yerleştirin. Küçük hayvan operasyon masasına doğrudan bitişik steril çalışma bez yerleştirin. </li></ol> 2. Hayvanlar, Anestezi ve Analjezi <ol><li> çalışma ve ele alınması soru için gerekli farelerin suşu, yaş ve cinsiyet seçin. Not: 14-haftalık erkek CD-1 fareler, bu çalışmada 12 için kullanıldı. tırnak impla için35 gr - ntation, hayvanların ideal vücut ağırlığı 25&#39;tir. </li><li> 15 mg / kg ksilazin 75 mg / kg ketamin intraperitoneal enjeksiyonu ile farelere anestezi. ayak tutam tarafından anesthetization onaylayın. anestezi sırasında kurumasını hayvanların gözleri korumak için göz yağ sürün. Anestezi indüksiyonu sonrasında, vücut sıcaklığını sabit tutmak için bir ısı lambası altında fare yerleştirin. </li><li> Ameliyat sonrası 3 kadar ameliyat öncesi gün 1 analjezi için içme suyu (2.5 mg / 100 mi) içindeki tramadol hidroklorür uygulanır. </li></ol> 3. Cerrahi Prosedür ve Tırnak İmplantasyon <ol><li> Ameliyattan önce, tüm sağ arka bacak tıraş ve tüy dökücü krem ​​uygulayın. 5 dakika sonra, krem ​​ve suyla bacak temizleyin. (130 ° C, 25 dk buhar sterilizasyonu) bir dezenfekte edici solüsyon (örneğin,% 96 alkol) için implantlar ve tüm cerrahi aletler Açığa veya sterilize. </li><li> aseptik koşullar altında, ön anlaşması yerKüçük hayvan operasyon masasında sırtüstü pozisyonda se. Diz kondillerin bir anterior yaklaşım sağlamak için sağ diz bükün. neşter bıçak kullanarak sağ diz 5 mm medial parapatellar insizyon gerçekleştirin. </li><li> ince forseps ile patellar ligament kaldırın ve neşter bıçağı ile dikkatlice ligaman seferber. Ardından, femur interkondiler çentik ortaya çıkarmak için neşter bıçağı ile yanal patellayı kaydırır. </li><li> kemik iliği boşluğuna ulaşana kadar delerek interkondiler çentik açın. <ol><li> matkap merkezleme 1 mm ile femur eksenine offset 45º ile sondaj başlayın. Yavaş yavaş o femur kemik eksenini paralel kadar sondaj esnasında matkabın yönünü değiştirmek. kemik iliği boşluğuna ulaşılırsa sondaj durdurun. </li></ol></li><li> interkondiler çentik at kemiği açtıktan sonra, femur tüm uzunluğu boyunca intramedüller boşluğuna 24 G iğne takın. INTRAME delmekelle 24 G iğne döner hareketleri ile femur dullary boşluğu. 24 G iğne çıkarın ve intramedüller boşluğuna ince 27 G iğne takın. proksimale büyük trokanter femur kortikal kemiğe perfore ileri iğne itin. </li><li> Femur 27 G iğne çıkarın. çivinin distal uç kondillerin seviyesine ulaşıncaya kadar el matkabı kullanarak, sürekli dönme ve eksenel basınç altında interkondiler çentik ile intramedüller çivi implant. Not: tırnağın uzak ucu küçük bir işareti tespit edilebilir. </li><li> sol lateral pozisyonda fare yerleştirin. cerrahi femur orta cisim ortaya koymak için kalça eklemine diz ekleminin lateral femur cisim kısmı boyunca neşter bıçak