Name: establishing a diaphyseal femur fracture model in mice
Uploaded: 2022-12-09T14:30:00
Description: Watch this Scientific Journal Video about Establishing a Diaphyseal Femur Fracture Model in Mice at JoVE.com

这项工作由里约热内卢州卡洛斯·查加斯·菲略研究支持基金会（FAPERJ）资助。

所有实验均已获得里约热内卢联邦大学健康科学中心动物使用和护理委员会的批准（协议编号101/21）。本研究使用10-12周龄（25-30g体重）的雄性Balb/c小鼠。每只小鼠的手术过程大约需要 15-20 分钟。在每次手术之前，所需的器械（列在 材料表中）必须组织在覆盖手术台的无菌手术区域（图1A）。金属手术器械必须在123°C的自密封信封中高压灭菌30分钟。一次性物品...

<ol>
	<li>Florencio-Silva, R., Sasso, G. R., Sasso-Cerri, E., Simoes, M. J., Cerri, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+of+bone+tissue:+Structure,+function,+and+factors+that+influence+bone+cells.">Biology of bone tissue: Structure, function, and factors that influence bone cells.</a> BioMed Research International. 2015, 421746 (2015).</li><li>Bahney, C. 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=Cellular+biology+of+fracture+healing.">Cellular biology of fracture healing.</a> Journal of Orthopedic Research. 37 (1), 35-50 (2019).</li><li>Einhorn, T. A., Gerstenfeld, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fracture+healing:+Mechanisms+and+interventions.">Fracture healing: Mechanisms and interventions.</a> Nature Reviews Rheumatology. 11 (1), 45-54 (2015).</li><li>Perren, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fracture+healing:+Fracture+healing+understood+as+the+result+of+a+fascinating+cascade+of+physical+and+biological+interactions.+Part+II.">Fracture healing: Fracture healing understood as the result of a fascinating cascade of physical and biological interactions. Part II.</a> Acta Chirurgiae Orthopaedicae et Traumatologiae Cechoslovaca. 82 (1), 13-21 (2015).</li><li>Giannoudis, P. V., Krettek, C., Lowenberg, D. W., Tosounidis, T., Borrelli, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fracture+healing+adjuncts-The+world's+perspective+on+what+works.">Fracture healing adjuncts-The world's perspective on what works.</a> Journal of Orthopaedic Trauma. 32, 43-47 (2018).</li><li>Kates, S. L., 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=Outside+the+bone:+What+is+happening+systemically+to+influence+fracture+healing.">Outside the bone: What is happening systemically to influence fracture healing.</a> Journal of Orthopaedic Trauma. 32, 33-36 (2018).</li><li>Ding, Z. C., Lin, Y. K., Gan, Y. K., Tang, T. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+pathogenesis+of+fracture+nonunion.">Molecular pathogenesis of fracture nonunion.</a> Journal of Orthopaedic Translation. (14), 45-56 (2018).</li><li>Calori, G. 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-unions.">Non-unions.</a> Clinical Cases in Mineral Bone Metabolism. 14 (2), 186-188 (2017).</li><li>Ekegren, C. L., Edwards, E. R., de Steiger, R., Gabbe, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence,+costs+and+predictors+of+non-union,+delayed+union+and+mal-union+following+long+bone+fracture.">Incidence, costs and predictors of non-union, delayed union and mal-union following long bone fracture.</a> Internation Journal of Environmental Research and Public Health. 15 (12), 2845 (2018).</li><li>Aziziyeh, R., 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+burden+of+osteoporosis+in+four+Latin+American+countries:+Brazil,+Mexico,+Colombia,+and+Argentina.">The burden of osteoporosis in four Latin American countries: Brazil, Mexico, Colombia, and Argentina.</a> Journal of Medical Economics. 22 (7), 638-644 (2019).</li><li>Kostenuik, P., Mirza, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fracture+healing+physiology+and+the+quest+for+therapies+for+delayed+healing+and+nonunion.">Fracture healing physiology and the quest for therapies for delayed healing and nonunion.</a> Journal of Orthopaedic Research. 35 (2), 213-223 (2017).</li><li>Gomez-Barrena, 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=fracture+healing:+cell+therapy+in+delayed+unions+and+nonunions.">fracture healing: cell therapy in delayed unions and nonunions.</a> Bone. 70, 93-101 (2015).</li><li>Schlundt, C., 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=Clinical+and+research+approaches+to+treat+non-union+fracture.">Clinical and research approaches to treat non-union fracture.</a> Current Osteoporosis Reports. 16 (2), 155-168 (2018).</li><li>Gomez-Barrena, 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=Feasibility+and+safety+of+treating+non-unions+in+tibia,+femur+and+humerus+with+autologous,+expanded,+bone+marrow-derived+mesenchymal+stromal+cells+associated+with+biphasic+calcium+phosphate+biomaterials+in+a+multicentric,+non-comparative+trial.">Feasibility and safety of treating non-unions in tibia, femur and humerus with autologous, expanded, bone marrow-derived mesenchymal stromal cells associated with biphasic calcium phosphate biomaterials in a multicentric, non-comparative trial.