Name: murine ileocolic bowel resection with primary anastomosis
Uploaded: 2014-10-29T14:00:00.000+00:00
Duration: 8 min 49 s
Description: Watch this Scientific Journal Video about Murine Ileocolic Bowel Resection with Primary Anastomosis at JoVE.com

We would like to acknowledge the funding contributions of the Canadian Surgical Research Fund, the Edmonton Civic Employees Research Assistance Fund, and the Alberta IBD Consortium through a grant from Alberta Innovates.

les protocoles d&#39;utilisation des animaux ont été approuvés par le comité de soins et d&#39;utilisation des animaux sciences de la santé à l&#39;Université de l&#39;Alberta. 1. Préparation des instruments, les animaux et configuration opérationnelle <ol><li> Transférer les animaux à une nouvelle, propre absent de la cage de tout solide nourriture 24 heures avant l&#39;intervention. Ils peuvent avoir accès gratuitement à l&#39;eau, et le liquide alimentation ad lib jusqu&#39;à ce que le temps de la procédure. </li><li> Autoclave tous les instruments nécessaires à la procédure. Surface de travail propre et anesthésique cône de nez avec 70% d&#39;éthanol. </li><li> Mettre en place la surface d&#39;exploitation avec microscope opératoire, appareil d&#39;anesthésie et des produits d&#39;une manière qui est confortable pour le chirurgien. Les coudes du chirurgien devraient être autorisés à se reposer confortablement sur la table d&#39;opération, avec les mains et les bras dégagée par les équipements. Instruments, des sutures, des cotons-tiges, et une seringue de 10 ml doivent être placés dans une position qui permet un accès facile au cours de la procédure. </li><li> Installerlampes chauffantes puissent fournir à la fois la chaleur de l&#39;animal durant la procédure et de la lumière pour la surface d&#39;exploitation. </li><li> Remplir un tube conique de 50 ml avec une solution saline à 0,9%, et un tube de 1,5 ml avec de la vaseline et le lieu près de la surface de fonctionnement. REMARQUE: Stériliser tous les instruments et de fournitures chirurgicales. Parce que l&#39;intestin est sectionné, la procédure elle-même ne sont pas stériles. Il est considéré comme propre-contaminée. Prendre des mesures pour éviter l&#39;introduction de sources exogènes de l&#39;infection. </li></ol> 2. iléocoliques résection avec anastomose <ol><li> Induire une anesthésie par administration de 4% d&#39;isoflurane avec un débit d&#39;oxygène de 2 L / min par l&#39;intermédiaire de la coiffe jusqu&#39;à ce que le vaporisateur isoflurane fréquence respiratoire des animaux ralentit à environ 30 à 40 respirations / min. Appliquer une pression modérée sur le pied arrière de la souris pour assurer qu&#39;il n&#39;y a pas de réponse à la douleur avant de lancer la procédure. À ce stade, baissez isoflurane à 2% et le flux d&#39;oxygène à 0,5 L / min. Intermittentevérifier la réponse de la douleur au cours de la procédure et régler le débit de l&#39;isoflurane en conséquence. </li><li> Appliquez de la vaseline pour les yeux pour éviter le dessèchement pendant la chirurgie, et immobiliser la souris dans la position couchée sur le dos avec les membres sécurisées à l&#39;aide du ruban adhésif transparent. </li><li> Nettoyez l&#39;abdomen avec une solution povodine / d&#39;iode, et se transformer en nouveaux gants stériles. </li><li> Faire un 1,5 cm incision de la peau sur la ligne médiane supérieure de l&#39;abdomen à l&#39;aide de ciseaux pointu dissection pour exposer le fascia et du péritoine. Ouvrez la couche aponévrotique / péritonéale de la même façon à travers la ligne blanche pour exposer le contenu du péritoine. </li><li> En revanche chez l&#39;homme, le caecum de souris se trouve généralement dans le quadrant supérieur gauche de l&#39;abdomen. Une fois identifié, saisir doucement le caecum avec une pince et de le livrer à travers l&#39;incision. Utilisez des cotons-tiges humectés de ventilateur sur environ 3 cm de l&#39;iléon terminal extension du caecum sur une gaze stérile drapée sur la surface abdominale (figure 1A).Assurez-vous de l&#39;intestin exposé est maintenu humide avec 0,9% de solution saline pendant toute la durée de la procédure. </li><li> Identifier l&#39;artère iléo-caecale bifurcation de l&#39;artère mésentérique supérieure le long du côlon (Figure 1A) à l&#39;aide du microscope opératoire. Disséquer les tissus adjacents avasculaire de l&#39;artère