Name: creation of two saccular elastase-digested aneurysms with different hemodynamics in one rabbit
Uploaded: 2021-04-15T15:00:00
Description: Watch this Scientific Journal Video about Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit at JoVE.com

The authors thank Prof Hans Rudolf Widmer, Dr Luca Remonda, and Prof Javier Fandino for their scientific support and technical contribution to this work. A special thanks to Olgica Beslac for her advice during the procedures and Kay Nettelbeck for his assistance. Furthermore, we thank Daniela Casoni DVM, PhD and med. vet. Luisana Garcia, PD Dr Alessandra Bergadano, and Dr Carlotta Detotto for their dedicated veterinary support.

NOTE: The experiment was approved by the Local Committee for Animal Care of the Canton Bern, Switzerland (Application Number BE108/16), and all animal care and procedures were performed in accordance with institutional guidelines and 3R principles15,16. Data are reported according the ARRIVE guidelines. Peri-operative management was conducted by a board-certified veterinarian anesthesiologist. For the study, female New Zealand white rabbits, with a mean weight of...

<ol>
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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=Long-term+patency+of+elastase-induced+aneurysm+model+in+rabbits.">Long-term patency of elastase-induced aneurysm model in rabbits.</a> American Journal of Neuroradiology. 27 (1), 139-141 (2006).</li><li>Short, J. 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The most common technique for aneurysm creation involves the creation of a stump aneurysm at the origin of the right CCA, either through an open or an endovascular method. The model has been validated to be a stable non-growing aneurysm that remains open with time20,21. The second possible technique involves the microsurgical creation of an arterial bifurcation aneurysm by anastomosing the right CCA on the left one and suturing an aneurysm pouch on the bifurcatio...

Intracranial aneurysm is a severe condition with a mortality rate after rupture reaching 50% and long-term disability in 10-20% of patients1. The last decade has seen a rapid development of endovascular treatment options but, at the same time, also an increasing rate of recurrence with up to 33% of aneurysm recanalization after coiling2,3. To better understand the pathophysiology underlying aneurysm occlusion and recanalization, as well as for the development and testing of new endovascular devices, there is currently a need for reliable preclinical models whose hemodynamic, morphological, and histologic characteristics mimic those of human intracranial aneurysms4,5,6. As of today, there is no defined model as a standard for preclinical trial, and a large range of species and techniques are available to researchers7,8.
However, the rabbit is a species of particular interest due to the size and hemodynamic similarities between its neck arteries and the human cerebral vessels, as well as its similar coagulation and thrombolysis profiles. Several models with elastase-digested saccular aneurysms on the common carotid arteries (CCAs) have shown qualitative and quantitative similarities with human intracranial aneurysms in terms of flow conditions, geometric features, and wall characteristics9,10,11,12. This study aims to describe a technique to create a new rabbit aneurysm model with both stump and bifurcation elastase-digested aneurysms in the same animal. The surgical techniques are inspired by those of Hoh et al.13 and Wanderer et al.14 with slight modifications to provide a good standardization and reproducibility and to ensure low mortality and morbidity.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-0 resorbable suture</td>
			<td>Ethicon Inc., USA</td>
			<td>VCP292ZH</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 resorbable suture</td>
			<td>Ethicon Inc., USA</td>
			<td>VCP304H</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 non absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>C0766070</td>
			<td></td>
		</tr>
		<tr>
			<td>9-0 non absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>G1111140</td>
			<td></td>
		</tr>
		<tr>
			<td>Adrenaline</td>
			<td>Amino AG</td>
			<td>1445419</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Amiodarone</td>
			<td>Helvepharm AG</td>
			<td>5078567</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Anesthesia machine</td>
			<td>Dr&#228;ger</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Aspirin</td>
			<td>Sanofi-Aventis (Suisse) SA</td>
			<td>622693</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Atropine</td>
			<td>Labatec Pharma SA</td>
			<td>6577083</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Bandpass filter blue</td>
			<td>Thorlabs</td>
			<td>FD1B</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bandpass filter green</td>
			<td>Thorlabs</td>
			<td>FGV9</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Biemer vessel clip (2x)</td>
			<td>B. Braun Medical AG, Aesculap, Switzerland</td>
			<td>FD560R</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bispectral index (neonatal)</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Blood pressure cuff (neonatal)</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bycilces spotlight</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Clamoxyl</td>
			<td>GlaxoSmithKline AG</td>
			<td>758808</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Dexmedetomidine</td>
			<td>Ever Pharma</td>
			<td>136740-1</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Elastase</td>
			<td>Sigma Aldrich</td>
			<td>E7885</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocardiogram electrodes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ephedrine</td>
			<td>Amino AG</td>
			<td>1435734</td>
			<td></td>
		</tr>
		<tr>
			<td>Esmolol</td>
			<td>OrPha Swiss GmbH</td>
			<td>3284044</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl (intravenous use)</td>
			<td>Janssen-Cilag AG</td>
			<td>98683</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl (transdermal)</td>
			<td>Mepha Pharma AG</td>
			<td>4008286</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoresceine</td>
			<td>Curatis AG</td>
			<td>5030376</td>
			<td></td>
		</tr>
		<tr>
			<td>Fragmin</td>
			<td>Pfizer PFE Switzerland GmbH</td>
			<td>1906725</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad or heating forced-air warming system</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isotonic sodium chloride solution (0.9%)</td>
			<td>Fresenius KABI</td>
			<td>336769</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Pfizer PFE Switzerland GmbH</td>
			<td>342261</td>
			<td></td>
		</tr>
		<tr>
			<td>lid retractor</td>
			<td></td>
			<td></td>
			<td>Approach</td>
		</tr>
		<tr>
			<td>Lidocaine</td>
			<td>Streuli Pharma AG</td>
			<td>747466</td>
			<td></td>
		</tr>
		<tr>
			<td>Longuettes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Metacam</td>
			<td>Boehringer Ingelheim</td>
			<td>P7626406</td>
			<td>Medication</td>
		</tr>
		<tr>
			<td>Methadone</td>
			<td>Streuli Pharma AG</td>
			<td>1084546</td>
			<td>Sedaton</td>
		</tr>
		<tr>
			<td>Micro-forceps&#160; curved</td>
			<td>Ulrich Swiss, Switzerland</td>
			<td>U52-015-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-forceps&#160; straight 2x</td>
			<td>Ulrich Swiss, Switzerland</td>
			<td>U52-010-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscissors</td>
			<td>Ulrich Swiss , Switzerland</td>
			<td>U52-327-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Accord Healthcare AG</td>
			<td>7752484</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle 23 G</td>
			<td></td>
			<td></td>
			<td>arteriotomy</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>O2-Face mask</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Operation microscope</td>
			<td>Wild Heerbrugg</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Papaverin</td>
			<td>Bichsel</td>
			<td></td>
			<td>topical application</td>
		</tr>
		<tr>
			<td>Povidone iodine</td>
			<td>Mundipharma Medical Company</td>
			<td></td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Prilocaine-lidocaine creme</td>
			<td>Emla</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol</td>
			<td>B. Braun Medical AG, Switzerland</td>
			<td></td>
			<td>General anesthesia</td>
		</tr>
		<tr>
			<td>Pulse oxymeter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rectal temperature probe (neonatal)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer Lactate</td>
			<td>Bioren Sintetica SA</td>
			<td></td>
			<td>Infusion</td>
		</tr>
		<tr>
			<td>Ropivacain</td>
			<td>Aspen Pharma Schweiz GmbH</td>
			<td>1882249</td>
			<td>Local anesthesia</td>
		</tr>
		<tr>
			<td>Scalpell</td>
			<td>Swann-Morton</td>
			<td>210</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal shaver</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Soft tissue forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Soft tissue spreader</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel sponge bowls</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile micro swabs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stethoscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgery drape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes 1 mL, 2 mL, and 5 mL</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-Buffer</td>
			<td>Sigma Aldrich</td>
			<td>93302</td>
			<td>Elastase solution</td>
		</tr>
		<tr>
			<td>Vascular clip applicator</td>
			<td>B. Braun, Germany</td>
			<td>FT495T</td>
			<td></td>
		</tr>
		<tr>
			<td>Vein and arterial catheter 22 G</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>vessel loop</td>
			<td></td>
			<td></td>
			<td>Approach</td>
		</tr>
		<tr>
			<td>video camera or smartphone</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vitarubin</td>
			<td>Streuli Pharma AG</td>
			<td>6847559</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil titan standard clip (2x)</td>
			<td>B. Braun Medical AG, Aesculap, Switzerland</td>
			<td>FT242T</td>
			<td></td>
		</tr>
	</tbody>
</table>