</a> Biomaterials. 196, 100-108 (2018).</li><li>Ryan, G., 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=Systemically+impaired+fracture+healing+in+small+animal+research:+A+review+of+fracture+repair+models.">Systemically impaired fracture healing in small animal research: A review of fracture repair models.</a> Journal of Orthopedic Research. 39 (7), 1359-1367 (2021).</li><li>Marmor, M. T., Dailey, H., Marcucio, R., Hunt, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomedical+research+models+in+the+science+of+fracture+healing+-+Pitfalls+&amp;+promises.">Biomedical research models in the science of fracture healing - Pitfalls &amp; promises.</a> Injury. 51 (10), 2118-2128 (2020).</li><li>Schindeler, A., Mills, R. J., Bobyn, J. D., Little, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+models+for+orthopedic+research+and+bone+tissue+engineering.">Preclinical models for orthopedic research and bone tissue engineering.</a> Journal of Orthopedic Research. 36 (3), 832-840 (2018).</li><li>Ewald, A. J., Werb, Z., Egeblad, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+of+vital+signs+for+long-term+survival+of+mice+under+anesthesia.">Monitoring of vital signs for long-term survival of mice under anesthesia.</a> Cold Spring Harbor Protocols. 2011 (2), 5563 (2011).</li><li>Stollings, L. 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=Immune+modulation+by+volatile+anesthetics.">Immune modulation by volatile anesthetics.</a> Anesthesiology. 125 (2), 399-411 (2016).</li><li>Sedghi, S., Kutscher, H. L., Davidson, B. A., Knight, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volatile+anesthetics+and+immunity.">Volatile anesthetics and immunity.</a> Immunological Investigations. 46 (8), 793-804 (2017).</li><li>Tsukamoto, A., Serizawa, K., Sato, R., Yamazaki, J., Inomata, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vital+signs+monitoring+during+injectable+and+inhalant+anesthesia+in+mice.">Vital signs monitoring during injectable and inhalant anesthesia in mice.</a> Experimental Animals. 64 (1), 57-64 (2015).</li><li>Kom&#225;rek, V., Hedrich, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+2.2.+Gross+anatomy.">Chapter 2.2. Gross anatomy.</a> The Laboratory Mouse (Second Edition). , 145-159 (2012).</li><li>Amend, S. R., Valkenburg, K. C., Pienta, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+hind+limb+long+bone+dissection+and+bone+marrow+isolation.">Murine hind limb long bone dissection and bone marrow isolation.</a> Journal of Visualized Experiments. (110), e53936 (2016).</li><li>An, Y. H., Moreira, P. L., Kang, Q. K., Gruber, H. E., An, Y. H., Martin, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+embedding+and+common+protocols.">Principles of embedding and common protocols.</a> Handbook of Histology Methods for Bone and Cartilage. , 185-197 (2003).</li><li>Enninghorst, N., McDougall, D., Evans, J. A., Sisak, K., Balogh, Z. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Population-based+epidemiology+of+femur+shaft+fractures.">Population-based epidemiology of femur shaft fractures.</a> Journal of Trauma and Acute Care Surgery. 74 (6), 1516-1520 (2013).</li><li>Gunderson, Z. J., Campbell, Z. R., McKinley, T. O., Natoli, R. M., Kacena, M. 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+comprehensive+review+of+mouse+diaphyseal+femur+fracture+models.">A comprehensive review of mouse diaphyseal femur fracture models.</a> Injury. 51 (7), 1439-1447 (2020).</li><li>Haffner-Luntzer, M., Fischer, V., Ignatius, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+fracture+healing+between+female+and+male+C57BL/6J+mice.">Differences in fracture healing between female and male C57BL/6J mice.</a> Frontiers in Physiology. 12, 712494 (2021).</li><li>Bonnarens, F., Einhorn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+a+standard+closed+fracture+in+laboratory+animal+bone.">Production of a standard closed fracture in laboratory animal bone.</a> Journal of Orthopaedic Research. 2 (1), 97-101 (1984).</li><li>Streubel, P. N., Desai, P., Suk, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+RIA+and+conventional+reamed+nailing+for+treatment+of+femur+shaft+fractures.">Comparison of RIA and conventional reamed nailing for treatment of femur shaft fractures.</a> Injury. 41, 51-56 (2010).</li></ol>