iléo-colique, encercler et ligaturer l&#39;artère avec une cravate de soie 5-0. Ensuite, localisez l&#39;approvisionnement en sang régional de l&#39;iléon terminal et choisir un point de sectionnement 1,5-2 cm en amont de la jonction iléo-colique. Ligaturer les branches de cette section de l&#39;iléon comme ci-dessus. Diviser les artères avec micro dissection ciseaux. </li><li> Diviser les parties ischémiques de l&#39;iléon et du colon assurer qu&#39;il ya un apport sanguin adéquat aux extrémités sectionnées (figure 1B). Il est souvent utile à la spatule en divisant l&#39;iléon à un angle de 30 degrés pour augmenter le diamètre de la lumière de sorte qu&#39;elle corresponde plus étroitement le côlon. Une fois la partie iléo-colique intestinale a été supprimé, aligner leextrémités sectionnées de l&#39;iléon et du colon sur la toile, assurant les frontières de chaque mésentériques sont alignés. </li><li> Construire l&#39;anastomose en rapprochant l&#39;extrémité sectionnée de l&#39;iléon à l&#39;extrémité sectionnée du côlon à l&#39;aide interrompu 8-0 sutures en polypropylene (figure 1c). Le premier point est placé à la frontière mésentérique, avec des sutures ultérieures placés tous les 0,5 mm jusqu&#39;à l&#39;anastomose iléo-colique est étanche. Lors du passage de l&#39;aiguille de suture à travers l&#39;iléon et du côlon, de sorte que l&#39;arête de coupe ne sont pas aplatis, et les piqûres d&#39;aiguille sont de 0,5 mm à partir des bords de l&#39;intestin de coupe. Une anastomose typique, il faudra 14 à 16 sutures interrompues. Testez l&#39;intégrité et la perméabilité de l&#39;anastomose à la fin en faisant rouler une proximale coton-tige à distal sur l&#39;iléon de forcer contenu à travers l&#39;anastomose. Contenu de l&#39;intestin grêle devraient passer librement dans le côlon sans fuite anastomotique. </li><li> Rincer l&#39;intestin exposé avec 3-4 ml d&#39;une solution saline à 0,9% à partir de la seringue de 10 mlpour laver les selles de la surface de l&#39;intestin, et fournir l&#39;intestin dans la cavité péritonéale. En utilisant 2 ml de solution saline 0,9% rincer la cavité péritonéale, et puis les égoutter ce fluide en appliquant une légère pression à la paroi abdominale latérale. </li><li> Fermer l&#39;incision avec une suture 3-0 fonctionnement de la soie, et interrompre le flux de l&#39;isoflurane. Administrer 0,1 mg / kg de l&#39;opiacé à action prolongée -buprenorphine- voie sous-cutanée pour le contrôle de la douleur post-opératoire. </li><li> Observer les animaux sous la lampe de chaleur jusqu&#39;à ce qu&#39;ils soient mobiles puis de les transférer dans une cage sans cesse réchauffé. </li></ol> 3. Soins post-opératoires et de surveillance <ol><li> De surveiller les animaux dans une cage sans cesse réchauffé les signes de détresse pour le reste de la journée. Transférer les animaux vers le centre de protection des animaux dans une nouvelle cage stérile avec accès à l&#39;alimentation liquide et de l&#39;eau ad lib. Les animaux peuvent être hébergés en groupes de 3-4. </li><li> Effectuer un contrôle sur les animaux post-opératoires, le lendemain matin,assurer que les animaux ne semblent pas en détresse. Nourrir seul régime liquide. Si elles apparaissent mal à l&#39;aise (ie, une posture voûtée ou activité minimale) administrer une dose supplémentaire de buprénorphine sous-cutanée. Arrivée sur les animaux une fois de plus dans l&#39;après-midi le jour post-opératoire 1. </li><li> Le matin du jour post-opératoire 2, les animaux doivent apparaître complètement récupéré. Preuve de la consommation alimentaire et des selles sont des signes positifs de reprise. Maintenant, reprendre une alimentation chow solide pour les animaux. REMARQUE: Les signes de détresse comprennent une posture voûtée, toilettage pauvres, et une activité minimale. Si des signes de détresse sont au premier plan les animaux doivent être euthanasiés. </li><li> Euthanasier les animaux en induisant anesthésie profonde avec 4% d&#39;isoflurane à un débit O 2 de 2 L / min jusqu&#39;à ce que les animaux ne réagissent pas à la pression du pied. Effectuer dislocation cervicale, et d&#39;observer les signes de l&#39;euthanasie approprié. </li><li> Lignes directrices varient, se réfèrent à des institutions recommandations concernant les indicationspour et des méthodes appropriées d&#39;euthanasie chez les souris. </li></ol>

