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

Nous remercions Sevil Essig, Stefanie Schroth, Verena Fischer, Katja Prystaz, Yvonne Hägele et Anne Subgang excellent soutien technique. Nous remercions également la Fondation de recherche allemande (CRC1149, INST40/499-1) et l’Allemagne AO Trauma Foundation pour le financement de cette étude.

animal toutes les expériences conformé au règlement international pour le soin et l’utilisation des animaux de laboratoire et ont été approuvées par les autorités de régulation régionales (no 1250, Regierungspräsidium Tübingen, Allemagne). Toutes les souris ont été maintenus en groupes de deux à cinq animaux par cage sur une lumière de 14 h, 10 h circadien sombre avec l’eau et la nourriture fournis ad libitum.
 1. préparation du matériel chirurgical et de pré-trai...

<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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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>

Modifications et dépannage :
L’objectif principal de cette étude était de décrire un protocole pour l’utilisation d’un fixateur externe compatible MRI pour ostéotomie du fémur chez la souris avec la possibilité de surveiller le développement des tissus cals longitudinalement pendant le processus de guérison des fractures des os endochondral. La conception d’un dispositif de montage sur mesure pour l’insertion du fixateur externe assuré une position normali...

Guérison de la fracture secondaire est la forme la plus courante de la guérison osseuse. C’est un processus complexe imitant des aspects spécifiques du endochondrale ontogénique ossification1,2,3. L’hématome au début de la fracture se compose principalement de cellules immunitaires, de granulation et de tissu fibreux. Faible tension d’oxygène et des souches biomécaniques hautes entravent une différenciation ostéoblastique à l’écart de la fracture, mais promouvoir la différenciation des cellules progénitrices en chondrocytes4,5,6. Ces cellules commencent à proliférer à l’endroit de la blessure pour former une matrice cartilagineuse assurant une stabilité initiale de l’OS fracturé. Durant la maturation des cals, devenus hypertrophiques, les chondrocytes subissent l’apoptose, ou trans-se différencient en ostéoblastes. La néovascularisation dans la zone de transition du cartilage-aux-os fournit des niveaux élevés d’oxygène, permettant la formation de tissu osseux7. Après le pont osseux de l’écart de la fracture, stabilité biomécanique est augmentée et ostéoclastique remodelage du cal fracture externe se produit pour avoir des os physiologique de contour et la structure3. Par conséquent, les quantités de tissu fibreux, cartilagineux et osseux dans le CAL de fracture fournissent des informations importantes concernant l’OS, processus de guérison. Perturbés ou retardé la guérison devient visible par des altérations du développement des tissus cals tant chez les humains et les souris8,9,10,11. Disponible en vivo techniques pour surveiller longitudinalement cals développement des tissus en préclinique fracture guérison des études à l’aide de petits animaux comprennent la radiographie numérique et µCT imagerie12,13. Toutefois, ces deux techniques ne sont capables de distinguer les tissus minéralisés et non minéralisée. En revanche, l’IRM fournit des tissus mous excellent contraste et pourrait donc être en mesure d’identifier les tissus mous et le cartilage dans le CAL de fracture.
Travaux antérieurs a montré des résultats prometteurs pour post mortem MRI chez la souris avec les fractures articulaires des IRM14 et in vivo chez des souris pendant intramembranaires défect osseux-guérison15. Cependant, les deux études a également déclaré limité contraste tissus et la résolution spatial. Nous avons précédemment démontré la faisabilité de haute résolution en vivo IAM pour une évaluation longitudinale de formation de cals doux pendant endochondrale murine fracture guérison16. Nous rapportons ici le protocole afin d’utiliser un fixateur externe compatible IRM-compatible pour ostéotomie du fémur chez la souris, afin de surveiller le développement des tissus cals longitudinalement au cours de la fracture de l’os endochondral processus de guérison. La conception d’un dispositif de montage sur mesure pour l’insertion du fixateur externe assuré une position normalisée lors d’analyses répétées.