kullanarak bir uzunlamasına deri kesisi gerçekleştirin. </li><li> Küçük hazırlanması makas kullanarak, fasya bölünmüş ve yan taraftan femur ekseni yönünde kasları yayıldı.femur cisim parçası açığa çıkana kadar kasları yayıldı. siyatik siniri korumak. <ol><li> soyunma forseps ile kemik zayıflatarak femur tüm çevresi hazırlayın. Ardından, giyinme forseps yayarak kasları geri çekin ve femur maruz kalmaktadır. </li></ol></li><li> çivinin uzak ucuna hedefleme cihazı monte edin. o tırnağın adaptör flanşı verdiği kadar cihazı ilerlemek ve femur anterolateral konumda amaçlayan cihazı açın. </li><li> Bir proksimal ve distal kilitleme pimi ile çivi kilitlenebilir. <ol><li> Proksimal kilitleme pimi ile başlayın. </li><li> El matkap içine ortalama matkap ucu (1 mm çapında) yerleştirin. yakın kilitleme deliği pozisyonunda kemik havşa. Not Havşa olarak, küçük bir kavite kemiği ile delmeden bakan kortikal kemik oluşturulur. Bu boşluğu, geliştirilmiş merkezleme ve ince matkap kılavuz (0.3 mm çap) sağlardaha sonra kullanılan. </li><li> El matkap içine matkap ucu (0.3 mm çap) yerleştirin. amaçlayan cihazı kullanılarak, kaplama ve kaçırdı kortikal kemiğin (bikortikal) hem aracılığıyla delik açın. amaçlayan cihaz üzerinden ilk kilitleme pimini takın. en kısa sürede birbirine tork elde edilir kapalı olarak birbirine pim tahrik mili makası. </li><li> Distal kilitleme pimi için bu yordamı yineleyin. </li></ol></li><li> diafiz osteotomiyi gerçekleştirin. <ol><li> İki birbirine pimleri arasındaki yan tarafında amaçlayan cihaza testere kılavuzunu takın. Gigli tuzlu su ile sürekli sulanan gördüm ile Ardından, kemik gördüm. osteotomi tamamlandıktan sonra, kemiğe yakın bir ucunda kesim hattını. yumuşak dokuya zarar vermemek için dikkatli testereyi çıkarın. </li></ol></li><li> işaretli hattında intramedüller çivi kalan milini off klibi, küçük kerpeten ile, nişan cihazını çıkarın ve. </li><li> lat kas katmanları kapatıneral femur site ve tek dikişlerle deri kapatılması gerçekleştirin. diz ön yerinde, Patellayı yeniden konumlandırmak ve tek sütür ile kaslara patella tendonu sabitleyin. yanı sıra bu yarayı kapatmak için tek dikişlerle kullanın. </li><li> Anestezi kurtarmak kadar ısı lambası altında hayvanların tutun. Onlar ventral yatma muhafaza edilmesi için yeterli bilinci yerine kadar sahipsiz hayvanları bırakmayın. hayvan barınağında tek kafeslere hayvanların dönün. </li><li> Her gün özenle hayvanları izleyin. İlk üç gün boyunca postoperatif analjezi koruyun. Ameliyat sonrası 4. günde, hayvanlar hala acı kanıt gösterebilir, eğer seslendirme, huzursuzluk, hareketlilik olmaması, damat başarısızlık, anormal duruş ve çevresi normal ilgi eksikliği ile gösterildiği gibi, analjezi devam edin. hayvanlar ağrı serbest zaman analjezi sonlandırın. </li></ol>