<ol>
	<li>Hallet, J., Zih, F. S., Lemke, M., Milot, L., Smith, A. J., Wong, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neo-adjuvant+chemoradiotherapy+and+multivisceral+resection+to+optimize+R0+resection+of+locally+recurrent+adherent+colon+cancer.">Neo-adjuvant chemoradiotherapy and multivisceral resection to optimize R0 resection of locally recurrent adherent colon cancer.</a> European Journal of Surgical Oncology. 40 (6), (2014).</li><li>Rutgeerts, P., Geboes, K., Vantrappen, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+history+of+recurrent+Crohn&#39;s+disease+at+the+ileocolonic+anastomosis+after+curative+surgery.">Natural history of recurrent Crohn&#39;s disease at the ileocolonic anastomosis after curative surgery.</a> Gut. 25 (6), 665-672 (1984).</li><li>Davis, D., Rivadeneira, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+colorectal+anastomoses:+leaks,+strictures,+and+bleeding.">Complications of colorectal anastomoses: leaks, strictures, and bleeding.</a> The Surgical clinics of North America. 93 (1), 61-87 (2013).</li><li>Helmrath, M. A., VanderKolk, W. E., Can, G., Erwin, C. R., Warner, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+adaptation+following+massive+small+bowel+resection+in+the+mouse.">Intestinal adaptation following massive small bowel resection in the mouse.</a> Journal of the American College of Surgeons. 183 (5), 441-449 (1996).</li><li>Dekaney, C. M., Fong, J. J., Rigby, R. J., Lund, P. K., Henning, S. J., Helmrath, 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=Expansion+of+intestinal+stem+cells+associated+with+long-term+adaptation+following+ileocecal+resection+in+mice.+American+journal+of+physiology.">Expansion of intestinal stem cells associated with long-term adaptation following ileocecal resection in mice. American journal of physiology.</a> Gastrointestinal and liver physiology. 293 (5), (2007).</li><li>Speck, K. E., De Cruz, 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=Inflammation+enhances+resection-induced+intestinal+adaptive+growth+in+IL-10+null+mice.">Inflammation enhances resection-induced intestinal adaptive growth in IL-10 null mice.</a> The Journal of surgical research. 168 (1), 62-69 (2011).</li><li>Rigby, R. J., Hunt, M. 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=A+new+animal+model+of+postsurgical+bowel+inflammation+and+fibrosis:+the+effect+of+commensal+microflora.">A new animal model of postsurgical bowel inflammation and fibrosis: the effect of commensal microflora.</a> Gut. 58 (8), 1104-1112 (2009).</li><li>Kostic, A. D., Xavier, R. J., Gevers, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Microbiome+in+Inflammatory+Bowel+Disease:+Current+Status+and+the+Future+Ahead.">The Microbiome in Inflammatory Bowel Disease: Current Status and the Future Ahead.</a> Gastroenterology. , 1-11 (2014).</li><li>Gevers, D., Kugathasan, 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=The+Treatment-Naive+Microbiome+in+New-Onset+Crohn&#39;s+Disease.">The Treatment-Naive Microbiome in New-Onset Crohn&#39;s Disease.</a> Cell host &amp; microbe. 15 (3), 382-392 (2014).</li><li>Ahmed, T., Rieder, F., Fiocchi, C., Achkar, 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=Pathogenesis+of+postoperative+recurrence+in+Crohn&#39;s+disease.">Pathogenesis of postoperative recurrence in Crohn&#39;s disease.</a> Gut. 60 (4), 553-562 (2011).</li><li>Borowiec, A., Sydora, B., 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+bowel+fibrosis+and+systemic+inflammatory+response+after+ileocolonic+anastomosis+in+IL-10+null+mice.">Small bowel fibrosis and systemic inflammatory response after ileocolonic anastomosis in IL-10 null mice.</a> Journal of Surgical Research. 178 (1), 147-154 (2012).</li><li>Devine, A. A., Gonzalez, A., 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> Impact of Ileocecal Resection and Concomitant Antibiotics on the Microbiome of the Murine Jejunum and Colon. 8 (8), (2013).</li><li>Bij, G. J., Oosterling, S. J., Beelen, R. H. J., Meijer, S., Coffey, J. C., van Egmond, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perioperative+period+is+an+underutilized+window+of+therapeutic+opportunity+in+patients+with+colorectal+cancer.">The perioperative period is an underutilized window of therapeutic opportunity in patients with colorectal cancer.</a> Annals of surgery. 249 (5), (2009).</li><li>Scott, A. D., Uff, C., Phillips, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+macrophage+function+by+suture+materials+and+anastomotic+recurrence+of+Crohn&#39;s+disease.">Suppression of macrophage function by suture materials and anastomotic recurrence of Crohn&#39;s disease.</a> The British journal of surgery. 80 (3), (1993).</li><li>Kiernan, 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=Intestinal+anastomosis+in+the+rat+facilitated+by+a+rapidly+digested+internal+splint+and+indigestible+but+absorbable+sutures.">Intestinal anastomosis in the rat facilitated by a rapidly digested internal splint and indigestible but absorbable sutures.</a> Journal of Surgical Research. 45, 427-431 (1988).</li><li>Andersen, T. 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=Action+of+matrix+metalloproteinases+at+restricted+sites+in+colon+anastomosis+repair:+an+immunohistochemical+and+biochemical+study.">Action of matrix metalloproteinases at restricted sites in colon anastomosis repair: an immunohistochemical and biochemical study.</a> Surgery. 140 (1), 72-82 (2006).</li><li>Thompson, S. K., Chang, E. Y., Jobe, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+review:+Healing+in+gastrointestinal+anastomoses,+Part+I.">Clinical review: Healing in gastrointestinal anastomoses, Part I.</a> Microsurgery. 26 (3), 131-136 (2006).</li></ol>

