Name: a rabbit aortic valve stenosis model induced by direct balloon injury
Uploaded: 2023-03-31T14:40:00
Description: Watch this Scientific Journal Video about A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury at JoVE.com

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MSIT) (No. 2020R1A4A3079570), the Ministry of Education (No. 2021R1I1A1A01051425), and the Industrial Strategic Technology Development Program (No. 20014873) funded by the Ministry of Trade, Industry &#38; Energy, Republic of Korea.

All animal research procedures were approved and performed in accordance with the Laboratory Animals Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Animal Experiments provided by the Institutional Animal Care and Use Committee (IACUC) at the College of Medicine of The Catholic University of Korea (approval number: CUMC-2021-0176-05). The present study utilized 3 month old male New Zealand white (NZW) rabbits weighing 3.5-4.0 kg, which were maintained under standard ...

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Animal AVS models are commonly used to study the pathological aspects of AVS, including the initiation and progression of AVS. This protocol introduces a new rabbit AVS model induced by a direct balloon injury to the aortic valve. In this study, the aortic valve injury model showed significant leaflet thickening and calcification. Compared to the mild AVS model induced by dietary supplementation, the aortic valve in the direct balloon injury model was selectively injured, leading to thickened cusps and restricted motion,...

It is increasingly recognized that the use of appropriate animal models can contribute to a better understanding of the pathological mechanisms underlying aortic valve stenosis (AVS) due to the lack of access to reliable sources of diseased human aortic valves associated with the progression of aortic stenosis (AS). Among the various animal models for studying AVS, rabbits are one of the most commonly used large-animal AVS models, and the AVS rabbit model is induced either through cholesterol/vitamin D2 supplementation or genetic manipulation1,2,3,4.
Although rabbit AVS models have provided significant insight into the development and progression of AVS, it still remains challenging to induce AVS consistently and reproducibly, as seen in our preliminary experiments.
In addition to diet-induced and genetically susceptible animal models, a new model of AVS has been established through direct mechanical injury in mice5,6. The mechanical injury model successfully induces aortic stenosis and represents a simple and reproducible AVS model in wild-type mice. To the best of our knowledge, there have been no prior studies examining the effects of a mechanical injury on the aortic valve in rabbit models. Thus, this study provides a new procedure for inducing AVS in male New Zealand white rabbits through a direct balloon injury to the aortic valve, which can accurately mimic the condition of valvular aortic stenosis. This protocol includes step-by-step descriptions of the preparation, the surgical procedure, and the post-operative care, which are useful for inducing reproducible AVS rabbit models.

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a rabbit aortic valve stenosis model induced by direct balloon injury

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of access to reliable sources of diseased human aortic valves. Among the various animal models, AVS rabbit models are one of the most commonly used in large animal studies. However, traditional AVS rabbit models require a long-term period of dietary supplementation and genetic manipulation to induce significant stenosis in the aortic valve, limiting their use in experimental studies. To address these limitations, a new AVS rabbit model is proposed, in which stenosis is induced by a direct balloon injury to the aortic valve. The present protocol describes a successful technique for inducing AVS in New Zealand white (NZW) rabbits, with step-by-step procedures for the preparation, the surgical procedure, and the post-operative care. This simple and reproducible model offers a promising approach for studying the initiation and progression of AVS and provides a valuable tool for investigating the underlying pathological mechanisms of the disease.

Rabbit AVS model induced by aortic valve injury 
To induce the rabbit AVS model, male NZW rabbits weighing 3.5-4.0 kg were used for this study. According to the surgical procedures described in step 2 (Figure 2), the AVS model was established by aortic valve injury, which resulted in mechanical aortic valve degeneration and calcification. The control group included rabbits fed with a 0.5% cholesterol-enriched diet (high-cholesterol, HC) and 50,000 U of vitamin D2 (VitD2...

Watch this Scientific Journal Video about A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury at JoVE.com

A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury

An appropriate animal model is needed to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) and to evaluate the efficacy of therapeutic interventions. The present protocol describes a new procedure for developing the AVS rabbit model via a direct balloon injury in vivo.

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of ...