随着全球骨折数量的增加 9,10,25，愈合不愈合的创新治疗方法变得越来越紧迫。由于骨折愈合涉及对在很长一段时间内发生的事件的复杂而紧密的总结3，因此使用有效的动物模型对于提高我们对决定骨修复成功与否的机制的理解以及选择有效的药物和治疗方案至关重要16,17<sup class="xr...

骨骼是一个重要且功能多样的器官。骨骼的骨骼使身体姿势和运动成为可能，保护内脏器官，产生整合生理反应的激素，并且是造血和矿物质储存的场所1.如果骨折，骨骼具有非凡的再生能力，可以完全恢复其受伤前的形态和功能。愈合过程始于血肿和炎症反应的形成，这诱导骨膜、骨内膜和骨髓的骨骼干/祖细胞的活化和凝聚，以及随后的分化形成软骨愈伤组织。然后，断裂末端的桥接通过类似于软骨内骨形成的过程发生，其中软骨支架膨胀然后矿化，形成坚硬的骨性愈伤组织。最后，硬愈伤组织逐渐由破骨细胞和成骨细胞重塑，以恢复原始骨骼结构 2,3。
尽管骨折愈合过程相当稳健，但它涉及一系列错综复杂的事件，并受到几个个体因素的显着影响，包括患者的一般健康状况、年龄和性别，以及损伤因素，例如骨折的机械稳定方式、感染的发生以及周围软组织损伤的严重程度4，5,6.因此，失败很常见，导致骨不连的发展，这极大地影响了患者的康复和生活质量 7,8。由于高能量创伤和衰老导致的骨折数量增加，以及治疗费用高昂，不愈合骨折已成为全世界卫生系统的负担 9,10。这种日益增加的负担凸显了对创新治疗策略的迫切需要，以基于骨骼/间充质干细胞/基质细胞和仿生生物材料的组合来改善骨修复11,12 13,14。
为了实现这一目标，动物模型已被广泛用于旨在了解骨折愈合机制基础生物学的研究，以及旨在设计新的治疗策略以促进骨修复的概念验证临床前研究15,16,17。小动物模型，如小鼠，非常适合骨折愈合研究，因为转基因菌株和试剂可用于实验分析，而且维护成本低。此外，小鼠具有快速的愈合时间过程，这允许对修复过程的所有阶段进行时间分析15。然而，动物的小体型可能会给骨折的手术产生带来挑战，其固定模式类似于人类的固定模式。该协议描述了一种简单且低成本的小鼠骨折愈合模型，使用用髓内线稳定的开放式股骨截骨术，类似于最常见的骨修复过程，通过软骨痂的形成，并且可用于需要进入骨折部位的基础和转化研究。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Alcohol 70&#186;</td>
			<td>Merck</td>
			<td>109-56-8</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Canada balsam (mounting medium)</td>
			<td>Merck</td>
			<td>C1795</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Cefazoline</td>
			<td>ABL</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Merck</td>
			<td>CSL284525</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Dental X-Ray Generator</td>
			<td>Focus</td>
			<td>-</td>
			<td>Sold by Instrumentarium Dental Inc.&#160;</td>
		</tr>
		<tr>
			<td>DEPC water</td>
			<td>Merck</td>
			<td>W4502</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Dissecting Scissor</td>
			<td>ABC Instrumentos</td>
			<td>0327</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Vetec</td>
			<td>60REAVET014340</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Eosin solution</td>
			<td>Laborclin</td>
			<td>EA-65</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Ethanol P.A</td>
			<td>Vetec</td>
			<td>60REAVET012053</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Gauze&#160;pads</td>
			<td>Cremer</td>
			<td>Not applicable</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Harris Hematoxylin Solution</td>
			<td>Laborclin</td>
			<td>620503</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Tonkey Electrical Technology</td>
			<td>E114273</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Histological slides</td>
			<td>Merck</td>
			<td>CSL294875X25</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Histology cassettes</td>
			<td>Merck</td>
			<td>H0542-1CS</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Hydrochloric acid - 37%</td>
			<td>Merck</td>
			<td>258148</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Insulin syringe</td>
			<td>BD</td>
