Bu eser RISystem AG, Davos, İsviçre tarafından desteklenmiştir.

Tüm yordamlar deneysel hayvan kullanımı için Ulusal Sağlık Enstitüleri esaslarına göre gerçekleştirilen ve kurumsal yönergeleri (Landesamt für Verbraucherschutz, Zentralstelle Amtstierärztlicher Dienst, Saarbrücken, takip Almanya).
1. cerrahi aletler ve implantlar hazırlanması
<ol><li>Bir neşter blade (mevcudu 15), küçük bir bez, iyi forseps, 27 G iğne, tekrar Emilebilen olmayan 5-0 dikiş, makas ve bir iğne bağı mikrocerrahi araç kutusundan seçin.</li>
	<li>İntramedüller vida açmak kılavuz tel (0.3/0.2 mm çapında, 10 cm uzunluk), merkezleme matkap (0.5 mm çapında) ve el matkap (şekil 1; bkz: Malzemeler tablo). Not: İntramedüller vida (0.5 mm çap, 17.2 mm Uzunluk) uyluk kemiği içine retrograd implantasyon için Medikal paslanmaz çelikten yapılmıştır. Vida bir proksimal iş parçacığı (0.5 mm çaplı, 4 mm Uzunluk) bir burun (0.2 mm çap, 0.4 mm Uzunluk) ile uç ve kırık sıkıştırma ve eksenel ve dönme istikrar ulaşmak için distal koni şeklinde kafa (0.8 mm çap, 0.9 mm Uzunluk) vardır.</li>
	<li>İmplantlar ve 5 min için dezenfeksiyon çözüm (% 96'sı alkol) için tüm cerrahi aletler bulunabileceği veya onları (buharlı sterilizasyon, 130 ° C, 25 dk) sterilize. Dezenfeksiyon veya sterilizasyon sonra aletleri üzerinde bir işlem bez koyun. Küçük hayvan işlemi tabloya bitişik çalışma bez getirin.</li>
</ol>2. hayvan, anestezi ve Analjezi
<ol><li>Zorlanma, yaş ve cinsiyet yöneliktir çalışma soru göre farelerin seçin. Not: Bu çalışma için 12 - için 14-hafta-yaşlı erkek CD-1 fareler kullanıldı. İntramedüller vida kullanmak için uygun vücut ağırlığı arasında 25-35 g olduğunu.</li>
	<li>Mayi enjeksiyonu 15 mg/kg xylazine ve 75 mg/kg ketamin ile fareler anestezi. Anesthetization ayak çimdik tarafından onaylayın. Anestezi sırasında kurumasını hayvanların gözleri korumak için göz yağ uygulayın. Anestezi indüksiyon sonra vücut ısısını sabit tutmak için bir ısı radyatör altında fareyi getirin. İşlem sırasında hayvanları anestezi uygun bir uçak emin olmak için tekrar tekrar ayak çimdik ile takip.</li>
	<li>Tramadol-hidroklorid (1,0 mg/mL) içme suyundaki analjezi için ameliyattan sonra 3 güne kadar ameliyattan önce 1 gün uygulayın. Not: Analjezi ve Enfeksiyon önleme deneyler yapılması için nerede, ülke ve kurum ilgili yönergeleri ile anlaşma olması gerekir.</li>
</ol>3. cerrahi müdahale ve intramedüller vida implantasyon
<ol><li>Ameliyatından önceki tüm sağ arka bacak tıraş ve depilatory bir krem uygulayın. 5 dk sonra krem çıkarın ve bacak su ile temizleyin. Daha sonra % 96'sı alkol ile dezenfeksiyon bir çözüm uygulayın. Betadine veya klorheksidin tam asepsis emin olmak için alkol ilave edilebilir.</li>
	<li>Aseptik koşullar altında küçük hayvan işlemi tablosundaki sırtüstü pozisyonda fareyi getirin. Anterior yaklaşım diz condyles için izin vermek için doğru dizini kır. Neşter bıçak kullanarak sağ diz, 5 mm medial parapatellar kesi gerçekleştirin.</li>
