Name: a minimally invasive model to analyze endochondral fracture healing in mice under standardized biomechanical conditions
Uploaded: 2018-03-22T15:00:00
Description: Watch this Scientific Journal Video about A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions at JoVE.com

Dette arbejde blev støttet af RISystem AG, Davos, Schweiz.

Alle procedurer blev udført Ifølge National Institutes of Health retningslinjer for brugen af forsøgsdyr og fulgt institutionelle retningslinjer (Landesamt für Verbraucherschutz, Zentralstelle Amtstierärztlicher Dienst, Saarbrücken, Tyskland).
1. forberedelse af kirurgiske instrumenter og implantater
<ol><li>Vælg en skalpel klinge (størrelse 15), en lille vatpind, fine pincet, en 27 G nål, en ikke-resorberbare 5-0 sutur, saks og en nål holder fra boksen microsurgical instrument.</l...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+size+and+stability+of+the+osteotomy+gap+on+the+success+of+fracture+healing.">Influence of size and stability of the osteotomy gap on the success of fracture healing.</a> J Orthop Res. 15 (4), 577-584 (1997).</li><li>Histing, T., 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=Characterization+of+the+healing+process+in+non-stabilized+and+stabilized+femur+fractures+in+mice.">Characterization of the healing process in non-stabilized and stabilized femur fractures in mice.</a> Arch Orthop Trauma Surg. 136 (2), 203-211 (2016).</li><li>Thompson, Z., Miclau, T., Hu, D., Helms, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+for+intramembranous+ossification+during+fracture+healing.">A model for intramembranous ossification during fracture healing.</a> J Orthop Res. 20 (5), 1091-1098 (2002).</li><li>Cheung, K. M., Kaluarachi, K., Andrew, G., Lu, W., Chan, D., Cheah, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+externally+fixed+femoral+fracture+model+for+mice.">An externally fixed femoral fracture model for mice.</a> J Orthop Res. 21 (4), 685-690 (2003).</li><li>Garcia, 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=A+new+technique+for+internal+fixation+of+femoral+fractures+in+mice:+impact+of+stability+on+fracture+healing.">A new technique for internal fixation of femoral fractures in mice: impact of stability on fracture healing.</a> J Biomech. 41 (8), 1689-1696 (2008).</li><li>Histing, T., 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=An+internal+locking+plate+to+study+intramembranous+bone+healing+in+a+mouse+femur+fracture+model.">An internal locking plate to study intramembranous bone healing in a mouse femur fracture model.</a> J Orthop Res. 28 (3), 397-402 (2010).</li><li>Garcia, 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=The+LockingMouseNail-a+new+implant+for+standardized+stable+osteosynthesis+in+mice.">The LockingMouseNail-a new implant for standardized stable osteosynthesis in mice.</a> J Surg Res. 169 (2), 220-226 (2011).</li><li>Histing, T., Klein, M., Stieger, A., Stenger, D., Steck, R., Matthys, R., Holstein, J. H., Garcia, P., Pohlemann, T., Menger, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+to+analyze+metaphyseal+bone+healing+in+mice.">A new model to analyze metaphyseal bone healing in mice.</a> J Surg Res. 178 (2), 715-721 (2012).</li><li>Histing, T., Menger, M. D., Pohlemann, T., Matthys, R., Fritz, T., Garcia, P., Klein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Intramedullary+Locking+Nail+for+Standardized+Fixation+of+Femur+Osteotomies+to+Analyze+Normal+and+Defective+Bone+Healing+in+Mice.">An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice.</a> J Vis Exp. (117), (2016).</li><li>Hiltunen, A., Vuorio, E., Aro, H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+standardized+experimental+fracture+in+the+mouse+tibia.">A standardized experimental fracture in the mouse tibia.</a> J Orthop Res. 11 (2), 305-312 (1993).</li><li>Manigrasso, M. B., O'Connor, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+closed+femur+fracture+model+in+mice.">Characterization of a closed femur fracture model in mice.</a> J Orthop Trauma. 18 (10), 687-695 (2004).</li><li>Holstein, J. H., Menger, M. D., Culemann, U., Meier, C., Pohlemann, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+locking+femur+nail+for+mice.">Development of a locking femur nail for mice.</a> J Biomech. 40 (1), 215-219 (2007).</li><li>Claes, L. E., 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=Effects+of+mechanical+factors+on+the+fracture+healing+process.">Effects of mechanical factors on the fracture healing process.</a> Clin Orthop Relat Res. 355 Suppl, S132-S147 (1998).</li></ol>