			<td>324918</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Iodopovidone&#160;sponge</td>
			<td>Rioqu&#237;mica</td>
			<td>372106</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Ketamine hydrochloride</td>
			<td>Ceva</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Lacribel collyrium</td>
			<td>Cristalia</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Leica</td>
			<td>149AUTO00C1</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse Tooth Forceps Tweezer</td>
			<td>ABC Instrumentos</td>
			<td>0164</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Needle 26 G</td>
			<td>BD</td>
			<td>2239</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Needle Holder&#160;</td>
			<td>Golgran</td>
			<td>135-18</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Nonresorbable Nylon Suture thread n&#186; 6</td>
			<td>Atramat</td>
			<td>C1546-NT</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Paraffin</td>
			<td>Exodo</td>
			<td>8002 - 74 - 2</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Sigma</td>
			<td>30525-89-4</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>PBS 1x&#160;</td>
			<td>Lonza</td>
			<td>&#160;BE17-516F</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Resorbable Nylon Suture thread n&#186; 6</td>
			<td>Atramat</td>
			<td>C1596-45B</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Rod Wire SS CrNi 0.016&#34;</td>
			<td>Orthometric</td>
			<td>56.50.2016</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel n&#186; 11</td>
			<td>Descarpak</td>
			<td>15782</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Serrated Tip Tweezer</td>
			<td>Quinelato</td>
			<td>QC.404.12</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Phillips</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Surgical tape</td>
			<td>3M</td>
			<td>2734</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Surgical tnt field</td>
			<td>Polarfix</td>
			<td>6153</td>
			<td>Or any general available supplier</td>
		</tr>
		<tr>
			<td>Tramadol hydrochloride</td>
			<td>Teuto&#160;</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Water bath for histology</td>
			<td>Leica</td>
			<td>HI1210</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine hydrochloride</td>
			<td>Ceva</td>
			<td>Not applicable</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Dinamica</td>
			<td>60READIN001105</td>
			<td>Similar brands of the item may be used according to local availability</td>
		</tr>
	</tbody>
</table>

establishing a diaphyseal femur fracture model in mice

骨骼具有显着的再生能力。然而，骨折愈合是一个复杂的过程，根据病变的严重程度以及患者的年龄和整体健康状况，可能会发生失败，导致愈合延迟或不愈合。由于高能创伤和衰老导致的骨折数量不断增加，迫切需要开发基于骨骼/间充质干细胞/基质细胞和仿生生物材料相结合的创新治疗策略来改善骨修复。为此，使用可靠的动物模型对于更好地了解决定愈合结果的关键细胞和分子机制至关重要。在所有模型中，小鼠是首选的研究模型，因为它提供了多种用于实验分析的转基因菌株和试剂。然而，由于小鼠体型小，在小鼠中建立骨折可能在技术上具有挑战性。因此，本文旨在展示通过软骨愈伤组织形成的小鼠骨干股骨骨折的手术建立程序，该骨折由髓内线稳定，类似于最常见的骨修复过程。