The authors have nothing to disclose

La souris ICR est un modèle puissant qui peut être utilisé pour étudier les effets de la chirurgie dans les maladies de l&#39;intestin. Cet article décrit un procédé de réalisation IC chez la souris avec un taux de réussite de 95% et pas de problèmes avec un retard de croissance, comme en témoigne poids stables jusqu&#39;à 28 jours après l&#39;intervention. Les défis les plus importants de la réussite ICR notamment d&#39;éviter obstructions intestinales à l&#39;anastomose et les fuites anastomotiques. <...

Ileocolic resection (ICR) is a common procedure performed in both emergent and elective situations for a variety of illnesses. Crohn&#8217;s disease and colon cancers are the two most common indications for ICR. In both illnesses, recurrence in the bowel at the site of surgery represents a major clinical problem. Local recurrence rates for colon cancer remain an issue even with the most aggressive resections1. Following ICR in Crohn&#8217;s disease, the disease most frequently (in up to 80%) recurs in the neo-terminal ileum at 1 year after surgery2. Given the impact of these two illnesses and their recurrence after surgery, it is important to understand local intestinal factors after ICR, which may have intrinsic influences on the natural history of these diseases. Further, it is also important to consider anastomotic healing after ICR. In the early post-operative period, anastomotic leaks can have devastating consequences for patients resulting in repeat surgeries, stoma creations, significant morbidity, and even mortality3. Despite the importance of this topic, our current understanding of anastomotic healing remains in its infancy as a subject of research. Animal models of ICR, in particular mice, are an excellent platform for studying the intestinal and anastomotic healing following surgery.
A mouse model of ICR was initially developed by Helmrath et al. to be used as a model of short gut syndrome4. The authors experimented with various diet regiments and suture sizes to optimize animal survival following ICR. They concluded that feeding with liquid diet in the perioperative period and using 9-0 monofilament sutures resulted in an optimal post-operative survival of 88%. Since this initial publication, ICR in mice removing 50% of the small bowel has been used in several studies to explore the dynamics of massive small bowel resection and the adaptive growth response in attempt to develop new therapies for short gut syndrome5,6.
The first application of the ICR mouse model to Crohn&#8217;s disease used the IL-10-/- mouse model, which spontaneously develops colitis7. The authors found that after ICR these animals developed inflammation in the neo-terminal ileum similar to that seen in post-operative Crohn&#8217;s disease patients, and that this inflammation was dependent on the presence of bacteria7. More recently, this model was used to explore bacterial changes induced by ICR. In Crohn&#8217;s disease there is an associated dysbiosis with relative decreases in bacteria known to have anti-inflammatory properties and increases in invasive species of bacteria8,9. The association holds true in cases of post-operative recurrence10. Two studies sought to identify microbial changes resulting from ICR. The first used IL-10 null mice, and performed denaturing gel electrophoresis to compare bacterial similarity between the small bowel and colon after ICR11. This study demonstrated that bacterial populations became similar in the small intestine and colon following ICR. A subsequent study used wild type mice and 16s pyrosequencing for phylogenetic classification of bacterial species post-operatively. This study demonstrated a marked shift in bacterial species resulting from surgery alone with Clostridium species becoming dominant as well as an increase in &#978;-proteobacteria. The results also confirmed the findings of the previous study with similar populations found in the small bowel and colon after ICR12. 
