Name: an efficient and reproducible protocol for distraction osteogenesis in a rat model leading to a functional regenerated femur
Uploaded: 2017-10-23T15:00:00
Description: Watch this Scientific Journal Video about An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur at JoVE.com

这项工作得到了 CNRS Mecabio 挑战的支持和资助。
作者感谢动物护理技术员在整个过程中照顾动物。作者还通过 IVTV 承认里昂中心。感谢马乔里 Sweetko 的语言版本。
我们感激 Marylène Lallemand、Cécile Génovésio 和帕特里克. 劳伦对这项实验研究的贡献。

所描述的所有程序均由 Aix-马赛机构动物保育和使用委员会和法国研究部批准, 并在马赛医学院的常规动物院进行 (法国). 
 1. 根据以下准则定义外部固定器的功能规范 <ol> <li> 优化骨骼锚固. 
	<ol> <li> 植入 half-threaded 针脚, 其直径 (螺纹部分) 为 1 mm. </li>
	</ol> </li> <li> 选择设计以减少对动物的不适. 
	<ol> <li> 选择适合于 7723 mm 3 (32 mm x 19 mm x ...

<ol>
	<li>Jauregui, J. J., Ventimiglia, A. V., Grieco, P. W., Frumberg, D. B., Herznberg, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerate+Bone+stimulation+following+limb+lengthening:+a+meta-analysis.">Regenerate Bone stimulation following limb lengthening: a meta-analysis.</a> BMC Musculoskelet Disord. 17 (1), 407 (2016).</li><li>Morcos, M. W., Al-Jallad, H., Hamdy, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+review+of+Adipose+Stem+Cells+and+Their+Implication+in+Distraction+Osteogenesis+and+Bone+Regeneration.">Comprehensive review of Adipose Stem Cells and Their Implication in Distraction Osteogenesis and Bone Regeneration.</a> Biomed Res Int. 2015 (842975), 1-20 (2015).</li><li>Cans&#252;, E., &#220;nal, M. B., Parmaksizoglu, F., G&#252;rcan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distraction+lengthening+of+the+proximal+phalanx+in+distal+thumb+amputations.">Distraction lengthening of the proximal phalanx in distal thumb amputations.</a> Acta Orthop Traumatol Turc. 49 (3), 227-232 (2014).</li><li>Singh, M., Vashistha, A., Chaudhary, M., Kaur, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+Basis+of+Distraction+Osteogenesis+-+A+Review.">Biological Basis of Distraction Osteogenesis - A Review.</a> J Oral Maxillofac Surg Med Pathol. 28 (1), 1-7 (2016).</li><li>Sailhan, F., Gleyzolle, B., Parot, R., Guerini, H., Viguier, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rh-BMP-2+in+Distraction+Osteogenesis:+Dose+Effect+and+Premature+Consolidation.">Rh-BMP-2 in Distraction Osteogenesis: Dose Effect and Premature Consolidation.</a> Injury. 41 (7), 680-686 (2010).</li><li>Schiller, J. R., Moore, D. C., Ehrlich, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+Lengthening+Rate+Decreases+Expression+of+Fibroblast+Growth+Factor+2,+Platelet-Derived+Growth+Factor,+Vascular+Endothelial+Growth+Factor,+and+CD31+in+a+Rat+Model+of+Distraction+Osteogenesis.">Increased Lengthening Rate Decreases Expression of Fibroblast Growth Factor 2, Platelet-Derived Growth Factor, Vascular Endothelial Growth Factor, and CD31 in a Rat Model of Distraction Osteogenesis.</a> J Pediatr Orthop. 27 (8), 961-968 (2007).</li><li>Aronson, J., Shen, X. C., Skinner, R. A., Hogue, W. R., Badger, T. M., Lumpkin, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+Model+of+Distraction+Osteogenesis.">Rat Model of Distraction Osteogenesis.</a> J Orthop Res. 15 (2), 221-226 (1997).</li><li>Aida, T., Yoshioka, I., Tominaga, K., Fukuda, J. <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+latency+period+in+a+rabbit+mandibular+distraction+osteogenesis.">Effects of latency period in a rabbit mandibular distraction osteogenesis.</a> Int J Oral Maxillofac Surg. 32, 54-62 (2003).</li><li>Lammens, J., Liu, Z., Aerssend, J., Dequeker, J., Fabry, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=distraction+Bone+Healing+Versus+Osteotomy+Healing:+A+Comparative+Biochemical+Analysis.">distraction Bone Healing Versus Osteotomy Healing: A Comparative Biochemical Analysis.</a> J Bone Miner Res. 13 (2), 279-286 (1998).