Kritiske trin i den kirurgiske procedure er at finde den korrekte indgang til skrue implantation i midten af femur condyles på den intercondylar notch samt den optimale orientering af nålen parallelt med knogle akse for rivning af den intramedullære hulrum. For at undgå en forkert post holdning, skal kirurgen forberede hak indtil en optimal udsigt er opnået. For at styre retning under rivning, bør lårben musenes holdes med fingrene i en stabil stilling. Et yderligere vigtigt skridt er indsættelsen af skruen i lå...

Knogle helbredende studier i mus er i stor efterspørgsel på grund af et bredt spektrum af antistoffer og genetisk modificerede dyr. Disse fakta gør det muligt for at studere de molekylære mekanismer af knogle heling1. I de seneste år udviklet forskellige knogle heling modeller for mus har været2. Disse modeller kan være opdelt i åbne modeller, hvor knoglen er osteotomized ved hjælp af en åben laterale kirurgisk tilgang og i lukkede modeller, hvor knoglen er brækket baseret på fraktur model indført af Bonnares og Einhorn3. Brug af denne teknik, en standardiseret tværgående fraktur kan fremstilles af en 3-punkts bøjning enhed og intramedullært brystimplantater kan indsættes gennem et lille mediale parapatellar snit i en minimalt invasiv teknik at undgå et stort blødt væv traumer.
Intramedullære skruen kan anvendes til lukket fraktur stabilisering i mus. Skruen tilbyder roterende og aksial stabilitet. Dette opnås ved fraktur kompression gennem en proksimal tråd og en distal hoved4. Yderligere fordele ved skruen er den simple kirurgiske teknik, den lave kvalitet af invasivity, lave implantat vægt og, især, en højere stabilitet giver standardiserede og kontrolleret biomekaniske forhold i forhold til andre intramedullært implantater5. I virkeligheden, i de mest lukkede fraktur modeller, fragmenter er stabiliseret kun af simple pins, som er forbundet med en komplet mangel på roterende og aksial stabilitet og en høj risiko for pin og også fraktur dislokation. Dette kan markant indflydelse på helingsprocessen, som kan medføre forsinket heling eller ikke-union dannelsen.
Det er velkendt, at stabilitet af frakturfiksation har en enorm indflydelse på den helbredende proces6,7. En høj rigid fiksation resulterer i intramembranous healing, mens en mindre stive fiksation, som kan give micromovements i fraktur gap, resulterer i endochondral healing. Stabilisering af fraktur med intramedullært skruen viser overvejende en endochondral healing med en stor mængde af hård hud væv, især efter 2 ugers frakturheling. Mulighed for at høste en stor mængde af hård hud væv muliggør analysen af flere parametre ved hjælp af forskellige teknikker.
Her, rapport vi om udformningen og anvendelsen af intramedullært skruen i mus, og om fordele og ulemper i eksperimentelle undersøgelser normale endochondral knogleheling.

