Bu çalışma NIAMS gelen R01-AR053645 tarafından finanse edildi.

Tüm Prosedürler UCSF Kurumsal Hayvan Bakımı tarafından onaylanmış ve Komite kullanın ve ulusal kurallara uygun yapıldı. 1. Cerrahi öncesinde Fiksatörleri hazırlanması <ol><li> Stabilize kırık oluşturmadan önce, bir özel-tasarım eksternal fiksatör cihazı monte etmek gerekiyor. Özel tasarlanmış eksternal fiksatör kare üç paslanmaz çelik # 0/56 dişli çubuklar, 8 # 2/56 altıgen somun ve cıvata 17 eşleştirme (Küçük Parça Inc, Miami, FL 3) ile stabilize iki alüminyum halkalar oluşuyordu. </li><li> İki alüminyum halkaları kırık başına ihtiyaç vardır. Bir halka # 2/56 altıgen somunlar (Küçük Parça Inc, Miami, FL) ve eşleme cıvataları uygun deliklere 4 birleştirmek için, bu gevşek kalmalıdır. Bu proksimal halkası olacaktır. Distal halka 3 somun ve cıvata birleştirmek için tekrar tam sıkmayın. </li></ol> 2. Anestezi, Kırılma Yaratılış ve Fiksatör Uygulaması <ol><li> Fare (yaş, cinsiyet ve vücutkilo 50mg/ml Ketamin, 0.5 intraperitoneal enjeksiyonu ile anestezi) kullanıcı tarafından edilir belirlenir mg / ml medetomidin (0.03 ml / fare). </li><li> Farelerin gözlerine göz yağlayıcı işlem sırasında gözlerini korumak için kendi başının üzerinden bant parçası koyun. </li><li> Hayvan prosedür kalanı için bir ısıtma yüzeyi üzerine yerleştirilir. </li><li> Mıhlamak dört 0,25 mm steril böcek iğneli (Anticorro: Güzel Bilim Aracı, Foster City, CA) kullanılarak sol tibia proksimal ve distal metafiz bir Dremel Aracı pin ekleyerek ve kemik yoluyla delerek. </li><li> Pimler yönelimli kaval kemiğinin uzun eksenine dik ve birbirine 90 ° vardır. En az 10 proksimal ve distal pin arasında mm. </li><li> Pimleri yönünde dönük fındık ile proksimal pin üzeri ilk halkası yerleştirin. Alüminyum halka içinde merkezi olarak konumlandırılmış tibia tutarak halka pimlerini emniyete alın. </li><li> Di aşağıda ikinci halka konumlandırmakstal pimleri ve daha önce olduğu gibi altıgen somunları kullanarak halka pimleri sabitleyin. Distal fiksatöre dördüncü somun-cıvata kombinasyonu ekleyin. </li><li> Uygun bir cihaz ile bükme üç nokta ile transvers kırık oluşturun. Biz Bonnarens ve Einhorn 4 yerçekimi sistemi kullanınız. Bloklar arasında tibia posterolateral yüzeyi dinlendirin, açılan kol boyunca önceden belirlenmiş bir yüksekliğe ağırlığı artırmak ve ağırlık düşmesi temas künt kırık üretmek için tibianın anterolateral yönü böylece. Boy kilo tibianın sadece tek bir kırık sağlamak için fare her bir suş için belirlenecek ihtiyaçlar dışında bırakılmıştır. </li><li> Kırılma X-ray ile teyit edilir. </li><li> Kırık onayladıktan sonra, üç paslanmaz çelik # 0/56 dişli çubuklar halkaları eklenir. Distal ucundan başlayarak çubuk takın ve çubuk sabitlemek üzere distal halkası üzerine bir somun geçiriniz. Sonra çubuk üzerine başka bir somun koymak, proksimal halka bu n üstüne oturacağımut. Iki halka eşit aralıklarla yapılacak ve pimleri zorlayarak değil emin olun. Üçüncü bir somunu ile çubukların proksimal halka sabitleyin. </li><li> Analjezi Buprenorfinin Subkutan enjeksiyonu (1.0 mg / kg) ameliyat sonrası hemen verilir ve gerektiği gibi hayvanlara acı ve verilen buprenorfin için izlenir. </li><li> Anestezi Atipamezole idare tarafından iptal edilir ve bunlar duyarlı ve yürüyebilen kadar hayvanlar izlenir. </li></ol> 3. Distraksiyon Osteogenezi (ayrıca bkz: 12,13) Karşılamak için bu yordamı değiştirmek için distraksiyon osteogenezisi basittir. Halkaları dişli