</li><li>Isefuku, S., Joyner, C. J., Simpson, A. H. R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Murine+Model+of+Distraction+Osteogenesis.">A Murine Model of Distraction Osteogenesis.</a> Bone. 27 (5), 661-665 (2000).</li><li>Eberson, C. P., Hogan, K. A., Moore, D. C., Ehrlich, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+Low-Intensity+Ultrasound+Stimulation+on+Consolidation+of+the+Regenerate+Zone+in+a+Rat+Model+of+Distraction+Osteogenesis.">Effect of Low-Intensity Ultrasound Stimulation on Consolidation of the Regenerate Zone in a Rat Model of Distraction Osteogenesis.</a> J Pediatr Orthop. 23 (1), 46-51 (2003).</li><li>&#214;zdel, A., Saris&#246;zen, B., Yal&#231;inkaya, U., Demirag, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+HIF+Stabilizer+on+Distraction+Osteogenesis.">The Effect of HIF Stabilizer on Distraction Osteogenesis.</a> Acta Orthop Traumatol Turc. 49 (1), 80-84 (2015).</li><li>Takamine, Y., 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=Distraction+Osteogenesis+Enhanced+by+Osteoblastlike+Cells+and+Collagen+Gel.">Distraction Osteogenesis Enhanced by Osteoblastlike Cells and Collagen Gel.</a> Clin Orthop Relat Res. 399, 240-246 (2002).</li><li>Wang, X., Zhu, S., Jiang, X., Li, Y., Song, D., Hu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systemic+Administration+of+Lithium+Improves+Distracted+Bone+Regeneration+in+Rats.">Systemic Administration of Lithium Improves Distracted Bone Regeneration in Rats.</a> Calcif Tissue Int. 96 (6), 534-540 (2015).</li><li>Xu, J., 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=Human+Fetal+Mesenchymal+Stem+Cell+Secretome+Enhances+Bone+Consolidation+in+Distraction+Osteogenesis.">Human Fetal Mesenchymal Stem Cell Secretome Enhances Bone Consolidation in Distraction Osteogenesis.</a> Stem Cell Res Ther. 7 (1), (2016).</li><li>Yasui, N., 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=Three+Modes+of+Ossification+during+Distraction+Osteogenesis+in+the+Rat.">Three Modes of Ossification during Distraction Osteogenesis in the Rat.</a> Bone Joint J. 79 (5), 824-830 (1997).</li><li>Chang, F., 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=Stimulation+of+EP4+Receptor+Enhanced+Bone+Consolidation+during+Distraction+Osteogenesis.">Stimulation of EP4 Receptor Enhanced Bone Consolidation during Distraction Osteogenesis.</a> J Orthop Res. 25 (2), 221-229 (2007).</li><li>Nyman, J. S., 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=Quantitative+Measures+of+Femoral+Fracture+Repair+in+Rats+Derived+by+Micro-Computed+Tomography.">Quantitative Measures of Femoral Fracture Repair in Rats Derived by Micro-Computed Tomography.</a> J Biomech. 42 (7), 891-897 (2009).</li><li>Hsu, J. T., Wang, S. P., Huang, H. L., Chen, Y. J., Wu, J., Tsai, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+assessment+of+trabecular+bone+parameters+and+cortical+bone+strength:+a+comparison+of+micro-CT+and+dental+cone-beam+CT.">The assessment of trabecular bone parameters and cortical bone strength: a comparison of micro-CT and dental cone-beam CT.</a> J Biomech. 46, 2611-2618 (2013).</li><li>Xue, J., 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=NELL1+Promotes+High-Quality+Bone+Regeneration+in+Rat+Femoral+Distraction+Osteogenesis+Model.">NELL1 Promotes High-Quality Bone Regeneration in Rat Femoral Distraction Osteogenesis Model.</a> Bone. 48 (3), 485-495 (2011).</li><li>Mark, H., Bergholm, J., Nilsson, A., Rydevik, B., Str&#246;mberg, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+External+Fixation+Method+and+Device+to+Study+Fracture+Healing+in+Rats.">An External Fixation Method and Device to Study Fracture Healing in Rats.</a> Acta Orthop. 74 (4), 476-482 (2003).</li></ol>

这项研究描述了一个可复制的协议, 包括一个适当的外固定器设计的大鼠肢体延长, 与详细的手术技术, 允许动物的生理负重后, 去除外固定架。我们的协议导致了一个功能性的再生骨骼。经过47天的巩固, 去除自制的外固定器和2天的生理负重, 动物并没有诱发任何骨折的再生愈伤组织。由于微 CT 重建, 完全架桥的证据证实了再生愈伤组织的功能。先前的研究清楚地证明了钙化桥接的存在与再生愈伤?...

