Name: operating transverse aortic constriction with absorbable suture to obtain transient myocardial hypertrophy
Uploaded: 2020-09-09T14:45:00
Description: Watch this Scientific Journal Video about Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy at JoVE.com

This work was supported by grants from the National Natural Science Foundation of China (81770271; to Y, Liao), the Joint Funds of the National Natural Science Foundation of China (U1908205; to Y, Liao), and the Municipal Planning Projects of Scientific Technology of Guangzhou (201804020083; to Dr Liao).

All procedures were carried out following the guidelines of the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised in 1996). C57BL/6J male mice (8&#8211;10 weeks, 20&#8211;25 g) were provided by the Animal Center of South Medical University.
1. Preoperative preparation
<ol>
	<li>Pinch off the tip of a 25 G needle with a needle holder and blunt it with a hard object like the holder.</li>
	<li>Pass a 5&#8211;0 absor...

<ol>
	<li>Murry, C. E., Jennings, R. B., Reimer, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preconditioning+with+ischemia:+a+delay+of+lethal+cell+injury+in+ischemic+myocardium.">Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium.</a> Circulation. 74 (5), 1124-1136 (1986).</li><li>Ban, K., 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=Phosphatidylinositol+3-kinase+gamma+is+a+critical+mediator+of+myocardial+ischemic+and+adenosine-mediated+preconditioning.">Phosphatidylinositol 3-kinase gamma is a critical mediator of myocardial ischemic and adenosine-mediated preconditioning.</a> Circulation Research. 103 (6), 643-653 (2008).</li><li>Wu, Z. K., Iivainen, T., Pehkonen, E., Laurikka, J., Tarkka, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemic+preconditioning+suppresses+ventricular+tachyarrhythmias+after+myocardial+revascularization.">Ischemic preconditioning suppresses ventricular tachyarrhythmias after myocardial revascularization.</a> Circulation. 106 (24), 3091-3096 (2002).</li><li>Hausenloy, D. J., Yellon, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preconditioning+and+postconditioning:+underlying+mechanisms+and+clinical+application.">Preconditioning and postconditioning: underlying mechanisms and clinical application.</a> Atherosclerosis. 204 (2), 334-341 (2009).</li><li>Heusch, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+basis+of+cardioprotection:+signal+transduction+in+ischemic+pre-,+post-,+and+remote+conditioning.">Molecular basis of cardioprotection: signal transduction in ischemic pre-, post-, and remote conditioning.</a> Circulation Research. 116 (4), 674-699 (2015).</li><li>Lund, O., Emmertsen, K., Dorup, I., Jensen, F. T., Flo, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regression+of+left+ventricular+hypertrophy+during+10+years+after+valve+replacement+for+aortic+stenosis+is+related+to+the+preoperative+risk+profile.">Regression of left ventricular hypertrophy during 10 years after valve replacement for aortic stenosis is related to the preoperative risk profile.</a> European Heart Journal. 24 (15), 1437-1446 (2003).</li><li>Rockman, H. 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=Segregation+of+atrial-specific+and+inducible+expression+of+an+atrial+natriuretic+factor+transgene+in+an+in+vivo+murine+model+of+cardiac+hypertrophy.">Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy.</a> Proceedings of the National Academy of Sciiences of the United States of America. 88 (18), 8277-8281 (1991).</li><li>Wei, X., 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=Myocardial+hypertrophic+preconditioning+attenuates+cardiomyocyte+hypertrophy+and+slows+progression+to+heart+failure+through+upregulation+of+S100A8/A9.">Myocardial hypertrophic preconditioning attenuates cardiomyocyte hypertrophy and slows progression to heart failure through upregulation of S100A8/A9.</a> Circulation. 131 (17), 1506-1517 (2015).</li><li>Huang, J., 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=Ultrasound+biomicroscopy+validation+of+a+murine+model+of+cardiac+hypertrophic+preconditioning:+comparison+with+a+hemodynamic+assessment.">Ultrasound biomicroscopy validation of a murine model of cardiac hypertrophic preconditioning: comparison with a hemodynamic assessment.</a> American Journal of Physiology. Heart and Circulatory Physiology. 313 (1), 138-148 (2017).</li><li>Oka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac-specific+deletion+of+Gata4+reveals+its+requirement+for+hypertrophy,+compensation,+and+myocyte+viability.">Cardiac-specific deletion of Gata4 reveals its requirement for hypertrophy, compensation, and myocyte viability.</a> Circulation Research. 98 (6), 837-845 (2006).</li><li>Li, 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=Assessment+of+cardiac+morphological+and+functional+changes+in+mouse+model+of+transverse+aortic+constriction+by+echocardiographic+imaging.">Assessment of cardiac morphological and functional changes in mouse model of transverse aortic constriction by echocardiographic imaging.</a> Journal of Visualized Experiments. (112), e54101 (2016).</li><li>Veldhuizen, R. A., Slutsky, A. S., Joseph, M., McCaig, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+mechanical+ventilation+of+isolated+mouse+lungs+on+surfactant+and+inflammatory+cytokines.">Effects of mechanical ventilation of isolated mouse lungs on surfactant and inflammatory cytokines.</a> The European Respiratory Journal. 17 (3), 488-494 (2001).</li><li>Wang, Q., 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=Induction+of+right+ventricular+failure+by+pulmonary+artery+constriction+and+evaluation+of+right+ventricular+function+in+mice.">Induction of right ventricular failure by pulmonary artery constriction and evaluation of right ventricular function in mice.</a> Journal of Visualized Experiments. (147), e59431 (2019).</li><li>Eichhorn, 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=A+closed-chest+model+to+induce+transverse+aortic+constriction+in+mice.">A closed-chest model to induce transverse aortic constriction in mice.</a> Journal of Visualized Experiments. (134), e57397 (2018).</li><li>Tavakoli, R., Nemska, S., Jamshidi, P., Gassmann, M., Frossard, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+of+minimally+invasive+transverse+aortic+constriction+in+mice+for+induction+of+left+ventricular+hypertrophy.">Technique of minimally invasive transverse aortic constriction in mice for induction of left ventricular hypertrophy.</a> Journal of Visualized Experiments. (127), e56231 (2017).</li></ol>

