Name: in vivo evaluation of fracture callus development during bone healing in mice using an mri-compatible osteosynthesis device for the mouse femur
Uploaded: 2017-11-14T15:00:00
Description: Watch this Scientific Journal Video about In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur at JoVE.com

Agradecemos Sevil Essig, Stefanie Schroth, Verena Fischer, Katja Prystaz, Yvonne Hägele e Anne Subgang excelente suporte técnico. Agradecemos também a Fundação de pesquisa alemã (CRC1149, INST40/499-1) e a Alemanha AO Trauma Foundation para financiamento deste estudo.

animal todos os experimentos cumprimento de regulamentos internacionais para o cuidado e o uso de animais de laboratório e foram aprovados pelas autoridades reguladoras regionais (n. º 1250, Regierungspräsidium Tübingen, Alemanha). Todos os ratos foram mantidos em grupos de dois a cinco animais por gaiola a luz 14-h, 10 h a escuro ritmo circadiano com água e alimentos fornecidos ad libitum.
 1. preparação do Material cirúrgico e pré-tratamento dos ratos <ol><li>esterilize ...

<ol>
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P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cartilage+to+bone+transformation+during+fracture+healing+is+coordinated+by+the+invading+vasculature+and+induction+of+the+core+pluripotency+genes.">Cartilage to bone transformation during fracture healing is coordinated by the invading vasculature and induction of the core pluripotency genes.</a> Development. 144 (2), 221-234 (2017).</li><li>Hankenson, K. D., Zimmerman, G., Marcucio, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+perspectives+of+delayed+fracture+healing.">Biological perspectives of delayed fracture healing.</a> Injury. 45, 8-15 (2014).</li><li>Meyer, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age+and+ovariectomy+impair+both+the+normalization+of+mechanical+properties+and+the+accretion+of+mineral+by+the+fracture+callus+in+rats.">Age and ovariectomy impair both the normalization of mechanical properties and the accretion of mineral by the fracture callus in rats.</a> J Orthop Res. 19 (3), 428-435 (2001).</li><li>Nikolaou, V. S., Efstathopoulos, N., Kontakis, G., Kanakaris, N. K., Giannoudis, P. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+osteoporosis+in+femoral+fracture+healing+time.">The influence of osteoporosis in femoral fracture healing time.</a> Injury. 40 (6), 663-668 (2009).</li><li>Haffner-Luntzer, M., Kovtun, A., Rapp, A. 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A., Bertone, A. L., Wassenaar, P. A., Weisbrode, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+for+the+study+of+articular+fracture+healing.">Rodent models for the study of articular fracture healing.</a> J Invest Surg. 20 (2), 87-95 (2007).</li><li>Taha, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+the+efficacy+of+MRI+for+detection+of+changes+in+bone+morphology+in+a+mouse+model+of+bone+injury.">Assessment of the efficacy of MRI for detection of changes in bone morphology in a mouse model of bone injury.</a> J Magn Reson Imaging. 38 (1), 231-237 (2013).</li><li>Haffner-Luntzer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+high-resolution+In+Vivo+MRI+for+longitudinal+analysis+of+endochondral+fracture+healing+in+mice.">Evaluation of high-resolution In Vivo MRI for longitudinal analysis of endochondral fracture healing in mice.</a> PLoS One. 12 (3), 0174283 (2017).</li><li>Beckmann, N., Falk, R., Zurbrugg, S., Dawson, J., Engelhardt, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Macrophage+infiltration+into+the+rat+knee+detected+by+MRI+in+a+model+of+antigen-induced+arthritis.">Macrophage infiltration into the rat knee detected by MRI in a model of antigen-induced arthritis.</a> Magn Reson Med. 49 (6), 1047-1055 (2003).</li><li>Al Faraj, ., Shaik A, S. u. l. t. a. n. a., Pureza, A., A, M., Alnafea, M., Halwani, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preferential+macrophage+recruitment+and+polarization+in+LPS-induced+animal+model+for+COPD:+noninvasive+tracking+using+MRI.">Preferential macrophage recruitment and polarization in LPS-induced animal model for COPD: noninvasive tracking using MRI.</a> PLoS One. 9 (3), 90829 (2014).</li><li>Rolle, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ImmunoPET/MR+imaging+allows+specific+detection+of+Aspergillus+fumigatus+lung+infection+in+vivo.">ImmunoPET/MR imaging allows specific detection of Aspergillus fumigatus lung infection in vivo.</a> Proc Natl Acad Sci U S A. 113 (8), 1026-1033 (2016).</li><li>Niemeyer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+tracking+of+human+haemopoietic+CD34(+)+stem+cells+in+vivo+in+immunodeficient+mice+by+using+magnetic+resonance+imaging.">Non-invasive tracking of human haemopoietic CD34(+) stem cells in vivo in immunodeficient mice by using magnetic resonance imaging.</a> Eur Radiol. 20 (9), 2184-2193 (2010).</li></ol>