çubuklarla konumda tutulur ve yerine çubuklar tutan fındık çevirerek halkaları ayrı hareket ettirilebilir. <ol><li> Gecikme: Bu model kullanılarak standart distraksiyon osteogenezisi için fiksatör tibia üzerine yerleştirilir ve 5 günlük bir gecikme süre yalnız bıraktı. </li><li> Distraksiyon: Distraksiyon osteogenesis dengeleyici çubuk ¼ dönüş (0,25 mm), her gün 7 gün üzerindeki somun çevrilerek gerçekleştirilebilir. </li><li> Olgunlaşma: Kemikli köprü 10 gün gözlenir ve tam konsolidasyon distraksiyon tamamlanmasından sonra 27 gün görülmektedir. </li></ol> 4. Bir Kritik Ölçekli Hata oluşturulması Daha önce aşağıdaki değişiklik ile tarif edildiği gibi kritik bir büyüklükte kusurun oluşturmak için dış sabitleyicilerin uygulanır. <ol><li> Prosedürü başlamadan önce cilt% 70 etanol ile temizlenir. </li><li> Fiksatörler defekt oluşturarak önce tamamen uygulanır. </li><li> Fare daha sonra medial tibia görselleştirmek için diseksiyon mikroskobu altında koyun, ve bir kesi (5-7 mm) bir neşter bir # 10 bıçak ile yapılır. </li><li> Kas hafifçe transeksiyon ve tibianın orta diafiz maruz ayrılır. </li><li> Kemiğin bir 3 mm segment daha sonra orta diyafiz kullanılarak makas kaldırılır. T kullanmadiye makas işaret, kemik yavaş yavaş snipped ve kemik parçaları gerekirse forseps ile çıkarılır. Bakım büyük bir mekanik kuvvet ile tibia yerinden önlemek için makas uçları ile alınmalıdır. </li><li> Bu iyileşmesine katkıda olabilir diye bir kemik parçası, orada olduğundan emin olmak için cerrahi sitesini inceleyin. </li><li> Tibia künt ucu çevresindeki kas değiştirin. </li><li> 6-0 Poliamin Monofilament 2-3 sütürler ile cilt kesisi kapatın. </li><li> Anestezi tersine çevrilir ve hayvanlar için izlenir ağrısı ve daha önce tarif edildiği gibi analjezikler verilmiştir. </li></ol> 5.. Temsilcisi Sonuçlar Doğru uygulandığında, eksternal fiksatör mükemmel bir azalma (Şekil 1, 2) kapalı tibia kırığı daha sert istikrar sağlamak. Ancak, bazı durumlarda yetersiz azalma (kemik uçlarının veya birden fazla kırıklar arasında belirgin ve büyük bir boşluk (Şekil 3 meydana </strong&gt;), Ve bu farelerde analizlerin dışında tutulmuştur. Kırıkları, öncelikle intramembranous kemikleşme (Şekil 4) aracılığıyla iyileşmesi, bu yöntem kullanılarak stabilize. Kırık stabilize değilse Bunun aksine, geniş bir kıkırdak kallus kırık boşluğu (Şekil 5) içinde oluşturulmuş olup, bu intramembranous kemikleşme işlemi boyunca kemik ile değiştirilir. <img alt="Şekil 1" src="/files/ftp_upload/3552/3552fig1.jpg" /> Şekil 1. Tibia kırığı sabitlemek için kullanılan bir harici fiksasyon cihazı gösteren radyografileri. Grafisinde iyi hizalanmış kemik segmentleri (ok) gösteren kırık sonrası alınmıştır. <img alt="Şekil 2" src="/files/ftp_upload/3552/3552fig2.jpg" /> Şekil 2. Fiksatör sonra bir fare resmi uygulanmıştır. <img alt="Şekil 3" src="/files/ftp_upload/3552/3552fig3.jpg" /> Şekil 3. Radyografi f sonra alınanracture hizalanmış ve parçalanmış kemik segmentleri (ok) göstermektedir. <img alt="Şekil 4" src="/files/ftp_upload/3552/3552fig4.jpg" /> Şekil 4. Stabilize kırık intramembranous ossifikasyon ile iyileşir. Stabilize kırık trikrom kırık bölgesinde bazı yeni kemik (b) gösterir. Ölçek çubuğu = 500 mikron. <img alt="Şekil 5," src="/files/ftp_upload/3552/3552fig5.jpg" /> Şekil 5. Sigara stabilize kırık yon ile iyileşir. Non-stabilize kırık trikrom (b) kırık yerinde kıkırdak (c) ve kemik gösterir. Ölçek çubuğu = 500 mikron.