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Romano Matthys is an employee of RISystem AG, Davos, Switzerland, which produces the implants and implant-specific instruments. The other authors have no conflicts of interest.

Cerrahi tekniğin en kritik adımlar çivi, nişan alma aleti ve pimleri doğru konumlandırma vardır. Tırnak kondilin seviyesinde diz eklemine çivinin bir çıkıntı diz (Şekil 3 A) hareketini kısıtlayabilir için, tırnağın uzak ucunda belirgin girinti tamamen yerleştirilmiş olması gerekir. Bu nedenle, femur boyutu ve buna bağlı olarak, hayvan vücut ağırlığına, dikkate alınmalıdır. Cerrah ayrıca tırnağın adaptör flanş bağlama amaçlayan cihazın nihai pozisyonuna özel di...

Kemik iyileşmesi biyolojisi, hücre ve küremsi kültürleri kullanılarak, in vitro incelenmiştir olabilir, ama aynı zamanda, hayvan çalışmaları kullanılarak yaklaşımları in vivo gerektirir. Büyük hayvan deneyleri hala klinik öncesi test önemli bir rol oynasa da, ürün veya hipotezlerin erken evre testi son 10 yıl içinde değişti ve günümüzde genellikle küçük hayvan modellerinde 1 yapılır. Bu anahtar, çeşitli nedenlerle uygulandı. Üretimi ve fareler ve sıçanlar bakımı daha ucuz domuz, koyun ile karşılaştırılmaktadır. Buna ek olarak, küçük hayvanlar kronik deneyler büyük serilerin performansını kolaylaştırmak her ikisi de kısa üreme süreleri ve daha kısa normal iyileşme dönemleri var. Son olarak, gen hedefli hayvan ve özel antikorların mevcudiyeti kemik iyileşmesi moleküler mekanizmaların analizi sağlar. Daha önce büyük hayvan modellerinde osteosentez tekniklerinin kullanıldığı Bununla birlikte, asgari variat ile tercüme edilebilir<html> insan ya da veterinerlik klinik hasta bakım, küçük boyutlu sıçan ve farelerde gelişimi ve osteosentez tekniklerinin uygulanmasında kullanılan benzer prosedürlere iyon zor olduğu ortaya çıktı. İyi biyomekanik ortam anlamlı süreci 2 şifa kemik etkiler bilinmektedir. İnsanlarda kırık iyileşmesi bilindiği gibi, küçük hareketlerini daha az rijit fiksasyon sonra rijit fiksasyon ve endokondral kemikleşme sonra intramembranous kemikleşme de dahil olmak üzere şifa farklı modları, kırık stabilizasyonu sonucu farklılıklar. Tam eksenel veya dönel instabilite iyileşme süreci geciktirebilir veya non-birlikleri 3 neden olabilir. Dolayısıyla, farelerde ve sıçanlarda sofistike implant sistemleri ve osteosentez teknikleri geliştirmek için gerekli olduğunu hissediyorum. Bu şekilde, biyomekanik koşullar iyileşme sürecini analiz ederken geçerli sonuçlar garanti uygun standardize edilebilir. e_content &quot;&gt; son derece gelişmiş murin sabitleyici önemli bir bölümü, son birkaç yıl içinde ortaya olmasına karşın, en yaygın olarak kullanılan bir teknik yine de basit intramedüller pimdir. Bu tekniğin başlıca dezavantajı, dönme ve eksenel olmaması kararlılık 4.. rotasyonel ve eksenel istikrarı geliştirmek için, bir intramedüller vida farelerde 5 femur kırıkları stabilize etmek için kullanılmaya başlandı. Ancak, vida ile tespit dolayı kemik parçaları arasında temas ve sıkıştırma ihtiyacını kemik kusurlu iyileşmesini analiz etmek için kullanılamaz rotasyonel istikrarı korumak için sipariş. Çivi kilitleme intramedüller yüksek eksenel ve basit pin ve intramedüller vida 4 ile karşılaştırıldığında rotasyonel stabiliteyi sunuyor. Çünkü Gigli için kılavuzun mümkün bir yüksek tekrarlanabilir femur osteotomisi, gördüm ve tanımlanmış boşluk boyutları oluşturma yeteneği, hem normal hem de bon analizi için izin verire iyileşmesi ve kemik arızalı iyileşmesi 6. Nedeniyle birbirine pimlerin sokulması için, çivi kilitleme intramedüller tüm ağırlığını taşıyan bile, tüm iyileşme sürecinde sürekli bir boşluk boyutu garanti eder. Burada, tasarım ve intramedüller kilitleme çivi uygulaması hakkında rapor, hem de avantajları ve normal ve gecikmiş kemik iyileşmesi üzerine deneysel çalışmalarda dezavantajları olduğu gibi.