评估外科手术在骨折方面是否成功的最简单和最直接的方法是 X 射线成像。可以在手术后立即进行 X 光片，小鼠仍处于麻醉状态，随后在骨折后 7 天、14 天和 21 天进行 X 光检查，以评估愈伤组织形成和进展。可接受的骨折模式是皮质完全破裂，钢丝正确放置在髓管内，并且骨折线是横向的（与骨轴成 90° 角）、斜的（弯曲或倾斜的模式，没有碎片位移）或短斜（相对于骨轴约 30°）（<strong class="x...

Watch this Scientific Journal Video about Establishing a Diaphyseal Femur Fracture Model in Mice at JoVE.com

Establishing a Diaphyseal Femur Fracture Model in Mice

该协议描述了一种外科手术，用于在小鼠股骨中建立骨干骨折，该骨折用髓内线稳定，用于骨折愈合研究。

建立小鼠骨干股骨骨折模型

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age ...

这里展示的骨干股骨骨折模型是一种简单且低成本的小鼠模型，适用于长骨骨折愈合的基础和临床前研究。开放手术技术不需要复杂的手术器械和固定装置。它允许目视检查正在产生的裂缝。该模型可用于局部移植和治疗成分的体内测试，包括无法通过经皮或全身注射递送的干细胞、药物和生物材料。手术方案需要一些基本手术技术和小鼠解剖学知识才能执行关键步骤。这项工作中的视觉演示甚至可以帮助那些没有动物手术经验的人。莱昂纳多·穆勒（Leonardo Muller）和比安卡·弗拉德（Bianca Frade）将演示该程序，他们都是干细胞和骨修复实验室的硕士生。将麻醉的动物转移到手术台后，用10%聚维酮碘海绵擦拭皮肤，对要切开的区域进行消毒。然后用无菌纱布垫擦干摩擦区域，用70%乙醇洗涤，然后用无菌纱布垫再次干燥。接下来，将鼠标置于右侧侧卧位并悬垂鼠标，仅显示切口区域。手术前，将眼药水滴入小鼠的眼睛以避免干燥，并在手术过程中不断检查鼠标的呼吸。用 11 号手术刀刀片做一个 1 厘米的皮肤髌旁外侧切口，从胫骨粗隆水平开始，延伸到髌骨水平，然后向股骨远端保持相等的距离。用钝头剪刀解剖切口线周围的皮下筋膜，露出阔筋膜、股外侧肌和股二头肌。用 11 号手术刀刀片，在阔筋膜上再做一个切口，类似于在皮肤上做的切口，从胫骨粗隆水平开始，沿着股二头肌腱膜延伸，直到股骨远端水平打开关节囊并进入膝关节。将直锯齿状精密尖端镊子的尖端放在髌骨下方，并将其与髌骨和股四头肌韧带一起推到一边，从而暴露股骨髁突，从而进行髌骨内侧脱位。用锯齿状尖端镊子握住股骨，将膝盖弯曲 90 度，然后用 26 号皮下注射针通过髁间窝手动穿孔股骨髓内管。保持膝盖弯曲 90 度，将一段 1.0 厘米的 0.016 英寸不锈钢棒线穿过开口进入股骨髓管，朝向大转子。用直锯齿状尖端镊子调整导线的远端，将其紧紧地固定在外侧髁中。接下来，使用锯齿状尖端镊子通过钝端解剖分离股外侧股二头肌和股二头肌，以进入股骨远端骨干。将解剖剪刀以大约 90 度的角度插入股骨骨干周围，然后轻轻地进行完整的皮质截骨术。通过将直锯齿状精密尖端镊子的尖端推到髁突区域来重新定位肌肉和髌骨。用 6-0 可重吸收缝合线闭合肌肉筋膜，然后使用 6-0 尼龙缝合线闭合皮肤，两者都以简单的中断方式进行。可接受骨折模式的 X 线片显示横向骨干骨折，其中骨折线与骨轴成 90 度角，骨折线相对于骨轴小于 30 度的短斜骨折，可复位的碎片性骨折，可以看到一些小的骨碎片， 但骨骼的解剖学排列仍然存在。这里显示了错误放置的钢丝的代表性 X 光片，其中钢丝不在股骨近端碎片的髓管内，从而导致骨折骨的不正确固定，以及钢丝没有穿过任何骨碎片并且骨折骨完全未对齐的情况。术后第 14 天和第 21 天骨折部位可见愈伤组织表明模型的再生过程。组织学分析显示骨折部位愈伤组织的评估。骨胼痂最初在第 7 天出现，在骨折线周围呈现出大面积的高线软骨区域。在第 14 天，在软骨区域周围观察到骨化前沿，形成小梁骨和充满重建骨髓的空腔。最后，在第 21 天，软骨区域几乎完全被小梁骨取代，表明骨桥接成功。模型的成功取决于导线的正确插入。因此，在此步骤中保持膝盖弯曲 90 度角至关重要。此外，微观计算机人口统计学可用于评估细胞的进化和在三维和定量尺度上形成的骨骼的特征。