<ol>
	<li>Ilizarov, G. A., Lediaev, V. I., Shitin, V. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+course+of+compact+bone+reparative+regeneration+in+distraction+osteosynthesis+under+different+conditions+of+bone+fragment+fixation+(experimental+study).">The course of compact bone reparative regeneration in distraction osteosynthesis under different conditions of bone fragment fixation (experimental study).</a> Eksperimentalnaia Khirurgiia i Anesteziologiia. 14, 3-12 (1969).</li><li>Ilizarov, G. A., Deviatov, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+elongation+of+the+leg.">Surgical elongation of the leg.</a> Ortopediia Travmatologiia i Protezirovanie. 32, (1971).</li><li>Thompson, Z., Miclau, T., Hu, D., Helms, J. 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Biz ifşa hiçbir şey yok.

Bones mekanik stabilite (: 14 Yorumlar) bağlı olarak iki farklı modaliteleri ile iyileşir. Kırık kemiğin iki ucu köprü kemik ile değiştirilir kırık boşluğu kararsız büyük bir kıkırdak şablon formları ayrıldığımda. Proksimal ve distal mola için, kemik, periost ve endosteum içinde intramembranous ossifikasyon doğrudan oluşturur. Bunun aksine, kararlı bir kırık iyileşmesi intramembranous kemikleşmesi 3 vasıtasıyla yalnızca oluşur. Ancak, bu iki süreç arasındak...

<table border="1"><tbody><tr><td> Reaktif Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> 0.25mm böcek pin </td><td> Güzel Bilim Aracı </td><td> 26000-25 </td><td> Blacked Eloksal Çelik, 0.25mm çubuk çapı, 4 cm uzunluk </td></tr><tr><td> Paslanmaz Çelik Hex Nut </td><td> Küçük Parçalar </td><td> # 2.56 </td><td> 1/8 &quot;uzunluğu, inç başına 56 Konu </td></tr><tr><td> Paslanmaz Çelik Hex Nut </td><td> Küçük Parçalar </td><td> # 0-80 </td><td> 1/8 &quot;uzunluğu, inç başına 80 Konu </td></tr><tr><td> Paslanmaz Çelik Makina Vida </td><td> Küçük Parçalar </td><td> # 0-80 </td><td> 1/8 &quot;uzunluğu, inç başına 80 Konu </td></tr><tr><td> Paslanmaz Çelik Makina Dişli Çubuk Kesme </td><td> Küçük Parçalar </td><td> # 0-80 </td><td> 6 &quot;uzunluğunda, inç başına 80 Konu </td></tr><tr><td> 18-8 Paslanmaz Çelik Baş Makinası Screw </td><td> McMaster-Carr </td><td></td><td> 2-56 konu, 3/6 &quot;uzunluk </td></tr><tr><td> Dış Fiksasyon Cihazı </td><td> Atölye </td><td> Özel tasarlanmış </td><td></td></tr></tbody></table>