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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="xl65" height="21" style="height:15.75pt">BioSpec 117/16 magnetic resonance imaging system</td>
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			<td class="xl64" style="width:149pt" width="199">117/16</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<td class="xl64" style="width:149pt" width="199">BH 021R</td>
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			<td class="xl65" height="21" style="height:15.75pt">Carbon steel scalpel no. 11/15</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<td class="xl64" style="width:149pt" width="199">BD210R</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<td class="xl64" style="width:149pt" width="199">BD025R</td>
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			<td class="xl64" style="width:149pt" width="199">2513026</td>
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			<td class="xl64" style="width:149pt" width="199">4928211</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">IOW-3704</td>
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			<td class="xl64" style="width:149pt" width="199">2706</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<td class="xl64" style="width:149pt" width="199">BD343R</td>
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			<td class="xl64" style="width:149pt" width="199">RIS.611.300-10</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<td class="xl64" style="width:149pt" width="199">BL911R</td>
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			<td class="xl65">Aesculap, Tuttlingen, Germany</td>
			<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="xl65" height="21" style="height:15.75pt">Oxygen, medical grade</td>
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			<td class="xl64" style="width:149pt" width="199">-</td>
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			<td class="xl65" height="21" style="height:15.75pt">Resolon 5/0</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

Guérison des fractures d’os endochondral est un processus complexe impliquant le développement de tissu fibreux, cartilagineux et osseux dans le CAL de fracture. La quantité des différents tissus dans le CAL fournit des informations importantes sur la rupture des progrès de la guérison. Disponible en vivo techniques longitudinalement surveille l’évolution de tissu de cals dans des études précliniques de guérison des fractures à l’aide de petits animaux comprennent la radiographie numérique et imagerie µCT. Toutefois, ces deux techniques ne sont en mesure de distinguer les tissus minéralisés et non minéralisée. Par conséquent, il est impossible de distinguer le cartilage de tissu fibreux. En revanche, l’imagerie par résonance magnétique (IRM) visualise les structures anatomiques basés sur leur teneur en eau et pourrait donc être en mesure d’identifier une façon non envahissante des tissus mous et le cartilage dans le CAL de fracture. Nous rapportons ici, l’utilisation d’un fixateur externe compatible IRM-compatible pour le fémur de la souris pour permettre des IRM au cours de la régénération osseuse chez la souris. Les expériences ont démontré que le fixateur et un dispositif de montage sur mesure permettent répétitives IRM, permettant ainsi une analyse longitudinale du développement des tissus fracture-Cal.

Tout d’abord, le succès de l’intervention chirurgicale peut être confirmé par l’analyse de l’IRM (voir exemple à la Figure 2). Tous les quatre épingles doivent être placés au milieu de la tige fémorale. La taille de l’écart de l’ostéotomie doit être comprise entre 0,3 et 0,5 mm. Si la taille de l’écart de l’ostéotomie varie grandement de ces valeurs, la souris doivent être exclue de l’analyse.
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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

In Vivo Évaluation du développement de cals de Fracture au cours de la guérison osseuse chez la souris en utilisant un dispositif d’ostéosynthèse IRM-compatible pour le fémur de souris

L’évaluation du développement des tissus dans le CAL de fracture au cours de la guérison osseuse endochondrale est indispensable pour surveiller le processus de guérison. Nous rapportons ici l’utilisation d’une résonance magnétique imagerie (IRM)-fixateur externe compatible pour le fémur de la souris pour permettre à MRI scans pendant la régénération osseuse chez la souris.

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

Évaluation longitudinale des souris Limb perte osseuse Hind après un traumatisme médullaire utilisant Novel, In vivo Méthodologie

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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Médecine

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

Développement d&#39;un environnement Audio-based jeux virtuels pour aider avec des compétences de navigation dans les aveugles

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

Séquentiel<em&gt; In vivo</em&gt; Imagerie des cellules souches / progénitrices ostéogéniques lors de la réparation de la fracture

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

Une méthode améliorée pour Rapid intubation de la trachée chez la souris

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

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Appauvrissement des cellules de souris de xénogreffes de tumeurs humaines améliore de manière significative l&#39;analyse en aval des cellules cibles

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

Un clou intramédullaire verrouillage pour Standardized Fixation de Fémur Ostéotomies pour l&#39;analyse normale et défectueuse des os de guérison chez la souris

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 ...

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

Une méthode d&#39;anesthésie, de chirurgie et de récolte pour l&#39;évaluation des vis transpediculaires en utilisant un<em&gt; In vivo</em&gt; Modèle de la colonne lombaire porcine

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

Un modèle mini-Invasive pour analyser Endochondral Fracture de guérison chez des souris dans des Conditions biomécaniques

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

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La génération des Fractures fémorales fermées chez la souris : un modèle pour étudier la guérison osseuse

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