<table>
	<tbody>
		<tr>
			<td>Mouse Screw</td>
			<td>RISystem AG</td>
			<td>221,100</td>
			<td></td>
		</tr>
		<tr>
			<td>Guide wire</td>
			<td>RISystem AG</td>
			<td>521,100</td>
			<td></td>
		</tr>
		<tr>
			<td>Centering bit</td>
			<td>RISystem AG</td>
			<td>590,205</td>
			<td></td>
		</tr>
		<tr>
			<td>Hand drill</td>
			<td>RISystem AG</td>
			<td>390,130</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton-Swab (150 mm, small head)</td>
			<td>Fink Walter GmbH</td>
			<td>8822428</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture (5-0 Prolene)</td>
			<td>Ethicon</td>
			<td>8614H</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Braun Aesculap AG &#38;CoKG</td>
			<td>BD520R</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Braun Aesculap AG &#38;CoKG</td>
			<td>BC100R</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Braun Aesculap AG &#38;CoKG</td>
			<td>BM024R</td>
			<td></td>
		</tr>
		<tr>
			<td>27 G needle</td>
			<td>Braun Melsungen AG</td>
			<td>9186182</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel blade size 15</td>
			<td>Braun Aesculap AG &#38;CoKG</td>
			<td>16600525</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat radiator</td>
			<td>Sanitas</td>
			<td>605.25</td>
			<td></td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Asid bonz GmbH</td>
			<td>NDXZ10</td>
			<td></td>
		</tr>
		<tr>
			<td>Eye lubricant</td>
			<td>Bayer Vital GmbH</td>
			<td>2182442</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Bayer Vital GmbH</td>
			<td>1320422</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Serumwerke Bernburg</td>
			<td>7005294</td>
			<td></td>
		</tr>
		<tr>
			<td>Tramadol</td>
			<td>Gr&#252;nenthal GmbH</td>
			<td>2256241</td>
			<td></td>
		</tr>
		<tr>
			<td>Disinfection solution (SoftaseptN)</td>
			<td>Braun Melsungen AG</td>
			<td>8505018</td>
			<td></td>
		</tr>
		<tr>
			<td>CD-1 mice</td>
			<td>Charles River</td>
			<td>22</td>
			<td></td>
		</tr>
		<tr>
			<td>X-ray Device</td>
			<td>Faxitron MX-20, Faxitron X-ray Corporation</td>
			<td>2321A0988</td>
			<td></td>
		</tr>
		<tr>
			<td>Fracture device small</td>
			<td>RISystem AG</td>
			<td>891,100</td>
			<td></td>
		</tr>
	</tbody>
</table>

a minimally invasive model to analyze endochondral fracture healing in mice under standardized biomechanical conditions

Knogle helbredende modeller er nødvendige for at analysere de komplekse mekanismer af fraktur healing for at forbedre den kliniske fracture behandling. I det sidste tiår, blev øget brug af musemodeller i ortopædiske forskning bemærket, sandsynligvis fordi musemodeller tilbyder et stort antal genetisk modificerede stammer og specielle antistoffer til analyse af molekylære mekanismer af frakturheling. Du kan styre de biomekaniske forhold ved er godt karakteriseret osteosyntese teknikker obligatorisk, også i mus. Her, rapport vi om design og brug af en lukket knogle heling model til at stabilisere femur frakturer i mus. Intramedullære skruen, i medicinsk-grade rustfrit stål, giver gennem fraktur komprimering en aksial og roterende stabilitet i forhold til det meste brugte simple intramedullært pins, som viser en fuldstændig mangel på aksial og roterende stabilitet. Stabiliteten opnås ved intramedullært skruen giver mulighed for analyse af endochondral healing. En stor mængde af hård hud væv, modtaget efter stabilisering med skruen, tilbyder ideelle betingelser for at høste væv til biokemiske og molekylære analyser. En yderligere fordel ved brugen af skruen er det faktum, at skruen kan indsættes i lårbenet med en minimalt invasiv teknik uden inducerende skade på det bløde væv. Afslutningsvis, er at skruen en unik implantat, der kan ideelt bruges i lukket fraktur healing modeller tilbyder standardiseret biomekaniske forhold.

Driftstid fra hud indsnit til sår lukning var 20 min. Kirurgi kan udføres uden et stereo-mikroskop. Efter operationen, var at dyrene overvåges dagligt. Postoperativ analgesi blev afbrudt efter 2 dage, fordi ingen af dyrene viste tegn på smerter efter denne periode. Dyrene viste også normale vægtbærende senest 2 dage efter operationen. Sårinfektioner blev ikke observeret i hele observationsperioden.
Radiologiske analyser ...

Watch this Scientific Journal Video about A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions at JoVE.com

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

Denne protokol beskriver en minimalt invasiv osteosyntese teknik bruger et intramedullære skrue for standardiseret stabilisering af femur frakturer, som kan bruges til at analysere endochondral knogle heling i mus.

En minimalt invasiv Model til at analysere Endochondral fraktur Healing i mus Under standardiserede biomekaniske forhold

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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