	<li>Dikkatle neşter blade ve pamuklu çubuk ile Patella bağ seferber. O zaman, patella yanal femur intercondylar çentik ortaya çıkarmak için iyi Forseps ile vardiya.</li>
	<li>Tam ortasında her iki condyles arasında uyluk intercondylar çentik açın. 1.0 mm matkap delik için derinlemesine aşmayacak şekilde emin olun.<ol>
<li>A yavaş hız ve 45 ° ofset ventrally 0,5 mm matkap ve el matkap (şekil 1 c ve D, Şekil 2) merkezleme kullanarak uyluk eksenine sondaj el ile başlatın. Delme sırasında sürekli açı 0 ° uzaklığı (femur kemik eksen ile paralel) azaltın. 1.0 mm derinliğe ulaştığında sondaj durdurmak.</li>
	</ol>
</li>
	<li>Kemikte intercondylar çentik açtıktan sonra 27 G iğne üzerinde femur uzunluğu tüm intramedüller boşluğa yerleştirmek. 27 G iğne döner hareketleri el ile uyluk intramedüller boşluğuna ream. Büyük trochanter kortikal kemikte proksimale ekleyip için iğne ileri itin.</li>
	<li>27 G iğne kaldırmak ve femur distal parçası Kılavuzu tellerden uygulayın.<ol>
<li>Bir cilt kesi ile bir neşter bıçak (mevcudu 15) proksimale kılavuz tel üzerinden yapmak ve her iki ucunda da Kılavuzu dışında olana Kılavuzu ileri itin. Kılavuzu yerinde tutmak emin olun.</li>
	</ol>
</li>
	<li>Tanımlanmış bir kapalı kırık Giyotin kullanarak oluşturun.<ol>
<li>Giyotin altında sağ bacak ile lateral pozisyonda fareyi getirin. Diaphyseal kısmı femur ortasında Giyotin yerleştirildiğinden emin olun.</li>
		<li>Ağırlık (200 gr) tanımlanmış 25.5 cm mesafe bırakın.</li>
	</ol>
</li>
	<li>X-ray cihazı kullanarak kılavuz tel (şekil 3) konumunu yanı sıra kırık yapılandırma ve kırık pozisyon kontrol ( Tablo malzemelerigörmek).</li>
	<li>Distal uçta burun 0.2 mm kılavuz tel intramedüller vida bağlanabildiğinizi ve uyluk sürekli basınç ve saat yönünde döndür altında takın.<ol>
<li>Ne zaman yeterli tork elde mili makas.</li>
		<li>Kılavuzu proksimale kaldırın.</li>
	</ol>
</li>
	<li>Diz kapağı yeniden konumlandırmak ve kaslara patella tendon bir tek dikiş kullanarak bir 5-0 sentetik, monofilament, nonabsorbable polipropilen sütür ile düzeltin. Aynı malzeme ve boyutu tek dikiş Yarayı kapatmak için kullanın. Parçaları ve radiologically x-ray cihazı kullanarak vidalı konum azaltılması kontrol ( Tablo malzemelerigörmek).</li>
	<li>Anesteziden kurtarmak kadar hayvanlar ısı radyatör altında tutun. Onlar ventral recumbency korumak için yeterli bilinci yerine kadar hayvanları sahipsiz bırakmayın. Hayvanlar hayvan tesisi tek kafeslerde dönün. Onlar tamamen anesteziden iyileşti olsa bile hayvanlar ilk 24 saat sırasında diğer hayvanlar şirkete iade etmeyin.</li>
	<li>Hayvanlar her gün dikkatle izleyin. Ameliyat sonrası analjezi tramadol-hidroklorid 1.0 mg/mL bir doz ile içme suyu ilk üç gün boyunca kullanarak koruyun. Gün 4 ameliyat sonrası, hayvanlar hala ağrı, kanıt göstermek, Analjezi seslendirme, huzursuzluk, hareketlilik, damat için başarısızlık, anormal Postür ve çevresi normal ilgi eksikliği eksikliği tarafından belirtildiği şekilde devam. Hayvanlar olduğunda analjezi sonlandırmak ağrısız.</li>
	<li>Deney sonunda hayvan aşırı doz barbitürat tarafından ötenazi.</li>
</ol>

