We thank members of the Mariani lab for critical reading of the manuscript. Our funding sources were: the Baxter Medical Scholar Research Fellowship (to M.K.S.), USC undergraduate fellowships and the Provost, Dean Joan M. Schaeffer, and Rose Hills Fellowships, (to M.K.S.). We also acknowledge a CIRM BRIDGES fellowship through Pasadena City College (to T.T.T). and the James H. Zumberge Research and Innovation Fund, the USC Regenerative Medicine Initiative, and the NIAMS NIH under Award Number R21AR064462 (to F.V.M).

NOT: Tüm işlemler Southern California Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) tarafından onaylanmış bir hayvan protokolüne uygun bulunmaktadır. Cerrahi için 1. Hazırlık <ol><li> Gerekli cerrahi aletleri ve malzemeleri yerleştirin (1 öğe sterilize - Malzeme listesinde 18) ve bir mikroskop ve bir başlık ya da bir aşağı doğru masaya bir fiber optik trans-aydınlatıcı düzenlemek. </li><li> Anestezi ikna etmek için% 4 izofluran ile bir indüksiyon odasında (3 aylık - - 30 gr, 1 20) cinsel olgun fare yerleştirin. </li><li> Bir pençe ve / veya kuyruk tutam testi ile anestezi onaylayın. </li><li> Ameliyat sırasında göz kuruluğu önlemek için farenin gözlerine merhem sürün. </li><li> Mikroskop sahneye fareyi hareket ettirin. Bakım hortum burun konisi içine fare koyun ve yanal yerleştirin. Vücut ısısını korumak amacıyla bir hayvana bitişik ısıtıcı bir torba sağlar. DEĞİLE:. Fare Pozisyon kişisel tercihi ve / veya cerrahın ellilik bağlıdır, sol veya sağ ya da yan gayet iyi. </li><li> Bakım için% 3, ve solunum hızı izle - 2 izofluran ayarlayın. </li><li> Bir 25 G iğne ile ameliyat sonrası ağrı için buprenorfın (0.05mg / kg) deri altından (bacağın lateral yan) enjekte edilir. bacak normal bir tepki olduğu seğirme olabilir. </li></ol> 2. Açılış Kesi <ol><li> Palpasyonla istenen kaburga Algılama ve tıraş ile saç alanını temizleyin. NOT: kaburga 1-7 akciğerlere yakın ve 11-13 oldukça kısa olduğundan kaburga 8-10 tavsiye edilir. Kaburga Sayma 1 numara olarak en rostral kaburga ile başlamalıdır. </li><li> (3x genellikle hayvan kullanım protokolleri tarafından tavsiye edilir), povidon-iyot ve izopropanol (% 70) sürüntü alternatif alanı hazırlayın. </li><li> Doğrudan yukarıda, deri yoluyla 2 cm kesi kesin ve orta ölçekli mikrocerrahi makas ile istenilen kaburga paralel. INCISaltta yatan kas ve yağ katmanları aracılığıyla e. </li><li> Kesi boyutu en aza indirirken, cerrahi alan ortaya çıkarmak için bir sarıcıya 3 katmanları (deri, kas, yağ) yerleştirin. </li></ol> 3. kaburga eksize <ol><li> Bir 5,0 mm neşter ile kaburga istenilen bölümünü örten interkostal kaslar kesti. Kaburga gibi son derece kavisli olmadığı bu gibi chondrocostal eklem pozisyon 5 mm proksimal etrafında bu kesi olduğunu. Dikkatle ince uçlu forseps ile kemik kas ayrı. </li><li> Hayvanda periost koruyarak bir reseksiyona oluşturmak için, bir 5.0 mm bir bisturi ile şeridin uzunluğu boyunca, periost kesilmiştir. Dikkatle ince uçlu forseps ile yanal altta yatan