牵引成骨术 (做) 是一种外科技术广泛使用临床1,2,3,4在人为更低的1,2和上部3肢体延长,治疗后骨不连, 或再生骨缺损后骨肿瘤切除术以及颌面部重建4。在骨内固定支架置入后, 可导致骨再生。osteotomized 四肢通过逐渐分心2 , 以达到预期的伸长距离。巩固期间跟随, 在期间没有更多伸长。
执行过程分为三不同的阶段: 延迟、分心和整合。通常, 7 天的潜伏期是在截骨4之后开始的。这允许骨骼修复开始愈合过程的初始步骤4。潜伏期后, 牵引力被应用到再生愈伤组织和周围软组织的一个分心期1,2,4。当达到预期的伸长率, 分心停止和巩固期开始。在此期间, 外固定器保持, 直到再生骨功能足以支持其去除。
不同的参数对骨修复有影响, 如长度和伸长率, 外固定器的类型, 分心的频率, 巩固期的长短, 或机械应力类型应用于分散的愈伤组织。举例来说, 延长的速度和频率可能导致过早的合并5或通过创建不可恢复的损伤, 如坏死组织或囊肿在胼胝体内,6,7。
许多协议已经应用于不同的动物模型8,9,10 , 以研究骨骼修复过程并最大限度地巩固骨骼。在大鼠中, 大多数研究11、12、13、14、15 , 重点讨论如何通过加速愈伤组织合并来缩短执行协议。其中一些实验研究使用了外部固定架已经商业化可用于人类临床应用的5,13,15,16。然而, 这些类型的外固定器不适合做的大鼠股骨, 这表现出不同的解剖特征, 从人的股骨。此外, 只有少数研究清楚地证明了它们的协议在获得功能性再生骨716中的效率。因此, 很难比较不同的研究结果, 由于他们的不同协议和缺乏有关外部固定器的信息12,13,14,17。
因此, 这项研究的目的是描述, 在大鼠模型, 一个有效的和可重复的协议做股骨, 导致功能性再生骨。为此, 我们设计了一个自制和易于使用的外固定器, 特别是对大鼠股骨, 这是我们在这个协议中详细描述。在起草该装置的技术规格时, 我们考虑到了机械应力分布良好和避免残余应力产生的所有基本约束。技术规格包括一个适当的几何为设备允许纯净的牵引力量在骨头和周围的组织, 适当的重量为动物的步态, 控制骨头伸长的长度和好对骨头段的对准在销钉和骨骼的交叉处不产生剪切应力。此外, 该装置必须在不受干扰、生物相容性和消毒无损伤的情况下使用。经过7周的整合后, 本议定书对大鼠股骨做了一个功能性再生骨, 证明了动物的生理负重没有骨折的再生愈伤组织后外固定架。动物的生理步态与再生愈伤组织和 x 射线分析的显微 CT 分析得到的建筑参数一致。