<ol>
	<li>Jacenko, O., Olsen, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+mouse+models+in+studies+of+skeletal+disorders.">Transgenic mouse models in studies of skeletal disorders.</a> J Rheumatol Suppl. 43, 39-41 (1995).</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=Small+animal+bone+healing+models:+standards,+tips,+and+pitfalls+results+of+a+consensus+meeting.">Small animal bone healing models: standards, tips, and pitfalls results of a consensus meeting.</a> Bone. 49 (4), 591-599 (2011).</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> J Orthop Res. 2 (1), 97-101 (1984).</li><li>Holstein, J. H., 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=Development+of+a+stable+closed+femoral+fracture+model+in+mice.">Development of a stable closed femoral fracture model in mice.</a> J Surg Res. 153 (1), 71-75 (2009).</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=Ex+vivo+analysis+of+rotational+stiffness+of+different+osteosynthesis+techniques+in+mouse+femur+fracture.">Ex vivo analysis of rotational stiffness of different osteosynthesis techniques in mouse femur fracture.</a> J Orthop Res. 27 (9), 1152-1156 (2009).</li><li>Claes, L., Augat, P., Suger, G., Wilke, 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=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>

Yazarlar onlar rakip hiçbir mali çıkarları var bildirin.

Kritik adımlardır cerrahi prosedürün vida implantasyon uyluk condyles intercondylar çentik hem de iğne en iyi yönünü ortasında, oyma için kemik eksenine paralel doğru giriş noktası bulmak için intramedüller kavite. Cerrah bir hatalı giriş pozisyon önlemek için en iyi görünümü elde kadar çentik hazır olmalıyız. Raybalamada sırasında yönünü denetlemek için fareyi uyluk istikrarlı bir pozisyonda parmaklarıyla tutulmalıdır. Kılavuzu bir kırık çıkık kaynaklanan proksimal kemik parç...

Farelerde kemik şifa çalışmalar antikorlar ve hayvanlar genetik olarak değiştirilmiş geniş bir yelpazede nedeniyle büyük talep vardır. Bu gerçekleri1şifa kemik moleküler mekanizmaları incelemek için izin verir. Son birkaç yıl içinde2farklı kemik fareler olmuştur için modelleri şifa geliştirdi. Açık modelleri, hangi kemik açık bir yanal cerrahi yaklaşım kullanarak osteotomized ve Bonnares ve Einhorn3tarafından tanıtıldı kırık modelini temel kemik kırılmış kapalı modelleri bu modeller ayrılabilir. Bu tekniği kullanarak, standart bir transvers kırık 3 noktalı bükme aygıt tarafından üretilebilir ve intramedüller implantlar küçük medial parapatellar kesi büyük yumuşak doku travması kaçınarak bir minimal invaziv tekniği yoluyla eklenebilir.
İntramedüller vida kapalı kırık sabitleme farelerde için uygulanabilir. Vida dönme ve eksenel istikrar sunmaktadır. Bu kırık sıkıştırma iplik proksimal ve distal baş4aracılığıyla elde edilir. Vida basit cerrahi tekniği daha da avantajları, invasivity, düşük düşük dereceli implant ağırlık ve en önemlisi, daha yüksek bir kararlılık diğer intramedüller göre standart ve kontrollü biyomekanik koşulları sağlayan 5implantları. Aslında, en kapalı kırık modellerinde parçaları dönme ve eksenel istikrar tam bir eksikliği ve PIN yüksek riski ile ilişkili yalnızca basit iğne tarafından stabilize ve aynı zamanda Kırık çıkık. Bu belirgin gecikmeli şifa veya sendikasız oluşumuna neden olabilir iyileşme süreci etkileyebilir.
Tanınmış kırık fiksasyonu kararlılığını iyileşme süreci6,7üzerinde büyük bir etkisi vardır. Endochondral şifa micromovements kırık boşluğu içinde izin verebilir, daha yumuşak bir fiksasyon sonuçları yüksek rijit fiksasyonu Intramembranous iyileşmesinde, sonuçları. İstikrar kırığı intramedüller vida ile ağırlıklı olarak bir endochondral çok fazla Kallus dokusunun özellikle 2 kırık iyileşmesi haftalar süren iyileşme gösterir. Olasılığı çok fazla Kallus dokusunun hasat için birden çok parametre analizi tarafından farklı teknikler sağlar.
Burada, tasarım ve uygulama intramedüller vida farelerde yanı sıra avantajları ve dezavantajları normal endochondral kemik iyileşmesi deneysel çalışmalarda raporu.