kemik periost ayrı. Periost çok hassas ve bir jelatinimsi kıvama sahip olduğu, dikkatli devam. </li><li> Sonraki ince mikro-makas ile bir ucunda kemik yoluyla bir kesitini yapmak. Gerekirse, ile rezeksiyon ölçmekmikroskop veya cetvel türü göstergesi bir reticle. Sonra dikkatlice periost dışarı kemik kaldırın ve diğer ucunu kesti. NOT: Dikkatli devam edin. Bu plevral membran gözyaşı ve bir pnömotoraks neden aşağıda kaburga eksize olmadan yukarı çekerek gibi en hassas adımdır. Plevra zarı yırtılmış ise göğüs boşluğunun iç görünür hale gelecektir bu kolaylıkla belli olacak. </li><li> Kanamayı durdurmak için 5 sn - kanama kemik kesme sonucu oluşursa, 4 için pamuk uçlu çubuk ile kesik sonunda basınç uygulayın. NOT: Nadiren kanama devam ettiğini, bunu durdurmak ve hayvana kurtarma tehlikeye gibi önemli kan kaybı euthanize gerekli olabilir (sıvı replasmanı olmadan, güvenle kaybolabilir maksimum kan hacmi, toplam kan hacminin% 10 ya da 7,7 -.) 200 ul 9 - 8 ul / g, 25 g bir fare için, bu yaklaşık 180 ile eşdeğerdir. </li><li> Hemen içine kaldırıldı kaburga yerleştirinGelecekteki analiz için% 4 PFA. NOT: kemik ve periost hem rezeksiyonu yapılacak ise, periost kesi ve ayrılık (adım 3.2) ihmal. Periost çok sıkı plevral zara bağlı olduğundan yırtılmasını önlemek için forseps ile uzak plevral membran kaburga alay, dikkatli devam. </li></ol> 4. kesi Kapanış <ol><li> 9-0 sütür (2 sütür genellikle yeterlidir) ile kalan periost kovanın üstünden interkostal kasları sütür. Cerrahi konumu göstergeleri olarak hareket etmek, doğrudan kaburga kesim uçları yukarıda sütür yerleştirin. </li><li> Ekartörünü çıkarın. (- 4 sütür genellikle yeterli 3) 9-0 sütür ile örten kas ve yağ sütür. Kas ve / veya yağ, özellikle kalın tabakalar farelerde, her biri ayrı ayrı bir tabaka dikiş (örneğin, kas tabaka için dikiş 1 tabakasının yağ tabakası için dikiş 1 kat). </li><li> Kapat(- 5 sütür genellikle yeterlidir 4) 7-0 dikiş ile cilt. </li><li> Büyük forseps ile birlikte kenarları kısma, dikiş yapıştırıcı ile kesi sabitleyin. </li><li> Yavaş yavaş ilk birkaç dakika için% 1 ayarlayarak kapalı izofluran fare vazgeçirmek ve sonra kapatın. </li><li> Bir ısı lambası altında fare yerleştirin ve bilinç, 5 sağlanıncaya kadar bırakın - 10 dakika. Bilinç kazanmış sonra ve iyileşme dönemi boyunca, fare taşımak ve normal ambulate ve sıkıntı emaresi göstermelidir. </li></ol> 5. Kurtarma ve Analiz <ol><li> 0.5 mg bir oral jelatin formunda, buprenorfinin uygulama da dahil olmak üzere ameliyat sonrası ağrı idaresi sağlamak / 48 saat süre ile, her 12 saat kg. Oral uygulama göğüs bölgesinde ağrıya neden olabilir hayvan frenleyici gerektirmez. </li><li> Iyileşme döneminde yiyecek ve su ücretsiz erişim ile bir kafes içinde fare koruyun. Kesik kuruduktan sonra, kadın hayvanlar erkek ise birlikte muhafaza edilebilirs mücadele önlemek için izole kalmalıdır. </li><li> Iyileşme döneminden sonra, ötenazi onaylanmış prosedürü uygulayın. Tespit ve analiz için göğüs kafesini çıkarın. </li></ol>