There is still a vastly underexplored area in cardiac non-ischemic preconditioning. Based on our previous studies, we switched to using absorbable sutures to improve the myocardial hypertrophic preconditioning model.
In previous reports, many investigators used silk suture to constrict the aortic arch8,14,15. Silk suture was easily available and was often used for surgical wound suture, tissue ligatio...

Ischemic preconditioning is a phenomenon that induces brief non-lethal episodes of ischemia and reperfusion to the heart and has the capacity to dramatically reduce myocardial injury1. Given the obvious clinical implications of ischemic preconditioning, such as limiting myocardial infarct size2 and suppressing ventricular tachyarrhythmias after myocardial revascularization3, there have been lots of research to dissect the mechanisms underlying cardio-protective effects induced by preconditioning4,5. In contrast, other non-ischemic types of preconditioning have received relatively little attention. Cardiac hypertrophy may be blunted in patients with aortic stenosis undergoing aortic valve replacement6. Wherever the state of pathological myocardial hypertrophy exists, the principle of preconditioning is rarely reported.
In 1991, Rockman et al. firstly established a mouse model of left ventricular hypertrophy by transverse aortic constriction (TAC)7. By operating TAC twice in mice, we have previously proved that myocardial hypertrophic preconditioning (MHP) leads to transient hypertrophic stimulation in the heart thereby making the heart more resistant to sustained hypertrophic stress in the future8. The characteristics of the MHP model have been validated by ultrasound biomicroscope and hemodynamic assessment9. Key points in constructing the model was to perform thoracotomy three times, TAC for a week, debanding for a week, and secondary TAC for 6 weeks. However, debanding could cause bleeding, which made it difficult to be mastered by novices and difficult to be popularized. In addition, it is also a technical challenge to intubate mice. Improper intubation could cause tracheal injury, pneumothorax, and even death in mice. So, it is necessary and valuable to improve some procedures while constructing the MHP model.
To reduce the difficulty of the model and increase its success, we switched to absorbable sutures for the first TAC and monitored the model&#39;s success by measuring pressure gradient across the aortic constriction under echocardiography10. Based on our preliminary experiment, it would be difficult to induce sufficient myocardial hypertrophy in mice with too low-pressure gradient, while mice with too high-pressure gradient would develop acute heart failure or even die. The ideal pressure gradient for the model ranges from 40&#8211;80 mmHg11. In addition, this experiment did not rely on a ventilator, which could effectively avoid ventilator-associated technical manipulation and injury12.