<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 &amp;amp;CoKG</td><td>BD520R</td><td></td></tr><tr><td>Scissors</td><td>Braun Aesculap AG &amp;amp;CoKG</td><td>BC100R</td><td></td></tr><tr><td>Needle holder</td><td>Braun Aesculap AG &amp;amp;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 &amp;amp;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&amp;uuml;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

Kemik şifa modelleri klinik kırık tedavi geliştirmek için şifa kırık karmaşık mekanizmaları analiz etmek gereklidir. Çünkü fare modelleri genetiği değiştirilmiş suşları ve kırık iyileşmesi, moleküler mekanizmaları analizi için özel antikorlar, çok sayıda teklif son on yılda, fare modelleri ortopedik araştırma kullanımının artmasıyla, en büyük olasılıkla belirtilmişti. Biyomekanik koşulları denetlemek için iyi karakterize Osteosentez teknikleri de farelerde, zorunludur. Burada, tasarım ve kullanımı modeli şifa kapalı kemik ile uyluk kemiği kırıkları farelerde stabilize etmek için ilgili raporu. Medikal paslanmaz çelikten intramedüller vida kırık sıkıştırma Aksiyel ve dönme istikrar tam bir eksikliği göstermek çoğunlukla kullanılan basit intramedüller iğne karşılaştırıldığında bir Aksiyel ve dönme istikrar sağlar. İntramedüller vida ile elde istikrar endochondral şifa analiz sağlar. Sabitleme vida ile sonra alınan Kallus dokusunun büyük bir miktar doku biyokimyasal ve moleküler analizleri hasat için ideal koşulları sağlamaktadır. Vida kullanımı daha fazla avantajı vida minimal invaziv tekniği ile uyluk kemiği içine yumuşak doku hasar inducing olmadan eklenebilir bu gerçektir. Sonuç olarak, vida kapalı kırık standart biyomekanik koşullar sunan modelleri Şifa ideal olarak kullanılabilir benzersiz bir implantı olduğunu.

Kapatma yaralamaya cilt kesi gelen çalışma süresi 20 dk oldu. Stereo mikroskop ameliyat. Postoperatively, hayvanlar her gün takip. Hayvanların hiçbiri bu süre sonunda acı kanıtı gösterdi çünkü ameliyat sonrası analjezi 2 gün sonra sonlandırıldı. Hayvanlar da ameliyattan sonra 2 gün içinde normal ağırlık taşıyan gösterdi. Yara enfeksiyonları tüm gözlem döneminde gözlenen değil.
Radyolojik analizl...

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

Bu iletişim kuralı bir intramedüller vida farelerde şifa endochondral kemik çözümlemek için kullanılan uyluk kemiği kırıkları standart sabitleme için bir minimal invaziv Osteosentez tekniği açıklar.

Endochondral standart biyomekanik koşullar altında farelerde şifa kırık çözümlemek için minimal invaziv bir Model

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

a-minimally-invasive-model-to-analyze-endochondral-fracture-healing

Research

JoVE Journal

Medicine

8.2K Görüntüleme.  Saarland University. Bu iletişim kuralı bir intramedüller vida farelerde şifa endochondral kemik çözümlemek için kullanılan uyluk kemiği kırıkları standart sabitleme için bir minimal invaziv Osteosentez tekniği açıklar.