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The authors have nothing to disclose

Önce bu protokolü öğrenirken, nerede zor olabilir başlangıç ​​kesi yerini belirlerken. Ancak, ötenazi fareler üzerinde uygulama cerrah ilk kesi yerleştirin ve rezeke istenen kaburga maruz nerede öğrenmenize yardımcı olur. Kadavra üzerinde çalışmak aynı zamanda veya periost olmadan kaburga kısmını kaldırmak için gerekli ince motor becerilerini geliştirir. Ayrıca, bu prosedüre yeni biri ince araçlar ve ince sütür manipüle zor bulabilir. Off bağlama sırasında, aşırı sütür çekerek...

Iskelet yaralanma, kronik osteoartrit zayıflatıcı ve rekonstrüktif cerrahi ile ilgili ciddi sorunlar, ekonomik verimliliği, aile refahı ve yaşam kalitesini etkileyebilir. Küçük molalar ve lezyonlar oldukça iyi iyileşmek mümkün olmakla birlikte, insanlar yapısını ve fonksiyonunu geri rekonstrüktif prosedürleri güvenmek gerekir, bu nedenle büyük kusurları tamir ve yeteneğine sahip değildir. İmar allojeneik veya Heterojenik greft, morcellized kemik, implante iskeleleri, ya da distraksiyon osteogenezisi içerebilir. Ne yazık ki, sadece bu tedaviler ancak tamir kemiğin orijinal gücü nadiren ulaşılır ile ilişkili inatçı morbidite faktörler vardır. Böylece, yeni klinik yaklaşımlara ihtiyaç vardır. Segmental kusurları tedavi etmek için yenilikçi yöntemler geliştirmek Tek yönlü büyük ölçekli onarım doğal olarak oluştuğu durumları incelemektir. Memeliler th sınırlı olarak kabul edilir ise Amfibi ünlü, iskelet elemanları yeniden olabiliryeteneğidir. Ancak, 20. yüzyılın başlarında bu yana, insan kaburga rejenerasyon birkaç rapor insanlar öylesine 1-4 sınırlı değildir düşündürmektedir yayınlanmıştır. Şu anda bu olay iyi çene, yüz ve kulak rekonstrüksiyonu için kaburga malzeme kullanımı, plastik cerrahlar tarafından bilinir, ama daha geniş 5 takdir edilmez. Daha ayrıntılı olarak bu onarım incelemek için, biz fareyi kullanarak bir cerrahi modeli geliştirdik. Bu protokolü kullanarak, araştırmacılar ilgili doğuştan faktörleri belirlemek ve diğer yerlerde iskelet şifa kolaylaştırmak için bu bilgileri kullanabilirsiniz. Iskelet onarım çalışmaları için bir model olarak kaburga kullanmanın birçok avantajı vardır. (Femur 6,7 rezeksiyonu kıyasla) İlk olarak, çevreleyen kaburgalar doğal fiksatör sağlar. Bu iç ve dış fiksatör morbidite riskini azaltır ve cerrahi prosedürü kolaylaştırır. Göğüs wa İkincisi, ince kas tabakalarıkolay erişim ve kalvaryal rezeksiyon- 8 kolaylık tahlil karşılaştırılabilir hale mükemmel bir görüş sağlamak ll. Üçüncüsü, intramembranöz kemikleşme, endokondral kemikleşme ile kaburga formu oluşması ve merkezi bir diafiz iki ucunda bulunan büyüme plaklarının de uzatma yoluyla boyu büyüdükçe kafataslarından aksine. Bu nedenle, kaburga tamir apendiküler iskeletin uzun kemiklerin onarımı daha karşılaştırılabilir olabilir. Ayrıca, femur göre, kaburga periost kalındır ve daha kolay bir şekilde manipüle edilebilir olduğunu bulmuşlardır. Böylece, periost okuyan veya hücre tedavileri, farmakolojik ajanlar test ve / veya doku iskelelerinin amacıyla kemik onarımı tahlil etmek isteyen araştırmacılar, bu cerrahi modeli yararlı bulabilirsiniz. Özet olarak, bu yiv rezeksiyon modeli yaygın olarak kullanılan bu tür bir model halen var olduğu memelilerde doğal büyük ölçekli kemik onarım çalışma içinde bir ortam sağlar.