<table>
	<tbody>
		<tr>
			<td>Absorbable suture (5-0)</td>
			<td>Shandong Kang Lida Medical Products Co., Ltd</td>
			<td>5-0</td>
			<td>Ligation</td>
		</tr>
		<tr>
			<td>Animal ultrasound system VEVO2100</td>
			<td>Visual Sonic</td>
			<td>VEVO2100</td>
			<td>Echocardiography</td>
		</tr>
		<tr>
			<td>Cold light illuminator</td>
			<td>Olympus</td>
			<td>ILD-2</td>
			<td>Light</td>
		</tr>
		<tr>
			<td>Heat pad- thermostatic surgical system (ALC-HTP-S1)</td>
			<td>SHANGHAI ALCOTT BIOTECH CO</td>
			<td>ALC-HTP-S1</td>
			<td>Heating</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD life science</td>
			<td>R510-22</td>
			<td>Inhalant anaesthesia</td>
		</tr>
		<tr>
			<td>Matrx VIP 3000 Isofurane Vaporizer</td>
			<td>Midmark Corporation</td>
			<td>VIP 3000</td>
			<td>Anesthetization</td>
		</tr>
		<tr>
			<td>Medical nylon suture (5-0)</td>
			<td>Ningbo Medical Needle Co.</td>
			<td>5-0</td>
			<td>Close the skin</td>
		</tr>
		<tr>
			<td>Pentobarbital sodium salt</td>
			<td>Merck</td>
			<td>25MG</td>
			<td>Anesthetization</td>
		</tr>
		<tr>
			<td>Precision electronic balance</td>
			<td>Denver Instrument</td>
			<td>TB-114</td>
			<td>Weighing sensor</td>
		</tr>
		<tr>
			<td>Self-made spacer</td>
			<td></td>
			<td></td>
			<td>25-gauge needle</td>
		</tr>
		<tr>
			<td>Silk suture (5-0)</td>
			<td>Yangzhou Yuankang Medical Devices Co., Ltd.</td>
			<td>5-0</td>
			<td>Ligation</td>
		</tr>
		<tr>
			<td>Small animal microsurgery equipment</td>
			<td>Napox</td>
			<td>MA-65</td>
			<td>Surgical instruments</td>
		</tr>
		<tr>
			<td>Transmission Gel</td>
			<td>Guang Gong pai</td>
			<td>250ML</td>
			<td>Echocardiography</td>
		</tr>
		<tr>
			<td>Veet hair removal cream</td>
			<td>Reckitt Benchiser</td>
			<td>RQ/B 33 Type 2</td>
			<td>Remove hair of mice</td>
		</tr>
		<tr>
			<td>Vertical automatic electrothermal pressure steam sterilizer</td>
			<td>Hefei Huatai Medical Equipment Co.</td>
			<td>LX-B50L</td>
			<td>Auto clean the surgical instruments</td>
		</tr>
	</tbody>
</table>

operating transverse aortic constriction with absorbable suture to obtain transient myocardial hypertrophy

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate cardiomyocyte hypertrophy and slow down progression to heart failure. For novices, however, the MHP model is usually quite difficult to establish because of the technical obstacles in ventilator operation, opening the chest repeatedly, and bleeding caused by debanding. To facilitate this model, to increase the surgical success rate and to reduce the incidence of bleeding, we switched to absorbable sutures for the first TAC combing with a ventilator-free technique. Using a 2-week absorbable suture, we demonstrated that this procedure could cause significant myocardial hypertrophy in 2 weeks; and 4 weeks after surgery, myocardial hypertrophy was almost completely regressed to the baseline. Using this protocol, the operators could master the MHP model easily with a lower operation mortality.

In this study, we randomly divided 45 mice into three groups, the sham, the silk suture group, and the absorbable suture group (the number of each group on D0 (baseline), D14, and D28 after TAC was 15, 10, and 5, respectively). On D7, D14, D21, and D28 after the surgery, the constricted peak velocity was determined by echocardiography. We found that the blood flow velocity at the constriction was still greater than 3,000 mm/s in the second week after TAC even though an absorbable suture had been used to constrict the aor...

Watch this Scientific Journal Video about Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy at JoVE.com

Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy

This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning.

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate ...

Research

JoVE Journal

Medicine

A Research Method For Detecting Transient Myocardial Ischemia In Patients With Suspected Acute Coronary Syndrome Using Continuous ST-segment Analysis

Each year, an estimated 785,000 Americans will have a new coronary attack, or acute coronary syndrome (ACS). The pathophysiology of ACS involves rupture ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Murine Isolated Heart Model of Myocardial Stunning Associated with Cardioplegic Arrest

The following protocol is of use to evaluate impaired cardiac function or myocardial stunning following moderate ischemic insults. The technique is useful ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

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 ...

Full-root Aortic Valve Replacement by Stentless Aortic Xenografts in Patients with Small Aortic Roots

In patients with small aortic roots who need an aortic valve replacement with biological valve substitutes, the implantation of the stented pericardial ...

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 ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Laparoscopic Repair of Para-Esophageal Hernia Using Absorbable Biosynthetic Mesh

Para-esophageal hernia repairs are challenging procedures and there is no consensus on the optimal approach to repair. Mesh reinforcement has been ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...