Research

JoVE Journal

Biology

Intravitreous Injection for Establishing Ocular Diseases Model

玻璃体内注射眼部疾病模型建立

Intravitreous injection is a widely used technique in visual sciences research. It can be used to establish animal models with ocular diseases or as ...

科研

生物学

An Alternant Method to the Traditional NASA Hindlimb Unloading Model in Mice

传统美国宇航局后肢卸小鼠模型的一个交替的方法

The Morey-Holton hindlimb unloading (HU) method is a widely accepted National Aeronautics and Space Administration (NASA) ground-based model for studying ...

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the ...

Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy

冷冻断裂/冷冻蚀刻的生物电子显微技术的基本要素

Freeze-fracture/freeze-etch describes a process whereby specimens, typically biological or nanomaterial in nature, are frozen, fractured, and replicated ...

Validation of a Mouse Model to Disrupt LINC Complexes in a Cell-specific Manner

小鼠模型的验证扰乱LINC配合物在细胞特异性的方式

Nuclear migration and anchorage within developing and adult tissues relies heavily upon large macromolecular protein assemblies called LInkers of the ...

The Assembly and Application of 'Shear Rings': A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress

大会和“剪切环”中的应用：一种新型内皮模型轨道，单向和定期流体流动和剪应力

Deviations from normal levels and patterns of vascular fluid shear play important roles in vascular physiology and pathophysiology by inducing adaptive as ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

质谱分析，用于识别 EYFP 标记 CENP-A （EYFP-CENP-A） 的泛化

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

A Capsule-Based Model for Immature Hard Tick Stages Infestation on Laboratory Mice

基于胶囊的模型，用于在实验鼠身上的不成熟硬滴答阶段侵扰

Ticks are obligatory blood feeding parasites at all stages of development (except eggs) and are recognized as vectors of various pathogens. The use of ...

A Modified Surgical Model of Hind Limb Ischemia in ApoE-/- Mice using a Miniature Incision

A Modified Surgical Model of Hind Limb Ischemia in ApoE-/- Mice using a Miniature Incision

ApoE小鼠后肢缺血的改进手术模型-/- 使用微型切口

The purpose of this study is to introduce and evaluate a modified surgical approach to induce acute ischemia in mice that can be implemented in most ...

Tail Vein Transection Bleeding Model in Fully Anesthetized Hemophilia A Mice

全麻醉血友病A小鼠尾静脉横断出血模型

Tail bleeding models are important tools in hemophilia research, specifically for the assessment of procoagulant effects. The tail vein transection (TVT) ...