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>
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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. 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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 &#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

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 ...

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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

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Yara Şifa murin Modeli

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Tıp

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombotic İskemi: Fare İnme Araştırmalar bir Minimal İnvaziv ve tekrarlanabilir fotokimyasal Kortikal Lezyon Model

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

Tibial Sinir transeksiyonu - Farelerde denervasyon bağlı İskelet Kas Atrofi için bir Standart Model

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

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 ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

Farelerde Minimal İnvaziv Enine Aort Konstrüksiyon

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Minimally Invasive Muscle Embedding (MIME) - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis

Skeletal muscle possesses regenerative capacity due to tissue-resident, muscle-fiber-generating (myogenic) satellite cells (SCs), which can form new muscle fibers under the right conditions. Although SCs can be harvested from muscle tissue and cultured in vitro, the resulting myoblast cells are not very effective in promoting myogenesis when transplanted into host muscle. Surgically exposing the host muscle and grafting segments of donor muscle tissue, or the isolated muscle fibers with their SCs onto host muscle, promotes better myogenesis compared to myoblast transplantation. We have developed a novel technique that we call Minimally Invasive Muscle Embedding (MIME). MIME involves passing a surgical needle through the host muscle, drawing a piece of donor muscle tissue through the needle track, and then leaving the donor tissue embedded in the host muscle so that it may act as a source of SCs for the host muscle. Here we describe in detail the steps involved in performing MIME in an immunodeficient mouse model that expresses a green fluorescent protein (GFP) in all of its cells. Immunodeficiency in the host mouse reduces the risk of immune rejection of the donor tissue, and GFP expression enables easy identification of the host muscle fibers (GFP+) and donor-cell-derived muscle fibers (GFP-). Our pilot data suggest that MIME can be used to implant an extensor digitorum longus (EDL) muscle from a donor mouse into the tibialis anterior (TA) muscle of a host mouse. Our data also suggest that when a myotoxin (barium chloride, BaCl2) is injected into the host muscle after MIME, there is evidence of donor-cell-derived myogenesis in the host muscle, with approximately 5%, 26%, 26% and 43% of the fibers in a single host TA muscle showing no host contribution, minimal host contribution, moderate host contribution, and maximal host contribution, respectively.

Minimally Invasive Muscle Embedding MIME - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis

We describe a novel experimental technique that we call Minimally Invasive Muscle Embedding (MIME), which is based on the evidence that skeletal muscle tissue contains viable myogenic cells that can facilitate donor-cell-mediated myogenesis when implanted into a host muscle.

Minimal invaziv kas (MIME) katıştırma - donör-Hücre-aracılı Myogenesis kolaylaştırmak için yeni bir deneysel teknik

Skeletal muscle possesses regenerative capacity due to tissue-resident, muscle-fiber-generating (myogenic) satellite cells (SCs), which can form new ...

Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure overload-induced left ventricular hypertrophy (LVH) and its progression to heart failure. In the majority of the reported investigations, this procedure is performed with intubation and ventilation of the animal which renders it demanding and time-consuming and adds to the surgical burden to the animal. The aim of this protocol is to describe a simplified technique of minimally invasive TAC without intubation and ventilation of mice. Critical steps of the technique are emphasized in order to achieve low mortality and high efficiency in inducing LVH.
Male C57BL/6 mice (10-week-old, 25-30 g, n=60) were anesthetized with a single intraperitoneal injection of a mixture of ketamine and xylazine. In a spontaneously breathing animal following a 3-4 mm upper partial sternotomy, a segment of 6/0 silk suture threaded through the eye of a ligation aid was passed under the aortic arch and tied over a blunted 27-gauge needle. Sham-operated animals underwent the same surgical preparation but without aortic constriction. The efficacy of the procedure in inducing LVH is attested by a significant increase in the heart/body weight ratio. This ratio is obtained at days 3, 7, 14 and 28 after surgery (n = 6 - 10 in each group and each time point). Using our technique, LVH is observed in TAC compared to sham animals from day 7 through day 28. Operative and late (over 28 days) mortalities are both very low at 1.7%.
In conclusion, our cost-effective technique of minimally invasive TAC in mice carries very low operative and post-operative mortalities and is highly efficient in inducing LVH. It simplifies the operative procedure and reduces the strain put on the animal. It can be easily performed by following the critical steps described in this protocol.

The aim of this protocol is to describe step-by-step the technique of minimally invasive transverse aortic constriction (TAC) in mice. By elimination of intubation and ventilation which are mandatory for the commonly used standard procedure, minimally invasive TAC simplifies the operative procedure and reduces the strain put on the animal.

Minimal invaziv transvers aort daralma farelerde sol ventrikül hipertrofisi indüksiyon için tekniği

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure ...

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 ...

Electromagnetic Navigation Transthoracic Nodule Localization for Minimally Invasive Thoracic Surgery

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or excision. Many of these nodules are identified and excised via minimally invasive thoracic surgery; however, subcentimeter and subsolid nodules are frequently difficult to identify intra-operatively. This can be mitigated by the use of electromagnetic transthoracic needle localization. This protocol delineates the step-by-step process of electromagnetic localization from the pre-operative period to the postoperative period and is an adaptation of the electromagnetically guided percutaneous biopsy previously described by Arias et al. Pre-operative steps include obtaining a same day CT followed by the generation of a three-dimensional virtual map of the lung. From this map, the target lesion(s) and an entry site are chosen. In the operating room, the virtual reconstruction of the lung is then calibrated with the patient and the electromagnetic navigation platform. The patient is then sedated, intubated, and placed in the lateral decubitus position. Using a sterile technique and visualization from multiple views, the needle is inserted into the chest wall at the prechosen skin entry site and driven down to the target lesion. Dye is then injected into the lesion and, then, continuously during needle withdrawal, creating a tract for visualization intra-operatively. This method has many potential benefits when compared to the CT-guided localization, including a decreased radiation exposure and decreased time between the dye injection and the surgery. Dye diffusion from the pathway occurs over time, thereby limiting intra-operative nodule identification. By decreasing the time to surgery, there is a decrease in wait time for the patient, and less time for dye diffusion to occur, resulting in an improvement in nodule localization. When compared to electromagnetic bronchoscopy, airway architecture is no longer a limitation as the target nodule is accessed via a transparenchymal approach. Details of this procedure are described in a step-by-step fashion.

Presented here is a protocol for lung nodule localization using dye marking via electromagnetically navigated transthoracic needle access. The technique described here can be accomplished in the peri-operative period to optimize nodule localization and to successful resection when performing minimally invasive thoracic surgery.

Minimal İnvaziv Göğüs Cerrahisinde Elektromanyetik Navigasyon Transtorasik Nodül Lokalizasyonu

The increased use of chest computed tomography (CT) has led to an increased detection of pulmonary nodules requiring diagnostic evaluation and/or ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Rekonstrüktif Mikrocerrahide İskemi-Reperfüzyon Hasarı Nın İnceleştirilmesinde Klinik Öncesi Sıçan Modeli

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...