Modificações e solução de problemas:
O principal objetivo deste estudo foi descrever um protocolo para o uso de fixador externo compatível com o MRI para osteotomia do fêmur no mouse com a capacidade de monitorar o desenvolvimento de tecido calo longitudinalmente durante o processo de consolidação da fratura endocondral. O projeto de um dispositivo de montagem sob medida para a inserção do fixador externo assegurado uma posição padronizada durante as varreduras rep...

Consolidação da fratura secundária é a forma mais comum de consolidação óssea. É um processo complexo, imitando a aspectos específicos da32,1,do ossificação endocondral ontogenic. O hematoma de fratura precoce consiste predominantemente de células do sistema imunológico, granulação e tecido fibroso. Tensão de oxigênio de baixa e altas tensões biomecânicas dificultam a diferenciação osteoblástica na gap da fratura, mas promovem a diferenciação de células progenitoras em condrócitos4,5,6. Essas células começam a proliferar no local da lesão para formar uma matriz cartilaginosa, fornecendo estabilidade inicial do osso fraturado. Durante a maturação do calo, condrócitos se tornar hipertróficas, passam por apoptose, ou trans-se diferenciar em osteoblastos. Neovascularização na zona de transição de cartilagem-de-osso fornece níveis elevados de oxigênio, permitindo a formação de tecido ósseo,7. Depois da ponte óssea do fosso de fratura, estabilidade biomecânica é aumentada e osteoclástica remodelação do calo externo fratura ocorre para ganhar óssea fisiológica contorno e estrutura3. Portanto, a quantidade de tecido fibroso, cartilaginoso e ósseo no calo de fratura fornece informações importantes sobre a processo de consolidação. Perturbado ou retardado cura torna-se visível por alterações do desenvolvimento do tecido de calo tanto em humanos e ratos8,9,10,11. Disponível na vivo técnicas para monitorar longitudinalmente o desenvolvimento de tecido calo ósseo em fratura pré-clínicos cura estudos utilizando pequenos animais incluem radiografia digital e µCT de imagem12,13. No entanto, ambas as técnicas só são capazes de discriminar entre o tecido mineralizado e não-mineralizado. Em contraste, ressonância fornece contraste excelente de tecidos moles e, portanto, pode ser capaz de identificar os tecidos moles e cartilagem no calo de fratura.
Trabalhos anteriores mostraram resultados promissores para post mortem MRI em camundongos com fraturas articulares14 e na vivo MRI em ratos durante intramembranosa defeito ósseo-cura15. No entanto, ambos os estudos também declarou limitada contraste espacial, resolução e tecido. Nós anteriormente demonstrou a viabilidade da RM de alta resolução na vivo para avaliação longitudinal da formação de calo macio durante fratura endocondral murino16de cura. Aqui, nós relatamos o protocolo para o uso de fixador externo compatível com o MRI para osteotomia de fêmur em ratos a fim de monitorar o desenvolvimento de tecido calo longitudinalmente durante a fratura endocondral, processo de cura. O projeto de um dispositivo de montagem sob medida para a inserção do fixador externo assegurado uma posição padronizada durante as varreduras repetidas.