<table><tbody><tr><td>Medium sized micro-dissection scissors (Vannas-T&amp;uuml;bingen Spring Scissors 5 mm)</td><td>Fine Science Tools</td><td>15003-08</td><td></td></tr><tr><td>Fine micro-dissection scissors (Vannas Spring Scissors - 2mm Cutting Edge)</td><td>Fine Science Tools</td><td>15000-04</td><td>curved tip is beneficial</td></tr><tr><td>Micro-scalpel 5.0 mm</td><td>Fine Science Tools</td><td>10315-12</td><td>other fine scalpels can be substituted</td></tr><tr><td>Dumont 55 forceps</td><td>Fine Science Tools</td><td>11295-51</td><td></td></tr><tr><td>Retractor&amp;nbsp;</td><td>Fine Science Tools</td><td>17004-05</td><td>adjustability is convenient</td></tr><tr><td>Micro-needle holders</td><td>Fine Science Tools</td><td>12060-01</td><td></td></tr><tr><td>9.0 nylon sutures (Ethilon), taper point best</td><td>Ethicon</td><td>2819G or similar</td><td>taper point best but reverse cutting is also good</td></tr><tr><td>7.0 prolene sutures (Prolene)</td><td>Ethicon</td><td>8700H or similar</td><td>6-0 can be used too, needle point can vary</td></tr><tr><td>Large forceps (Adson Forceps)</td><td>Fine Science Tools</td><td>11006-12</td><td>other brands are fine</td></tr><tr><td>Lubricant Eye Ointment (Akwa Tears)</td><td>Akorn</td><td>17478-062-35</td><td></td></tr><tr><td>Suture glue (GLUture Topical Tissue Adhesive)</td><td>Abbot</td><td>32046-01</td><td>has excellent working time</td></tr><tr><td>Shaver</td><td>Wahl</td><td>9918-6171 or similar</td><td></td></tr><tr><td>Clamp lamp</td><td>Zoo Med</td><td>LF-5</td><td></td></tr><tr><td>Infrared Bulb, 75W</td><td>Zoo Med</td><td>RS-75</td><td></td></tr><tr><td>RC&lt;sup&gt;2&lt;/sup&gt; Rodent Anesthesia System</td><td>VetEquip&amp;nbsp;</td><td>922100</td><td></td></tr><tr><td>IsoFlo (Isoflurane)</td><td>Abbot</td><td>05260-05</td><td></td></tr><tr><td>Buprenorphine (Buprenex)</td><td>Reckitt Benckiser</td><td>12496-0757-1</td><td></td></tr><tr><td>Betadine</td><td>Purdue Frederick</td><td>67618015017</td><td></td></tr><tr><td>Flavored Gelatin, raspberry</td><td>Jell-O</td><td>B000E1FYL0</td><td>made up firm, to the consistency of &#039;jigglers&#039;</td></tr></tbody></table>

a surgical procedure for resecting the mouse rib: a model for large-scale long bone repair

Bu protokol fare kaburga kullanan büyük ölçekli kemik onarımı için yeni bir model için araştırmacılar tanıttı. , çevredeki interkostal kaslar istenen kaburga açığa göğüs vücut duvarı açılış pnömotoraks neden olmadan kaburga istenilen bölümünü eksize ve kesi kapatılması, ameliyat için hayvan hazırlanması: prosedürü takip ayrıntıları. Apendiküler iskelet kemikleri ile karşılaştırıldığında, kaburga son derece erişilebilir. Komşu kirişleri doğal şekilde sabitlenmesini temin Ek olarak, herhangi bir dahili veya harici bir fiksatör gereklidir. cerrahi, piyasada mevcut kaynakları kullanır öğrenmek için basit ve hayvan tarafından iyi tolere edilir. işlem ile veya çevresindeki periost çıkarmadan gerçekleştirilir ve bu nedenle onarım Perioste- katkısı değerlendirilebilir. 2 ay - Sonuçlar periost kalınırsa, sağlam onarımı 1 oluştuğunu göstermektedir. Yapacaklarımız Bu protokol kullanımının bekliyoruzkaburga onarım içine araştırmaları teşvik ve bulgular vücudun etrafında diğer yerlerde kemik onarımı teşvik için yeni yollar geliştirilmesini kolaylaştıracak olacak.

Hayvanlar genellikle kesi iyi iyileşir, bu prosedürün hızla kurtarmak, ve hayvanlar normal davranış sergilerler. Prosedür piyasada mevcut kaynakları kullanır ve uygulama sonrasında, 1 saat altında tamamlanabilir. Deneysel verileri oluşturmak için bu prosedürü kullanmadan önce, herhangi bir büyük kemik parçaları rezeksiyon bölgesi içinde kalabilir olmadığını belirlemek için günlük 0 zaman noktasında rezeksiyon analiz etmek önemlidir. Bu genellikle özel radyasyon güvenlik onayları gerekti...

Watch this Scientific Journal Video about A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair at JoVE.com

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

The overall goal of this procedure is to successfully resect a portion of bone from the rib of a mouse. The procedure was developed as a model to study large-scale long bone repair.