ICR is a common procedure for patients with colon cancer involving the cecum and ascending colon, and it is becoming increasingly recognized that the host-response to surgery likely contributes to both local and distant tumor recurrence13. Despite this observation, models of ICR have not been utilized for the study of colorectal cancer and post-operative recurrence. Understanding both the systemic and local immunologic changes resulting from ICR will be important in investigating future therapies. Potential pathways involved in cancer recurrence post ICR include up regulation of growth factors, which may rescue cells from apoptosis and stimulate proliferation, mechanical tumor disruption with cell shedding, and loss of immune surveillance through post-operative immunosuppression13,14. 
Mouse models of ICR have the potential to be a powerful tool for the investigation of short bowel syndrome, Crohn&#8217;s disease, and colon cancer. They may also provide lessons on how to prevent early post-operative anastomotic complications by further defining the cellular and biochemical pathways involved in healing the newly constructed anastomosis. A major barrier in utilizing the murine ICR model is the technical difficulty. The intestinal anastomosis requires the use of 8-0 or 9-0 suture, an operating microscope, and training in microsurgical techniques. The goal of this article is to provide clear instructions on how to perform ICR in mice with the goal of utilizing this procedure in models of disease.

<table><tbody><tr><td>LD101 liquid rodent diet</td><td>testdiet.com</td><td></td><td></td></tr><tr><td>0.9% NaCl</td><td>Baxter</td><td>FKE1324</td><td>Injection quality saline</td></tr><tr><td>Operating Microscope</td><td>Ziess</td><td></td><td></td></tr><tr><td>Isoflurane Anesthetic Vaporizer</td><td>Harvard Apparatus</td><td>34-0483</td><td></td></tr><tr><td>Isoflurane</td><td>Abbott&amp;nbsp;</td><td>05260-05</td><td></td></tr><tr><td>Glass plate</td><td></td><td></td><td>For operating surface</td></tr><tr><td>Cotton swabs</td><td></td><td></td><td></td></tr><tr><td>Micro Castroviejo Needle holder, curved</td><td>World Precision Instruments</td><td>503377</td><td></td></tr><tr><td>Castroviejo straight scissors</td><td>World Precision Instruments</td><td>555530S</td><td></td></tr><tr><td>Dissecting Scissors</td><td>World Precision Instruments</td><td>15922</td><td></td></tr><tr><td>Dressing Forceps x 2</td><td>World Precision Instruments</td><td>500363</td><td></td></tr><tr><td>5-0 silk pre-cut sutures</td><td>Ethicon</td><td>A182H</td><td>For vessel ligation</td></tr><tr><td>8-0 Prolene on BV130-5 needle</td><td>Ethicon</td><td>8732H</td><td>For anastomosis</td></tr><tr><td>3-0 Silk on FS-2 needls</td><td>Ethicon</td><td>8665G</td><td>For abdominal wall closure</td></tr><tr><td>Petroleum Jelly</td><td>Vaseline</td><td></td><td></td></tr><tr><td>10 ml syringe</td><td>BD biosciences</td><td></td><td></td></tr><tr><td>Povidone-iodine 7.5% surgical Scrub</td><td>&lt;a href=&quot;http://betadine.com&quot;&gt;betadine.com&lt;/a&gt;</td><td></td><td></td></tr><tr><td>Heat lamps</td><td></td><td></td><td></td></tr><tr><td>buprenorphine 0.3 mg/ml</td><td>Reckitt Benckiser Healthcare Ltd.&amp;nbsp;</td><td>PL36699/0006</td><td></td></tr></tbody></table>