Annual models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis because of the lack of access to reliable sources of diseased human aortic valves. Traditional aortic valve stenosis rabbit models require long-term dietary supplementation and genetic manipulation to induce significant stenosis in the aortic valve, limiting their use in experimental studies. This technique describes a new procedure for inducing aortic valve stenosis in rabbits through a direct ballon injury to the aortic valve, which can accurately mimic the condition of a human with aortic stenosis.Demonstrating the procedure will be Eun-Hye Park, Jin-Moo Kim, and Eunmi Lee, research specialists from my regulatory. Begin preparing the dilation balloon catheter set by connecting the in deflation device filled with the one-to-one mixture of saline and commercially available contrast medium to the Luer lock part of the balloon catheter. After filling the balloon with an inflation solution, remove the air from the balloon catheter.Begin the surgery procedure by inserting a 24 gauge intravenous or IV catheter into the marginal auricular vein of the anesthetized rabbit. Connect an infusion set with 100 units per kilogram of heparin heparinized saline. Next, connect the rabbit with a multi-parameter veterinary monitor to continuously monitor the vital signs such as the oxygen saturation signal or SpO2 temperature and blood pressure.After placing the rabbit in a supine position on an operating table equipped with a C-arm fluoroscopy, remove the hair from the ventral neck area using animal hair clippers. Sterilize the incision area with iodine and cover the rabbit with surgical towels. To position the rabbit's heart, turn on the C-arm and select the fluoroscopic mode for cardiac imaging.Adjust the rabbit's position to ensure the heart is at the center of the imaging field. Once the position is adjusted, make a longitudinal incision of approximately three centimeter in the skin of the neck and use surgical scissors to cut the fascia and fat tissue. Then carefully separate the muscles until approximately 3 to 3.5 centimeters of the left common carotid artery, or LCCA, is exposed.Ligate the LCCA with a 3-0 silk suture at the top and end of the exposed LCCA to stop the blood flow. Insert a 22 gauge IV catheter into the LCCA. Then introduce a 0.016 inch guide wire into the left ventricle through the IV catheter, ensuring that the tip of the catheter is properly positioned in the imaging field of the C-arm.Next, withdraw the IV catheter, leaving the guide wire before placing a 4 French sheath over the guide wire into the LCCA to introduce the balloon catheter. Carefully insert the eight millimeter balloon catheter over the guide wire into the aortic valve under C-arm fluoroscopic guidance. Place the balloon catheter tip approximately one to two centimeters distal to the aortic valve and inflate the balloon by purging the inflation solution with a pressure inflator at six atmospheres.Next, advance the balloon into the left ventricle or LV apex and pull it back into the LV outlet. Repeat this procedure five times before deflating the balloon. Withdraw the balloon catheter and guide wire, then slowly remove the sheath from the LCCA and immediately tie the LCCA with the suture on the way down to the aortic valve.Clean the incision area with saline to remove the blood clots. Inspect the punctured site for arterial bleeding before closing the muscle and skin with a 3-0 non-absorbable suture. Once done, sterilize the wound using iodine.After the surgery, remove the monitoring patches and clips and keep the rabbit in an intensive care incubator. After eight weeks of the balloon injury, place the anesthetized rabbit on an echo table in the supine position. Apply ultrasound transducer gel to the chest after shaving the chest area.Adjust the transducer to obtain the peristernal long-axis view and the peristernal short-axis view of the aortic valve. Use mode imaging to record images of the aortic valve and save the images for later analysis. The structural change assessment in the aortic valve revealed that eight weeks after the aortic valve injury, the cusps were thickened and the motion was restricted in the injured rabbits fed with the high cholesterol and vitamin D2 diet compared to the control or wild type rabbits, and the rabbits fed the high cholesterol and vitamin D2 diet without valve injury.The rabbits were sacrificed and the hearts were excised eight weeks after the injury for histological analysis of the aortic valve. The aortic valve stained with Masson's trichrome showed increased thickness of the aortic valve cusps in the injured group compared to the control in high cholesterol and vitamin-D2-diet-induced groups. Alizarin Red staining and Von Kossa staining were performed to compare the degree of valvular calcium deposits indicated negligible calcium deposits in the valvular leaflets in the high cholesterol and vitamin-D2-diet induced group.In contrast, significant calcific deposits were observed in the balloon injured group. Successful technique for inducing arctic valve stenosis, in the rabbit model is demonstrated here with a step-by-step explanation of a direct balloon injury to the aortic valve. This simple and reproducible model offers a promising approach for studying the initiation and progression of aortic valve stenosis, and provides a valuable tool for investigating the underlying pathological mechanisms of aortic valve stenosis.

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