<table><tbody><tr><td>MouseNail</td><td>RISystem AG</td><td>221,122</td><td></td></tr><tr><td>MouseNail aiming device</td><td>RISystem AG</td><td>221,201</td><td></td></tr><tr><td>MouseNail interlocking pin</td><td>RISystem AG</td><td>221,121</td><td></td></tr><tr><td>Centering bit</td><td>RISystem AG</td><td>592,205</td><td></td></tr><tr><td>Drill bit</td><td>RISystem AG</td><td>590,200</td><td></td></tr><tr><td>Gigli wire saw</td><td>RISystem AG</td><td>590,100</td><td></td></tr><tr><td>Suture (5-0 Prolene)</td><td>Ethicon</td><td>8614H</td><td></td></tr><tr><td>Forceps</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BD520R</td><td></td></tr><tr><td>Dressing forceps</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BJ009R</td><td></td></tr><tr><td>Scissors</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BC100R</td><td></td></tr><tr><td>Needle holder</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BM024R</td><td></td></tr><tr><td>24 G needle</td><td>BD Mircolance 3</td><td>304100</td><td></td></tr><tr><td>27 G needle</td><td>Braun Melsungen AG</td><td>9186182</td><td></td></tr><tr><td>Scalpel blade size 15</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>16600525</td><td></td></tr><tr><td>Pincers</td><td>Knipex</td><td>7932125</td><td></td></tr><tr><td>Heat radiator</td><td>Sanitas</td><td>605.25</td><td></td></tr><tr><td>Depilatory cream</td><td>Asid bonz GmbH</td><td>NDXZ10</td><td></td></tr><tr><td>Eye lubricant</td><td>Bayer Vital GmbH</td><td>2182442</td><td></td></tr><tr><td>Xylazine</td><td>Bayer Vital GmbH</td><td>1320422</td><td></td></tr><tr><td>Ketamine</td><td>Serumwerke Bernburg</td><td>7005294</td><td></td></tr><tr><td>Tramadol</td><td>Gr&amp;uuml;nenthal GmbH</td><td>2256241</td><td></td></tr><tr><td>Disinfection solution (SoftaseptN)</td><td>Braun Melsungen AG</td><td>8505018</td><td></td></tr><tr><td>CD-1 mice</td><td>Charles River</td><td>22</td><td></td></tr></tbody></table>

an intramedullary locking nail for standardized fixation of femur osteotomies to analyze normal and defective bone healing in mice

Kemik iyileşmesi modelleri klinik kırık tedavisi için yeni tedavi stratejilerinin geliştirilmesine önem taşımaktadır. Bundan başka, fare modelleri, travma araştırmalarında kullanılan daha yaygın hale gelmektedir. Onlar kemik iyileşmesi çok farklılaşmış sürecin arkasındaki moleküler mekanizmaların analizi için mutant ve antikorların çok sayıda sunuyoruz. biyomekanik ortamı kontrol etmek için, standart ve iyi karakterize osteosentez teknikleri farelerde zorunludur. Burada, farelerde açık femur osteotomileri stabilize tasarım ve intramedüller çivi kullanımı ile ilgili rapor. Tıbbi kalite paslanmaz çelikten yapılmış çivi, yüksek eksenel ve dönme sertliği sağlar. ayrıca implant tanımlanan, sürekli osteotomi boşluğu yaratılması 0.00 mm 2.00 mm boyutları verir. 0.00 mm ve 0.25 mm boşluk boyutları ile femur osteotomi tırnak stabilizasyonu kilitleme intramedüller enkondral ve intramembranöz ossificat yoluyla yeterli kemik iyileşmesi sonucuiyon. atrofik olmayan birliğe 2.00 mm&#39;lik sonuçlarının bir boşluk boyutu ile femur osteotomi istikrar. Bu durumda, kemik iliği kilitleme tırnağı iyileşmesi ve olmayan iyileşme modellerinde kullanılabilir. Açık olan diğer kemik iyileşmesi modelleri ile karşılaştırıldığında tırnak kullanımının bir başka avantajı yeterince kemik bütünleşme sürecini incelemek için kemik yerine kullanılan ve iskeleler düzeltmek için imkanı vardır. intramedüller çivi kullanımı bir dezavantajı kapalı modellere kıyasla tüm açık prosedürlere doğasında daha invazif cerrahi işlemdir. Bir başka dezavantaj, ekstra-medular sabitleme işlemleri ile karşılaştırıldığında tüm intramedüller sabitleyici özgü intramedüler kavite bazı hasar indüksiyonu, olabilir.