murine ileocolic bowel resection with primary anastomosis

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer being two common examples. Despite the frequency of these procedures, a significant knowledge gap remains in describing the inherent effects of intestinal resection on host physiology and disease pathophysiology. This article provides detailed instructions for an ileocolic resection with primary end-to-end anastomosis in mice, as well as essential aspects of peri-operative care to maximize post-operative success. When followed closely, this procedure yields a 95% long-term survival rate, no failure to thrive, and minimizes post-operative complications of bowel obstruction and anastomotic leak. The technical challenges of performing the procedure in mice are a barrier to its wide spread use in research. The skills described in this article can be acquired without previous surgical experience. Once mastered, the murine ileocolic resection procedure will provide a reproducible tool for studying the effects of intestinal resection in models of human disease.

Les taux de mortalité et le changement de poids post-op. Les taux de mortalité suivants IC chez les souris de type sauvage 129S1 sont généralement ~ 5%. La cause la plus fréquente de la morale est une occlusion intestinale à l&#39;anastomose. Les autres causes de mortalité incluent fuite anastomotique et hernie interne entraînant une occlusion intestinale. La perte de poids peut être vu jusqu&#39;à 14 jours après l&#39;opération, mais est généralement non signif...

Watch this Scientific Journal Video about Murine Ileocolic Bowel Resection with Primary Anastomosis at JoVE.com

Murine Ileocolic Bowel Resection with Primary Anastomosis

Ileocolic resection is commonly performed in several human diseases; however, little is known regarding the impact of intestinal resection on surgical illnesses. This article provides instruction on executing the procedure in mice with high success, providing a means to study the effects of ileocolic resection in models of disease.

Murin iléocoliques résection du côlon avec anastomose primaire

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer ...

murine-ileocolic-bowel-resection-with-primary-anastomosis

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16.3K vues.  University of Alberta. Ileocolic resection is commonly performed in several human diseases; however, little is known regarding the impact of intestinal resection on surgical illnesses. This article provides instruction on executing the procedure in mice with high success, providing a means to study the effects of ileocolic resection in models of disease.

Vidéo: Murine Ileocolic Bowel Resection with Primary Anastomosis

Small Bowel Transplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis.  In this article, we provide details of the technique to supplement the video.

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

Transplantation de l&#39;intestin chez les souris

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

Recherche

Biologie

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer that often results in severe short bowel syndrome-like adverse effects including bile acid diarrhea, dehydration, electrolyte disturbances, and nutrient malabsorption. Here we introduce a murine ileal resection model, termed ileectomy, to evaluate tissue communication and the maintenance of systemic homeostasis. After ileal resection, circulating blood is permanently devoid of the ileum-specific endocrine hormone fibroblast growth factor 15 (FGF15), which releases its endocrinal inhibition of bile acid synthesis in the liver. In combination with the increased production and abolished reabsorption of bile acids after removing the ileum, mice that underwent surgery suffer from bile salt overaccumulation in the intestine and associated diarrhea, morbidity, and mortality. Novel usage of the surgery model introduced in this study may provide mechanistic and functional insights into ileal control of systemic metabolic regulation in physiology and disease.