Fare kaburga Rezeksiyonu için Cerrahi Prosedür: Büyük Ölçekli Uzun Kemik Onarım İçin Bir Model

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation ...

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Universidad de Cartagena UDEC

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13.0K Görüntüleme.  Keck School of Medicine, University of Southern California. The overall goal of this procedure is to successfully resect a portion of bone from the rib of a mouse. The procedure was developed as a model to study large-scale long bone repair.

Video: A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

A Mouse Distraction Osteogenesis Model

Distraction osteogenesis (DO) is a surgical procedure that involves skeletal tissue regeneration without cell transplantation. A DO model consists of the following three phases: the latency phase after osteotomy and placement of the external distractor; the distraction phase, wherein the separated bone ends are gradually and continuously distracted; and the consolidation phase. This custom-made distractor used for DO is comprised of two incomplete acrylic resin rings and an expansion screw. The process was initiated by making a mold with silicone impression material and then creating the custom-made distractor. Dental resin was poured into the formwork made of silicone impression material, and it was allowed to polymerize to create the incomplete resin rings required for the custom-made distractor. These rings were fixed with an expansion screw using transparent resin. The custom-made distractor created via this approach was attached to the tibia of mice. The tibia was fixed to the device using one pair of 25 G needles proximally, one pair of 27 G needles distally, and acrylic resin. After a latency period of 5 days, distraction was initiated at a rate of 0.2 mm/12 h. The lengthening was continued for 8 days, resulting in a total gap of 3.2 mm. The mice were sacrificed 4 weeks after distraction. Bone formation in the distraction gap was confirmed using both radiography and histology.

We present a mouse tibial distraction osteogenesis model developed using a custom-made distractor. The use of a mouse as an analysis target is advantageous for advancing research.

Bir fare oyalama Osteogenesis modeli

Distraction osteogenesis (DO) is a surgical procedure that involves skeletal tissue regeneration without cell transplantation. A DO model consists of the ...

Gelişim Biyolojisi

Mouse Pneumonectomy Model of Compensatory Lung Growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

Telafi Akciğer Büyüme Fare Pnömonektomi Modeli

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

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 ...

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 ...

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 ...

Sütür Genişletme Farelerinde Çekme Yükü Altında Kemik Yeniden Modellenmesinin 3D Görselleştirilmesi

A 3-D Visualization Technique for Bone Remodeling in a Suture Expansion Mouse Model

Craniofacial sutures play a crucial role beyond being fibrous joints connecting craniofacial bones; they also serve as the primary niche for calvarial and facial bone growth, housing mesenchymal stem cells and osteoprogenitors. As most craniofacial bones develop through intramembranous ossification, the sutures' marginal regions act as initiation points. Due to this significance, these sutures have become intriguing targets in orthopedic therapies like spring-assisted cranial vault expansion, rapid maxillary expansion, and maxillary protraction. Under orthopedic tracing force, suture stem cells are rapidly activated, becoming a dynamic source for bone remodeling during expansion. Despite their importance, the physiological changes during bone remodeling periods remain poorly understood. Traditional sectioning methods, primarily in the sagittal direction, do not capture the comprehensive changes occurring throughout the entire suture. This study established a standard mouse model for sagittal suture expansion. To fully visualize bone remodeling changes post-suture expansion, the PEGASOS tissue clearing method was combined with whole-mount EdU staining and calcium chelating double labeling. This allowed the visualization of highly proliferating cells and new bone formation across the entire calvarial bones following expansion. This protocol offers a standardized suture expansion mouse model and a 3-D visualization method, shedding light on the mechanobiological changes in sutures and bone remodeling under tensile force loading.

Yazar Spotlight: Kemik Yeniden Şekillenmesini Görselleştirmek için PEGASOS Doku Temizleme Tekniği 

This protocol presents a standardized suture expansion mouse model and a 3-D visualization method to study the mechanobiological changes of the suture and bone remodeling under tensile force loading.

Sütür Genişletme Fare Modelinde Kemiğin Yeniden Modellenmesi için 3 Boyutlu Görselleştirme Tekniği

Craniofacial sutures play a crucial role beyond being fibrous joints connecting craniofacial bones; they also serve as the primary niche for calvarial and ...