creating rigidly stabilized fractures for assessing intramembranous ossification, distraction osteogenesis, or healing of critical sized defects

Iskelet tamir modları değerlendirmek kırıkların tedavisi için klinik olarak kullanılmak üzere terapiler geliştirmek için gereklidir. Mekanik stabilite kemik yaralanmaları iyileşmesinde büyük bir rol oynar. En kötü durum senaryosunda mekanik istikrarsızlık gecikmiş veya sendika üyesi olmayan insanlara yol açabilir. Ancak, hareket de iyileşme sürecini uyarabilir. Kırık kemik uçlarının stabilize etmek için hareket kıkırdak formları var ve bu kıkırdak yavaş yavaş yon gelişim sürecinin rekapitülasyon yoluyla kemik ile değiştirilir kırıklarda. Bir kemik kırığı katı stabilize Buna zıt olarak, kemik intramembranous kemikleşme yoluyla doğrudan oluşturur. Klinik olarak, hem endokondral ve intramembranous ossifikasyon aynı anda gerçekleşir. Etkili bir şekilde bu süreci araştırmacılar Bonnarens 4 tarafından açıklandığı gibi kırık kemiğin medüller kanal içine bir iğne eklemek çoğaltmak için. Rotatio sağlarken Bu deneysel yöntem mükemmel yanal stabilite sağlarnal istikrarsızlık devam etmek. Ancak, bu iki farklı süreçlerini düzenleyen mekanizmaların anlaşılmasında deneysel olarak da bu süreçlerin her biri izole tarafından geliştirilmiş olabilir. Biz rotasyonel ve yanal denge sağlar istikrar protokolü geliştirdi. Bu modelde, intramembranous ossifikasyon görülür iyileşme tek mod ve şifa parametrelerin iyileşmesi 10 fizyolojik parametreleri değiştirmeden sonra biyoaktif moleküller 8,9 uygulamasından sonra 5-7 genetiği değiştirilmiş farelerde farklı suşları,,, arasında mukayese edilebilir distraksiyon osteogenezisi 12 sonrasında, stabilizasyon 11 miktarı veya zaman değiştirdikten sonra, bir sendika üyesi olmayan 13 oluşturulduktan sonra, ya da kritik bir boyutta defekt oluşturulduktan sonra. Burada, farelerde tibial kırık iyileşmesi ve distraksiyon osteogenezisi çalışmak için modifiye İlizarov fiksatör nasıl uygulanacağını göstermektedir.

Watch this Scientific Journal Video about Creating Rigidly Stabilized Fractures for Assessing Intramembranous Ossification, Distraction Osteogenesis, or Healing of Critical Sized Defects at JoVE.com

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

Bu makale, modifiye İlizarov yimi uygulamaya dayanır uzun kemik kırığı stabilize edilmesi için bir yöntemi açıklar<sup&gt; 1-3</sup&gt;. Sabitleyiciler ve kemiğin zarar oluşturulması uygulamasından sonra, iyileştirici değerlendirilebilir, distraksiyon osteogenezisi yapılabilir veya birleşik-olmayan veya kritik ölçekli kusurun oluşturulur ve tedavi edici müdahaleler incelemek için kullanılabilir.