Small intestine-dependent bile acid reabsorption and feedback inhibition of hepatic bile acid synthesis is important for systemic homeostasis and health. In this study, we describe a mouse model for ileal resection to evaluate ileectomy-induced bile malabsorption, overaccumulation, and toxicity in mouse intestine.

Suraccumulation induite par le Ileectomy de Bile dans l’intestin de souris

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

Médecine

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

Murine colostomie distale, un nouveau modèle de colite détournement chez les souris C57BL/6

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

Creation of Colonic Anastomosis in Mice

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of colonic surgeries and can occur in up to 26% of surgical anastomoses, with mortality being up to 39% for patients with such a leak. Currently, there remains a paucity of data detailing the cellular mechanisms of anastomotic healing. Devising preventative strategies and treatment modalities for anastomotic leak could be greatly potentiated by a better understanding of appropriate anastomotic healing.
A murine model is ideal as previous studies have shown that the murine anastomosis is the most clinically similar to the human case as compared with other animal models. We offer an easily reproducible murine model of colonic anastomosis in mice that will allow for further illustration of anastomotic healing.

Anastomotic leak or breakdown after surgery is a major cause of postoperative morbidity and mortality. Our surgery for creating a colonic anastomosis is a reliable and reproducible method for studying the healing mechanisms of anastomoses.

Création de l’anastomose du côlon chez la souris

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of ...

Robotic Radical Cystectomy, Pelvic Lymph Node Dissection, and Intracorporeal Ileal Conduit Urinary Diversion

The robotic approach to radical cystectomy is compelling because of its oncologic equivalence to open radical cystectomy (ORC), its association with lower surgical blood loss, its potential association with shorter hospital stay after surgery. These factors suggest that the robotic approach to radical cystectomy may be an important component of enhanced recovery programs aimed at reducing surgical morbidity. This paper describes the importance of the cranial placement of robotic trocars, the use of Cadiere forceps for atraumatic bowel grasping, pelvic lymph node dissection (PLND), and utero-enteric anastomoses. Also discussed are steps that are critical for the successful outcome of RARC. In spite of the increased operating times and associated costs and the costs of robotic surgical platforms and equipment, adoption of the robotic technique by bladder cancer surgeons has increased. This paper describes a systematic and reproducible method that details robotic extended pelvic lymph node dissection, cystectomy/cystoprostatectomy, and intracorporeal ileal conduit urinary diversion.

This paper describes robotic radical cystectomy, pelvic lymph node dissection, and intracorporeal ileal conduit urinary diversion.

The robotic approach to radical cystectomy is compelling because of its oncologic equivalence to open radical cystectomy (ORC), its association with lower ...

<meta charset="utf8"></head><body>Modèle murin pour l’étude de l’iléus postopératoire par manipulation intestinale

Postoperative Ileus Murine Model

Most patients experience postoperative ileus (POI) after surgery, which is associated with increased morbidity, mortality, and hospitalization time. POI is a consequence of mechanical damage during surgery, resulting in disruption of motility in the gastrointestinal tract. The mechanisms of POI are related to aberrant neuronal sensitivity, impaired epithelial barrier function, and increased local inflammation. However, the details remain enigmatic. Therefore, experimental murine models are crucial for elucidating the pathophysiology and mechanism of POI injury and for the development of novel therapies.
Here, we introduce a murine model of POI generated via intestinal manipulation (IM) that is similar to clinical surgery; this is achieved by mechanical damage to the small intestine by massaging the abdomen 1-3 times with a cotton swab. IM delayed gastrointestinal transit 24 h after surgery, as assessed by FITC-dextran gavage and fluorescence detection of the segmental digestive tract. Moreover, tissue swelling of the submucosa and immune cell infiltration were investigated by hematoxylin and eosin staining and flow cytometry. Proper pressure of the IM and a hyperemic effect on the intestine are critical for the procedure. This murine model of POI can be utilized to study the mechanisms of intestinal damage and recovery after abdominal surgery.

<meta charset="utf8"></head><body>Modèle postopératoire d’iléus murin

We describe a murine model of postoperative ileus generated via intestinal manipulation. Gastrointestinal transit function, pathologic changes, and immune cell activation were assessed 24 h after surgery.