cerrahi işlem için genel zaman yara kapanmadan yaklaşık 30 dakika cilt kesisi oldu. sağlanan cerrahi implantları kullanarak, cerrahi bir stereo-mikroskop olmadan yapılabilir. Ameliyat sonrası hayvanlar günlük olarak izlenmiştir. Hayvanların hiçbiri bu süre sonrası ağrı (seslendirme, huzursuzluk, hareketlilik olmaması, damat başarısızlık, anormal duruş ya da çevresi normal ilgi eksikliği) kanıt gösterdi çünkü post-operatif analjezi 3 gün sonra sona erdirildi...

Watch this Scientific Journal Video about An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice at JoVE.com

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

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 ...

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Research

JoVE Journal

Medicine

13.3K Görüntüleme.  Saarland University. 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.

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

Creating Rigidly Stabilized Fractures for Assessing Intramembranous Ossification, Distraction Osteogenesis, or Healing of Critical Sized Defects

Assessing modes of skeletal repair is essential for developing therapies to be used clinically to treat fractures. Mechanical stability plays a large role in healing of bone injuries. In the worst-case scenario mechanical instability can lead to delayed or non-union in humans. However, motion can also stimulate the healing process. In fractures that have motion cartilage forms to stabilize the fracture bone ends, and this cartilage is gradually replaced by bone through recapitulation of the developmental process of endochondral ossification. In contrast, if a bone fracture is rigidly stabilized bone forms directly via intramembranous ossification. Clinically, both endochondral and intramembranous ossification occur simultaneously. To effectively replicate this process investigators insert a pin into the medullary canal of the fractured bone as described by Bonnarens4. This experimental method provides excellent lateral stability while allowing rotational instability to persist. However, our understanding of the mechanisms that regulate these two distinct processes can also be enhanced by experimentally isolating each of these processes. We have developed a stabilization protocol that provides rotational and lateral stabilization. In this model, intramembranous ossification is the only mode of healing that is observed, and healing parameters can be compared among different strains of genetically modified mice 5-7, after application of bioactive molecules 8,9, after altering physiological parameters of healing 10, after modifying the amount or time of stabilization 11, after distraction osteogenesis 12, after creation of a non-union 13, or after creation of a critical sized defect. Here, we illustrate how to apply the modified Ilizarov fixators for studying tibial fracture healing and distraction osteogenesis in mice.

This article describes a method for stabilizing long bone fractures that is based on the application of modified Ilizarov external fixators 1-3. After application of the fixators and creation of the bone injury, healing can be assessed, distraction osteogenesis can be performed, or non-union or critical sized defect can be created and used to study therapeutic interventions.

Distraksiyon Osteogenezi, ya da Eleştirel Ölçekli Kusurların Şifa, Intramembranous kemikleşme değerlendirilmesi için Kademe Stabilize Kırıklar oluşturma

Assessing modes of skeletal repair is essential for developing therapies to be used clinically to treat fractures. Mechanical stability plays a large role ...