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 ...

creating-rigidly-stabilized-fractures-for-assessing-intramembranous

Research

JoVE Journal

Medicine

17.3K Görüntüleme.  University of California, San Francisco. Bu makale, modifiye İlizarov yimi uygulamaya dayanır uzun kemik kırığı stabilize edilmesi için bir yöntemi açıklar 1-3. Sabitleyiciler ve kemiğin zarar oluşturulması uygulamasından sonra, iyileştirici değerlendirilebilir, distraksiyon osteogenezisi yapılabilir veya birleşik-olmayan veya kritik ölçekli kusurun oluşturulur ve tedavi edici müdahaleler incelemek için kullanılabilir.

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

Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. Adipose-derived stromal cells (ASCs) have proven to be an abundant source of multipotent stem cells capable of undergoing osteogenic, chondrogenic, adipogenic, and myogenic differentiation. Many studies have explored the osteogenic potential of these cells in vivo with the use of various scaffolding biomaterials for cellular delivery. It has been demonstrated that by utilizing an osteoconductive, hydroxyapatite-coated poly(lactic-co-glycolic acid) (HA-PLGA) scaffold seeded with ASCs, a critical-sized calvarial defect, a defect that is defined by its inability to undergo spontaneous healing over the lifetime of the animal, can be effectively show robust osseous regeneration. This in vivo model demonstrates the basis of translational approaches aimed to regenerate the bone tissue - the cellular component and biological matrix. This method serves as a model for the ultimate clinical application of a progenitor cell towards the repair of a specific tissue defect.

This protocol describes the isolation of adipose-derived stromal cells from lipoaspirate and the creation of a 4 mm critical-sized calvarial defect to evaluate skeletal regeneration.

Lipoaspirate itibaren hasat Yağ kaynaklı stromal hücreler kullanarak Kritik ölçekli Kalvaryum Defekt Model tamiri

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. ...

Tıp

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. ...

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

One desirable endpoint of general anesthesia is the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is less straightforward than it is in human patients. A widely used behavioral surrogate for hypnosis in rodents is the loss of righting reflex (LORR), or the point at which the animal no longer responds to their innate instinct to avoid the vulnerability of dorsal recumbency. We have developed a system to assess LORR in 24 mice simultaneously while carefully controlling for potential confounds, including temperature fluctuations and varying gas flows. These chambers permit reliable assessment of anesthetic sensitivity as measured by latency to return of the righting reflex (RORR) following a fixed anesthetic exposure. Alternatively, using stepwise increases (or decreases) in anesthetic concentration, the chambers also enable determination of a population's sensitivity to induction (or emergence) as measured by EC50 and Hill slope. Finally, the controlled environmental chambers described here can be adapted for a variety of alternative uses, including inhaled delivery of other drugs, toxicology studies, and simultaneous real-time monitoring of vital signs.

Loss of the righting reflex has long served as a standard behavioral surrogate for unconsciousness, also called hypnosis, in laboratory animals. Alterations in volatile anesthetic sensitivity caused by pharmacological interventions can be detected with a carefully controlled high-throughput assessment system, which may be adapted for delivery of any inhaled therapeutic.

Yerel ya da Sistemik Farmakolojik Müdahale sonrası Farelerin Uçucu Genel Anestezik Hassasiyet Değişiklikleri değerlendirilmesi

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve patient outcomes. We present our experience with a minimally invasive pterional approach for anterior circulation aneurysms performed in a major tertiary cerebrovascular center and compare the results with an aged matched dataset from the Nationwide Inpatient Sample (NIS). From August 2008 to December 2012, 22 elective aneurysm clippings on patients &#8804;55 years of age were performed by the same dual fellowship-trained cerebrovascular/endovascular neurosurgeon. One patient (4.5%) experienced transient post-operative complications. 18 of 22 patients returned for follow-up imaging and there were no recurrences through an average duration of 22 months. A search in the NIS database from 2008 to 2010, also for patients aged &#8804;55 years of age, yielded 1,341 hospitalizations for surgical clip ligation of unruptured cerebral aneurysms. Inpatient length of stay and hospital charges at our institution using the minimally invasive thumb-sized pterional technique were nearly half that of NIS (length of stay: 3.2 vs 5.7 days; hospital charges: $52,779 vs. $101,882). The minimally invasive thumb-sized pterional craniotomy allows good exposure of unruptured small and medium-sized supraclinoid anterior circulation aneurysms. Cerebrospinal fluid drainage from key subarachnoid cisterns and constant bimanual microsurgical techniques avoid the need for retractors which can cause contusions, localized venous infarctions, and post-operative cerebral edema at the retractor sites. Utilizing this set of techniques has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