<table>
	<tbody>
		<tr>
			<td>K&#233;tamine</td>
			<td>Renaudin</td>
			<td>578 540-2</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>M&#233;d&#233;tomidine</td>
			<td>Virbac</td>
			<td>6799091</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Sevoflurane</td>
			<td>Centravet</td>
			<td>567 477-2</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Indivor France</td>
			<td>3400932731060</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Enrofloxacine</td>
			<td>ChannelPharmaceutical Facturing</td>
			<td>FR/V/4955220</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Piezotome</td>
			<td>Satelec Acteon</td>
			<td>F57510</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pet pad</td>
			<td>Therasage</td>
			<td>AL8365936</td>
			<td>Supply by the animal house</td>
		</tr>
		<tr>
			<td>Dental X-ray</td>
			<td>S.A.R.L Innovation m&#233;dicales et dentaires</td>
			<td>WYZ - BLUEX</td>
			<td></td>
		</tr>
		<tr>
			<td>Winiwix Software</td>
			<td>Softys Dental</td>
			<td>PFT</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-CT system</td>
			<td>nanoScan SPECT/CT</td>
			<td>GEIT-31105EN (05/14)</td>
			<td>Subcontract by IVTV central Lyon</td>
		</tr>
		<tr>
			<td>Micro-CT analysis Software</td>
			<td>phoenix datos X2 reconstruction</td>
			<td>none</td>
			<td>Free software</td>
		</tr>
		<tr>
			<td>Electric razor</td>
			<td>Brawn</td>
			<td>GT415</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Senn&#8217;s retractors</td>
			<td>Word Precision Instruments</td>
			<td>501718</td>
			<td>Blunt version</td>
		</tr>
		<tr>
			<td>Betadine Solution</td>
			<td>Mundipharma Medical Company</td>
			<td>D08AG02</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Resorbable suture thread (5.0)</td>
			<td>Ethicon</td>
			<td>JV1023</td>
			<td>Supply by animal house</td>
		</tr>
		<tr>
			<td>Rugine</td>
			<td>Word Precision Instruments</td>
			<td>503406</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayo-Hegar needle holder</td>
			<td>Word Precision Instruments</td>
			<td>V503382</td>
			<td></td>
		</tr>
		<tr>
			<td>Metal drill</td>
			<td>Beuterlock</td>
			<td>V020944018003</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Olsen-Hegar Needle-holder</td>
			<td>Word Precision Instruments</td>
			<td>501989</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayo scissor</td>
			<td>Word Precision Instruments</td>
			<td>501752</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Word Precision Instruments</td>
			<td>500236</td>
			<td></td>
		</tr>
		<tr>
			<td>Sprague-Dawley</td>
			<td>Janvier</td>
			<td>none</td>
			<td>12 weaks and male</td>
		</tr>
	</tbody>
</table>

an efficient and reproducible protocol for distraction osteogenesis in a rat model leading to a functional regenerated femur

本协议描述了在大鼠股骨模型中使用一种新研制的外固定器牵引成骨术。牵引成骨术 (做) 是一种外科手术技术, 导致骨再生后, 截切。osteotomized 肢通过逐渐的分散而相互移动, 达到预期的伸长率。该方法广泛应用于人下肢延长, 骨不连后治疗, 或骨肿瘤切除术后骨缺损的再生, 以及颌面部重建。只有少数的研究清楚地证明了他们的协议在获得功能性再生骨的效率,即, 在取出外固定架后, 将支持不带骨折的生理负重的骨骼。此外, 由于缺乏信息, 使得不同的研究和重复性的协议受到限制, 使学习比较困难。本研究的目的是制定一个可复制的协议, 包括一个适当的外固定器设计大鼠肢体延长, 与详细的手术技术, 允许动物的生理负重后, 去除外部固定.

从外科手术结束到合并结束的 x 线图像显示, 股骨 half-threaded 针没有松动, 表明稳定的锚固。别针是平行和保存完好。在执行过程中, osteotomized 肢沿骨骼的纵轴定向 (图 2)。在潜伏期结束时, 没有钙化区域是可见的 (图 2B)。在分心期结束时, 一些钙化区可见于先前存在的皮层 (图 2C)?...

Watch this Scientific Journal Video about An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur at JoVE.com

An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur

大鼠牵张成骨的高效可再生性协议

这项研究描述了一个可复制和详细的协议使用新开发的外固定器的牵引成骨 (做) 在股骨大鼠模型, 允许生理负重的动物后, 外固定架。

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) ...

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蛋白质和细胞坐月子创建二维图案衬底

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Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

Applications of In Vivo Functional Testing of the Rat Tibialis Anterior for Evaluating Tissue Engineered Skeletal Muscle Repair

中的应用<em&gt;在体内</em&gt;评定组织的鼠胫骨前的功能测试工程化骨骼肌修复

Despite the regenerative capacity of skeletal muscle, permanent functional and/or cosmetic deficits (e.g., volumetric muscle loss (VML) resulting from ...

A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation

基于数字图像相关的骨折检测中股骨皮质应变的估算方法

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral ...

An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

实验和有限元议定书调查中性和带电溶质的跨关节软骨运输

Osteoarthritis (OA) is a debilitating disease that is associated with degeneration of articular cartilage and subchondral bone. Degeneration of articular ...

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen

电化学 DNA 生物传感器检测食源性致病菌的研究进展

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, ...