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

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Farelerde Basit Kritik ölçekli Femoral Hata Modeli

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Tıp

Establishment of a Segmental Femoral Critical-size Defect Model in Mice Stabilized by Plate Osteosynthesis

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a model organism, the mouse has already been widely used in bone-related research. Large diaphyseal bone defect models in mice, however, are sparse and often use bone fixation which fills the bone marrow cavity and does not provide optimal mechanical stability. The objectives of the current study were to develop a critical-size, segmental, femoral defect in nude mice. A 3.5-mm mid-diaphyseal femoral ostectomy (approximately 25% of the femur length) was performed using a dedicated jig, and was stabilized with an anterior located locking plate and 4 locking screws. The bone defect was subsequently either left empty or filled with a bone substitute (syngenic bone graft or coralline scaffold). Bone healing was monitored noninvasively using radiography and in vivo micro-computed-tomography and was subsequently assessed by ex vivo micro-computed-tomography and undecalcified histology after animal sacrifice, 10 weeks postoperatively. The recovery of all mice was excellent, a full-weight-bearing was observed within one day following the surgical procedure. Furthermore, stable bone fixation and consistent fixation of the implanted materials were achieved in all animals tested throughout the study. When the bone defects were left empty, non-union was consistently obtained. In contrast, when the bone defects were filled with syngenic bone grafts, bone union was always observed. When the bone defects were filled with coralline scaffolds, newly-formed bone was observed in the interface between bone resection edges and the scaffold, as well as within a short distance within the scaffold. 
The present model describes a reproducible critical-size femoral defect stabilized by plate osteosynthesis with low morbidity in mice. The new load-bearing segmental bone defect model could be useful for studying the underlying mechanisms in bone regeneration pertinent to orthopaedic applications.

Although mouse models are invaluable tools for bone tissue engineering, models of long bone defects are sparse. This need motivated development of the present protocol which uses a locking plate with four screws and a dedicated jig to perform and stabilize a reproducible, femoral, critical-size defect with low morbidity.

Farelerde bir Segmental Femoral Kritik boyutlu Defekt Modelinde kurulması Plaka Osteosentez tarafından Stabilize

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a ...

Real-time Visualization and Analysis of Chondrocyte Injury Due to Mechanical Loading in Fully Intact Murine Cartilage Explants

Homeostasis of articular cartilage depends on the viability of resident cells (chondrocytes). Unfortunately, mechanical trauma can induce widespread chondrocyte death, potentially leading to irreversible breakdown of the joint and the onset of osteoarthritis. Additionally, maintenance of chondrocyte viability is important in osteochondral graft procedures for optimal surgical outcomes. We present a method to assess the spatial extent of cell injury/death on the articular surface of intact murine synovial joints after application of controlled mechanical loads or impacts. This method can be used in comparative studies to investigate the effects of different mechanical loading regimens, different environmental conditions or genetic manipulations, as well as different stages of cartilage degeneration on short- and/or long-term vulnerability of in situ articular chondrocytes. The goal of the protocol introduced in the manuscript is to assess the spatial extent of cell injury/death on the articular surface of murine synovial joints. Importantly, this method enables testing on fully intact cartilage without compromising native boundary conditions. Moreover, it allows for real-time visualization of vitally stained articular chondrocytes and single image-based analysis of cell injury induced by application of controlled static and impact loading regimens. Our representative results demonstrate that in healthy cartilage explants, the spatial extent of cell injury depends sensitively on load magnitude and impact intensity. Our method can be easily adapted to investigate the effects of different mechanical loading regimens, different environmental conditions or different genetic manipulations on the mechanical vulnerability of in situ articular chondrocytes.

We present a method to assess the spatial extent of cell injury/death on the articular surface of intact murine joints after application of controlled mechanical loads or impacts. This method can be used to investigate how osteoarthritis, genetic factors and/or different loading regimens affect the vulnerability of in situ chondrocytes.

Gerçek zamanlı görselleştirme ve analiz nedeniyle tam olarak olduğu gibi fare kıkırdak Explants mekanik yükleme kondrosit yaralanma

Homeostasis of articular cartilage depends on the viability of resident cells (chondrocytes). Unfortunately, mechanical trauma can induce widespread ...