Minimally invasive thumb-sized pterional craniotomy for aneurysm clipping has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

Minimal İnvazif Thumb-boyutlu pterional Kraniyotomi Kanamamış Anterior Sirkülasyon Anevrizmalarının Cerrahi Klip Ligasyonu için

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve ...

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 ...

Direct Mouse Trauma/Burn Model of Heterotopic Ossification

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical procedures. The majority of animal models used to study HO rely on the application of exogenous substances, such as bone morphogenetic protein (BMP), exogenous cell constructs, or genetic mutations in BMP signaling. While these models are useful they do not accurately reproduce the inflammatory states that cause the majority of cases of HO. Here we describe a burn/tenotomy model in mice that reliably produces focused HO. This protocol involves creating a 30% total body surface area partial thickness contact burn on the dorsal skin as well as division of the Achilles tendon at its midpoint. Relying solely on traumatic injury to induce HO at a predictable location allows for time-course study of endochondral heterotopic bone formation from intrinsic physiologic processes and environment only. This method could prove instrumental in understanding the inflammatory and osteogenic pathways involved in trauma-induced HO. Furthermore, because HO develops in a predictable location and time-course in this model, it allows for research to improve early imaging strategies and treatment modalities to prevent HO formation.

An Achilles tenotomy and burn injury model of heterotopic ossification allows for the reliable study of trauma induced ectopic bone formation without the application of exogenous factors.

Doğrudan Fare Travma / Heterotopik Kemikleşme Model Yanık

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical ...

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 ...

Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal abnormalities. In addition, pathological myopia is a common irreversible cause of visual impairment and blindness1-3. Our study demonstrates the effect of scleral crosslinking using riboflavin and ultraviolet-A radiation on the development of axial myopia in a rabbit model. The axial length of the eyeball was measured by A-scan ultrasound in New Zealand white rabbits aged 13 days (male and female). The eye then underwent 360&#176; conjunctival peritomy with scleral crosslinking, followed by tarsorrhaphy. Axial elongation was induced in 13 day-old New Zealand rabbits by suturing their right eye eyelids (tarsorrhaphy). The eyes were divided into quadrants, and every quadrant had two scleral irradiation zones, each with an area of 0.2 cm&#178; and a radius of 4 mm. Crosslinking was performed by dropping 0.1% dextran-free riboflavin-5-phosphate onto the irradiation zones 20 sec before ultraviolet-A irradiation and every 20 sec during the 200 sec irradiation time. UVA radiation (370 nm) was applied perpendicular to the sclera at 57 mW/cm&#178; (total UVA light dose, 57 J/cm&#178;). Tarsorrhaphies were removed on day 55, followed by repeated axial length measurements. This study demonstrates that scleral crosslinking with riboflavin and ultraviolet-A radiation effectively prevents occlusion-induced axial elongation in a rabbit model.

We demonstrate the effect of scleral crosslinking with riboflavin and UVA on an axial elongation rabbit eye. Axial elongation was induced in 13 day-old New Zealand rabbits (male and female) by suturing their right eye eyelids (tarsorrhaphy).

Skleral bir Tavşan Modelinde Aksiyel Miyopide Önleme Riboflavin ve Ultraviyole-A Radyasyon Kullanımı Çapraz bağlama

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal ...

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 ...