Modèle postopératoire d’iléus murin

Most patients experience postoperative ileus (POI) after surgery, which is associated with increased morbidity, mortality, and hospitalization time. POI ...

Immunologie et infection

Robot-assisted Total Mesorectal Excision and Lateral Pelvic Lymph Node Dissection for Locally Advanced Middle-low Rectal Cancer

Since their approval for clinical use, da Vinci surgical robots have shown great advantages in gastrointestinal surgical operations, especially in complex procedures. The high-quality 3-D visual, multijoint arm and natural tremor filtration allow the surgeon to expose and dissect more accurately with minimal invasion. Total mesorectal excision is the standard surgical technique for the treatment of resectable rectal cancer. To reduce the lateral recurrence rate, lateral pelvic lymph node dissection can be performed, as it is a safe and feasible procedure for locally advanced middle-low rectal cancer with a high possibility of metastasis to the lateral lymph nodes. However, the complexity of the anatomic structures and the high postoperative complication rate limit its application. Recently, several surgeons have increasingly used robotic techniques for total mesorectal excision and lateral pelvic lymph node dissection. Compared with open and laparoscopic surgery, the robotic technique has several advantages, such as less blood loss, fewer blood transfusions, minimal trauma, shorter postoperative hospitalization, and quicker recovery. A robotic approach is generally regarded as a reasonable alternative for complicated procedures such as lateral pelvic lymph node dissection, although there are a limited number of high-quality prospective randomized controlled studies reporting direct evidence. Here, we provide the detailed steps of robot-assisted total mesorectal excision and lateral pelvic lymph node dissection performed at the First Affiliated Hospital of Xi&#39;an Jiaotong University.

The robotic technique described herein aims to detail a stepwise approach to robot-assisted total mesorectal excision and lateral pelvic lymph node dissection for locally advanced (T3/T4) rectal cancer located below the peritoneal reflection.

Excision mésorectale totale assistée par robot et curage ganglionnaire pelvien latéral pour le cancer du rectum localement avancé de moyenne à basse température

Since their approval for clinical use, da Vinci surgical robots have shown great advantages in gastrointestinal surgical operations, especially in complex ...

Single-Anastomosis Duodeno-Ileal Bypass with Sleeve Gastrectomy Model in Mice

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis, cardiovascular events, and cancers) through restrictive and malabsorptive mechanisms. Understanding the mechanisms by which these procedures allow such improvements often require their transposition into animals, especially in mice, because of the ease of generating genetically modified animals. Recently, the single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) has emerged as a procedure that uses both restrictive and malabsorptive effects, which is being used as an alternative to gastric bypass in case of major obesity. Thus far, this procedure has been associated with strong metabolic improvements, which has led to a marked increase in its use in daily clinical practice. However, the mechanisms underlying these metabolic effects have been poorly studied as a result of a lack of animal models. In this article, we present a reliable and reproducible model of SADI-S in mice, with a special focus on perioperative management. The description and use of this new rodent model will be helpful for the scientific community to better understand the molecular, metabolic, and structural changes induced by the SADI-S and to better define the surgical indications for clinical practice.

Single-anastomosis duodeno-ileal bypass (SADI-S) is an emerging bariatric procedure with important metabolic effects. In this article, we present a reliable and reproducible model of SADI-S in mice.

Dérivation duodéno-iléale à anastomose unique avec modèle de gastrectomie à manchon chez la souris

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes ...

Murin iléocoliques résection du côlon avec anastomose primaire

Résumé

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Chapitres dans cette vidéo

Transplantation de l'intestin chez les souris

Suraccumulation induite par le Ileectomy de Bile dans l’intestin de souris

Murine colostomie distale, un nouveau modèle de colite détournement chez les souris C57BL/6

Création de l’anastomose du côlon chez la souris

Robotic Radical Cystectomy, Pelvic Lymph Node Dissection, and Intracorporeal Ileal Conduit Urinary Diversion

Modèle postopératoire d’iléus murin

Modèle postopératoire d’iléus murin

Modèle postopératoire d’iléus murin

Excision mésorectale totale assistée par robot et curage ganglionnaire pelvien latéral pour le cancer du rectum localement avancé de moyenne à basse température

Dérivation duodéno-iléale à anastomose unique avec modèle de gastrectomie à manchon chez la souris