creation of two saccular elastase-digested aneurysms with different hemodynamics in one rabbit

Preclinical animal models with hemodynamic, morphologic, and histologic characteristics close to human intracranial aneurysms play a key role in the understanding of the pathophysiological processes and the development and testing of new therapeutic strategies. This study aims to describe a new rabbit aneurysm model that allows the creation of two elastase-digested saccular aneurysms with different hemodynamic conditions within the same animal.
Five female New Zealand white rabbits with a mean weight of 4.0 (&#177; 0.3) kg and mean age of 25 (&#177;5) weeks underwent microsurgical stump and bifurcation aneurysm creation. One aneurysm (stump) was created by right common carotid artery (CCA) exposure at its origin at the brachiocephalic trunk. A temporary clip was applied at the CCA origin and another, 2 cm above. This segment was treated with a local injection of 100 U of elastase for 20 min. A second aneurysm (bifurcation) was created by suturing an elastase-treated arterial pouch into the end-to-side anastomosis of the right CCA to left CCA. Patency was controlled by fluorescence angiography immediately after creation.
The average duration of surgery was 221 min. The creation of two aneurysms in the same animal was successful in all rabbits without complication. All aneurysms were patent immediately after surgery except for one bifurcation aneurysm, which showed an extreme tissue reaction due to elastase incubation and an immediate intraluminal thrombosis. No mortality was observed during surgery and up to one-month follow-up. Morbidity was limited to a transient vestibular syndrome (one rabbit), which recovered spontaneously within one day.
Demonstrated here for the first time is the feasibility of creating a two-aneurysm rabbit model with stump and bifurcation hemodynamic characteristics and highly degenerated wall conditions. This model allows the study of the natural course and potential treatment strategies on the basis of aneurysm biology under different flow conditions.

The creation of a stump and a bifurcation aneurysm was successful in all 5 New Zealand white rabbits without intraoperative complications. No mortality was observed during surgery or during the follow-up period of 24 &#177; 2 days. One rabbit experienced postoperative complications with a vestibular syndrome and a blindness of the right side. The animal recovered completely and spontaneously after 24 h. This complication did not interfere with its normal activities (free movements, water and food intake, interactions wit...

Watch this Scientific Journal Video about Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit at JoVE.com

Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit

This protocol describes the steps for the creation of a rabbit model with two elastase-digested aneurysms with different hemodynamics (stump and bifurcation constellation). This allows the testing of novel endovascular devices in aneurysms with different angioarchitecture and hemodynamic conditions within a single animal.

Preclinical animal models with hemodynamic, morphologic, and histologic characteristics close to human intracranial aneurysms play a key role in the ...

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