Biyomühendislik

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

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming more commonly used in trauma research. They offer a large number of mutant strains and antibodies for the analysis of the molecular mechanisms behind the highly differentiated process of bone healing. To control the biomechanical environment, standardized and well-characterized osteosynthesis techniques are mandatory in mice. Here, we report on the design and use of an intramedullary nail to stabilize open femur osteotomies in mice. The nail, made of medical-grade stainless steel, provides high axial and rotational stiffness. The implant further allows the creation of defined, constant osteotomy gap sizes from 0.00 mm to 2.00 mm. Intramedullary locking nail stabilization of femur osteotomies with gap sizes of 0.00 mm and 0.25 mm result in adequate bone healing through endochondral and intramembranous ossification. Stabilization of femur osteotomies with a gap size of 2.00 mm results in atrophic non-union. Thus, the intramedullary locking nail can be used in healing and non-healing models. A further advantage of the use of the nail compared to other open bone healing models is the possibility to adequately fix bone substitutes and scaffolds in order to study the process of osseous integration. A disadvantage of the use of the intramedullary nail is the more invasive surgical procedure, inherent to all open procedures compared to closed models. A further disadvantage may be the induction of some damage to the intramedullary cavity, inherent to all intramedullary stabilization techniques compared to extramedullary stabilization procedures.

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

Femur Osteotomiler standardize Sabitleme için bir intramedüller Kilitleme Nail Fare Normal ve Arızalı Kemik Şifa Analiz

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

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

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

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

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

Vivo Fare uyluk için bir MRI uyumlu Osteosentez aygıtı kullanma farelerde kemik iyileşme sırasında kırık Kallus gelişimi değerlendirme

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

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes. Here, we introduce a clinically relevant mouse model of tibial fracture. The postoperative changes in these mice mimic some of the cognitive impairments commonly observed after routine orthopedic surgery in humans. Briefly, an incision is performed in the right hind limb under strictly aseptic conditions. Muscles are disassociated, and a 0.38-mm stainless steel pin is inserted into the upper crest of the tibia, inside the intramedullary canal. Osteotomy is then performed, and the wound is stapled. We have used this model to investigate the effects of surgical trauma on postoperative neuroinflammation and behavioral changes. By applying this fracture model in combination with parabiosis, a surgical model in which 2 mice are anastomosed, we have studied cells and secreted factors that systemically rejuvenate organ function and tissue regeneration after injury. By following our step-by-step protocol, these models can be reproduced with high fidelity, and can be adapted to interrogate many biologic pathways that are altered by surgical trauma.

This protocol describes a mouse model of orthopedic surgery that has been used to study mechanisms of postoperative neuroinflammation and behavioral changes, and when combined with parabiosis, to study tissue regeneration during aging.

Ameliyat sonrası bilişsel işlev bozukluğu ve doku rejenerasyonu çalışmaya ortopedik cerrahi bir fare modeli

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies that promote efficient bone healing are non-existent and in high demand. Effective and reproducible animal models of fractures healing are needed to understand the complex biological processes associated with bone regeneration. Many animal models of fracture healing have been generated over the years; however, murine fracture models have recently emerged as powerful tools to study bone healing. A variety of open and closed models have been developed, but the closed femoral fracture model stands out as a simple method for generating rapid and reproducible results in a physiologically relevant manner. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice and facilitate a post-fracture stabilization of the femur by inserting an intramedullary steel rod. Although devices such as a nail or a screw offer greater axial and rotational stability, the use of an intramedullary rod provides a sufficient stabilization for consistent healing outcomes without producing new defects in the bone tissue or damaging nearby soft tissue. Radiographic imaging is used to monitor the progression of callus formation, bony union, and subsequent remodeling of the bony callus. Bone healing outcomes are typically associated with the strength of the healed bone and measured with torsional testing. Still, understanding the early cellular and molecular events associated with fracture repair is critical in the study of bone tissue regeneration. The closed femoral fracture model in mice with intramedullary fixation serves as an attractive platform to study bone fracture healing and evaluate therapeutic strategies to accelerate healing.

The murine closed femoral fracture model is a powerful platform to study fracture healing and novel therapeutic strategies to accelerate bone regeneration. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice using an intramedullary steel rod to stabilize the femur.