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			<td class="xl65" height="21" style="width:254pt;height:15.75pt" width="338">Anaesthesia tube</td>
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			<td class="xl64" style="width:149pt" width="199">117/16</td>
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			<td class="xl64" style="width:149pt" width="199">BH 021R</td>
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			<td class="xl64" style="width:149pt" width="199">2513026</td>
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			<td class="xl65">RISystem, Davos, Switzerland</td>
			<td class="xl64" style="width:149pt" width="199">RIS.590.110.25</td>
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			<td class="xl64" style="width:149pt" width="199">RIS.390.130-01</td>
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			<td class="xl64" style="width:149pt" width="199">2706</td>
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			<td class="xl64" style="width:149pt" width="199">BB078R</td>
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			<td class="xl65" height="21" style="height:15.75pt">Octenisept</td>
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			<td class="xl64" style="width:149pt" width="199">-</td>
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			<td class="xl64" style="width:149pt" width="199">-</td>
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in vivo evaluation of fracture callus development during bone healing in mice using an mri-compatible osteosynthesis device for the mouse femur

Endocondral fratura cura é um processo complexo que envolve o desenvolvimento de tecido fibroso, cartilaginoso e ósseo no calo de fratura. A quantidade de tecidos diferentes em calo ósseo fornece informações importantes sobre a fratura cura progresso. Disponível na vivo técnicas para monitorar longitudinalmente o desenvolvimento de tecido de calo em estudos pré-clínicos consolidação da fratura, utilizando pequenos animais incluem radiografia digital e imagem latente de µCT. No entanto, ambas as técnicas só são capazes de distinguir entre o tecido mineralizado e não-mineralizado. Por conseguinte, é impossível discriminar cartilagem de tecido fibroso. Em contraste, ressonância magnética (MRI) visualiza estruturas anatômicas com base no seu teor de água e, portanto, pode ser capaz de identificar canaliza o tecido mole e cartilagem no calo de fratura. Aqui, nós relatamos o uso do fixador externo compatível com o MRI para o fêmur do mouse para permitir ressonâncias durante a regeneração óssea em ratos. Os experimentos demonstraram que o fixador externo e um dispositivo de fixação feito por permitam ressonâncias repetitivas, possibilitando assim a análise longitudinal do desenvolvimento do tecido de calo de fratura.

Em primeiro lugar, o sucesso do procedimento cirúrgico pode ser confirmado pela análise das varreduras de MRI (ver exemplo na Figura 2). Todos os quatro pinos deverá situar-se no meio da diáfise femoral. O tamanho da lacuna a osteotomia deve ser entre 0.3-0.5 mm. Se o tamanho da lacuna osteotomia varia muito desses valores, o mouse deve ser excluído uma análise mais aprofundada.
Em segundo lug...

Watch this Scientific Journal Video about In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur at JoVE.com

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

Na Vivo Avaliação do desenvolvimento de calo de fratura durante a consolidação óssea em ratos usando um dispositivo de osteossíntese compatível com MRI para o fêmur de Mouse

A avaliação do desenvolvimento do tecido no calo de fratura durante a cicatrização de osso endocondral é essencial para monitorar o processo de cicatrização. Aqui, nós relatamos o uso de uma ressonância magnética (MRI)-fixador externo compatível para o fêmur do mouse para permitir MRI varreduras durante a regeneração óssea em ratos.

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

Research

JoVE Journal

Medicine

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

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

Avaliação longitudinal do rato Perda Hind Limb óssea após lesão medular usando Novel, In vivo Metodologia,

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher ...

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Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

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Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Sequential In vivo Imaging of Osteogenic Stem/Progenitor Cells During Fracture Repair

Sequential In vivo Imaging of Osteogenic Stem/Progenitor Cells During Fracture Repair

Seqüente<em&gt; In vivo</em&gt; Imagem de osteogênicas células-tronco / progenitoras durante o reparo de fratura

Bone turns over continuously and is highly regenerative following injury. Osteogenic stem/progenitor cells have long been hypothesized to exist, but in ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Um método melhorado para rápida de intubação da traqueia em Ratos

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

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The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

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

Uma haste intramedular de bloqueio para Padronizado Fixação do fêmur Osteotomias para analisar normal e defeituosa cicatrização óssea em ratos

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

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

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

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

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Método de anestesia, cirurgia e colheita para avaliação de parafusos transpediculares usando um In Vivo Modelo de espinha lombar porcina

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

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

Um modelo minimamente invasivo para analisar a fratura endocondral cura em ratos sob condições biomecânicas padronizadas

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

A geração de fraturas fechadas de fêmur em ratos: um modelo para estudar a consolidação óssea

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