Tıp

A Mini-Invasive Internal Fixation Technique for Studying Immobilization-Induced Knee Flexion Contracture in Rats

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to restrict knee joint mobility is a widely accepted model to generate experimental contracture. However, implanting application will inevitably cause surgical trauma to the animals. Aiming to develop a less invasive approach, we combined a muscle-gap separation modus with a previously reported mini-incision skill during the surgical procedure: Two mini skin incisions were made on the lateral thigh and leg, followed by performing muscle-gap separation to expose the bone surface. The rat knee joint was gradually immobilized by a preconstructed internal fixation at approximately 135&#176; knee flexion without interfering essential nerves or blood vessels. As expected, this simple technique permits rapid postoperative rehabilitation in animals. The correct position of the internal fixation was confirmed by an x-ray or micro-CT scanning analysis. The range of motion was significantly restricted in the immobilized knee joint than that observed in the contralateral knee joint demonstrating the effectiveness of this model. Besides, histological analysis revealed the development of fibrous deposition and adhesion in the posterior-superior knee joint capsule over time. Thus, this mini-invasive model may be suitable for mimicking the development of immobilized knee joint contracture.

Here, we present a protocol to describe a minimally invasive technique for knee joint immobilization in a rat model. This reproducible protocol, basing on muscle-gap separation modus and the mini-incision skill, is suitable for studying the underlying molecular mechanism of acquired joint contracture.

Rats Immobilizasyon kaynaklı diz fleksiyon kontraktürü eğitim için mini-Invaziv Iç Fikmasyon tekniği

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to ...

Gelişim Biyolojisi

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 ...

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 ...

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

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies that promote efficient bone healing are non-existent and in high demand. Effective and reproducible animal models of fractures healing are needed to understand the complex biological processes associated with bone regeneration. Many animal models of fracture healing have been generated over the years; however, murine fracture models have recently emerged as powerful tools to study bone healing. A variety of open and closed models have been developed, but the closed femoral fracture model stands out as a simple method for generating rapid and reproducible results in a physiologically relevant manner. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice and facilitate a post-fracture stabilization of the femur by inserting an intramedullary steel rod. Although devices such as a nail or a screw offer greater axial and rotational stability, the use of an intramedullary rod provides a sufficient stabilization for consistent healing outcomes without producing new defects in the bone tissue or damaging nearby soft tissue. Radiographic imaging is used to monitor the progression of callus formation, bony union, and subsequent remodeling of the bony callus. Bone healing outcomes are typically associated with the strength of the healed bone and measured with torsional testing. Still, understanding the early cellular and molecular events associated with fracture repair is critical in the study of bone tissue regeneration. The closed femoral fracture model in mice with intramedullary fixation serves as an attractive platform to study bone fracture healing and evaluate therapeutic strategies to accelerate healing.

The murine closed femoral fracture model is a powerful platform to study fracture healing and novel therapeutic strategies to accelerate bone regeneration. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice using an intramedullary steel rod to stabilize the femur.

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 ...

Fracture Apparatus Design and Protocol Optimization for Closed-stabilized Fractures in Rodents

The reliable generation of consistent stabilized fractures in animal models is essential for understanding the biology of bone regeneration and developing therapeutics and devices. However, available injury models are plagued by inconsistency resulting in wasted animals and resources and imperfect data. To address this problem of fracture heterogeneity, the purpose of the method described herein is to optimize fracture generation parameters specific to each animal and yield a consistent fracture location and pattern. This protocol accounts for variations in bone size and morphology that may exist between mouse strains and can be adapted to generate consistent fractures in other species, such as rat. Additionally, a cost-effective, adjustable fracture apparatus is described. Compared to current stabilized fracture techniques, the optimization protocol and new fracture apparatus demonstrate increased consistency in stabilized fracture patterns and locations. Using optimized parameters specific to the sample type, the described protocol increases the precision of induced traumas, minimizing the fracture heterogeneity typically observed in closed-fracture generation procedures.

The goal of the protocol is to optimize the fracture generation parameters to yield consistent fractures. This protocol accounts for the variations in bone size and morphology that may exist between animals. Additionally, a cost-effective, adjustable fracture apparatus is described.

Kırık cihazları tasarımı ve kapalı-sağlamlık kırıklar Rodents için protokol optimizasyon

The reliable generation of consistent stabilized fractures in animal models is essential for understanding the biology of bone regeneration and developing ...

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 ...