Farelerde kapalı femur kırığı nesil: kemik şifa çalışması için bir Model

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

Transverse Fracture of the Mouse Femur with Stabilizing Pin

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test whether a gene, gene product or drug influences bone repair because murine bones recapitulate the stages observed during human fracture healing. When a mouse or human breaks a bone, an inflammatory response is initiated, and the periosteum, a stem cell niche surrounding the bone itself, is activated and expands. Cells residing in the periosteum then differentiate to form a vascularized soft callus. The transition from the soft callus to a hard callus occurs as the recruited skeletal progenitor cells differentiate into mineralizing cells, and the bridging of the fractured ends results in the bone union. The mineralized callus then undergoes remodeling to restore the original shape and structure of the healed bone. Fracture healing has been studied in mice using various injury models. Still, the best way to recapitulate this entire biological process is to break through the cross-section of a long bone that encompasses both cortices. This protocol describes how a stabilized, transverse femur fracture can be safely performed to assess healing in adult mice. A surgical protocol including detailed harvesting and imaging techniques to characterize the different stages of fracture healing is also provided.

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

Fare Femurunun Stabilize Pin ile Enine Kırılması

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test ...

Biyoloji

Establishing a Diaphyseal Femur Fracture Model in Mice

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age and overall health status of the patient, failures can occur, leading to delayed union or nonunion. Due to the increasing number of fractures resulting from high-energy trauma and aging, the development of innovative therapeutic strategies to improve bone repair based on the combination of skeletal/mesenchymal stem/stromal cells and biomimetic biomaterials is urgently needed. To this end, the use of reliable animal models is fundamental to better understanding the key cellular and molecular mechanisms that determine the healing outcomes. Of all the models, the mouse is the preferred research model because it offers a wide variety of transgenic strains and reagents for experimental analysis. However, the establishment of fractures in mice may be technically challenging due to their small size. Therefore, this article aims to demonstrate the procedures for the surgical establishment of a diaphyseal femur fracture in mice, which is stabilized with an intramedullary wire and resembles the most common bone repair process, through cartilaginous callus formation.

This protocol describes a surgical procedure for the establishment of a diaphyseal fracture in the femur of mice, which is stabilized with an intramedullary wire, for fracture healing studies.

Farelerde Diafiz Femur Kırığı Modelinin Kurulması

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

<meta charset="utf8"></head><body>Farelerde Kemik Onarımı için Standardize Mandibular Kırık Protokolü

Less-Invasive Technique for Non-stabilized Mandibular Fracture in Mouse Models

Non-stabilized fractures can be made at mandibular sites in mice, thus making it possible to analyze bone repair using an endochondral ossification mode. To most accurately reflect this process in vivo, it is necessary to have a standardized protocol to avoid excessive bone loss and soft tissue damage, particularly at the mandibular site, an anatomical site characterized by minimal access. To our knowledge, we describe for the first time a less-invasive protocol of non-stabilized mandibular fracture in mice. Adult mice are anesthetized with isoflurane and receive a preoperative dose of buprenorphine by subcutaneous injection. A submandibular approach is performed, with a skin incision made along the inferior border of the mandible. The masseter muscle is elevated in a subperiosteal plane along the mandibular ramus with a periosteal elevator. A vertical and complete fracture of the ramus is performed from the basilar border to the coronoid notch (between condylar and coronoid processes), using a piezoelectric bone surgery device under saline serum irrigation at the basilar edge and scissors to complete the fracture of the mandibular ramus. The skin approach is closed by silk sutures. This detailed procedure for non-stabilized mandibular fractures offers an efficient procedure allowing minimal bone loss and soft tissue damage. This technique ensures successful and consistent results to accurately reflect the process of endochondral bone repair in mouse models.

<meta charset="utf8"></head><body>Fare Modellerinde Stabilize Olmayan Mandibular Kırık için Daha Az İnvaziv Teknik

This protocol presents a comprehensive and efficient method for analyzing mandibular bone repair. We describe a reproducible technique for non-stabilized mandibular fracture in a mouse model, allowing analysis of the process of endochondral bone repair with minimal tissue damage and bone loss.

Fare Modellerinde Stabilize Olmayan Mandibular Kırık için Daha Az İnvaziv Teknik

Non-stabilized fractures can be made at mandibular sites in mice, thus making it possible to analyze bone repair using an endochondral ossification mode. ...