Name: induction and phenotyping of acute right heart failure in a large animal model of chronic thromboembolic pulmonary hypertension
Uploaded: 2022-03-17T13:00:00.000+00:00
Duration: 7 min 41 s
Description: Watch this Scientific Journal Video about Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension at JoVE.com

This work is supported by a public grant overseen by the French National Research Agency (ANR) as part of the&#160;Investissements d&#39;Avenir Program (reference: ANR-15RHUS0002).

The study complied with the principles of laboratory animal care according to the National Society for Medical Research and was approved by the local ethic committee for animal experiments at Hospital Marie Lannelongue.
1. Chronic thromboembolic PH
<ol>
	<li>Induce chronic thromboembolic PH as previously described6,12.</li>
	<li>Briefly, induce a model of chronic thrombo-embolic PH in around 20 kg large white pigs (sus scrofa). Perform a ligature of the left pulmonary artery ligation through a left thoracotomy at week 0 (closed pericardium); and perform weekly an embolization of the right lower lobe pulmonary artery (0.2 mL to 0.4 mL per week) with a mixed solution composed with 1 mL of soft tissue glue including N-butyl-2-cyanoacrylate and 2 mL of lipidic contrast dye (lipiodol) for 5 weeks.</li>
	<li>Perform a xypho&#239;dectomy at week 0 at time of left pulmonary artery ligation to improve the echocardiography feasibility. To do this, perform a 4 cm longitudinal incision in front of the xiphoid process. Remove the xyphoid process using a diathermy knife. Close the subcutaneous plan and the skin with a running suture.</li>
	<li>Perform an additional right lower lobe pulmonary embolism at week 10 by using the same protocol explained above (step 1.2).</li>
	<li>Perform the ARHF induction (section 6) model 6 weeks after the last right lower lobe embolization (week 16) in order to avoid acute right heart lesions induced by acute pulmonary embolisms. 
	NOTE: Other large animal model of right heart failure can be used, or other pathological conditions can be induced in the chronic-thromboembolic PH model.</li>
</ol>
2. Animal positioning and catheter placements
<ol>
	<li>Perform general anesthesia as previously described6.
	<ol>
		<li>Briefly, let the animal fast for 12 h. Then perform an intramuscular injection of ketamine hydrochloride (30 mg/kg) for premedication. Perform an intravenous bolus of fentanyl (0.005 mg/kg), Propofol (2 mg/kg) and cisatracurium (0.3 mg/kg) intravenously through an ear vein and intubate non-selectively the pig with a 7 French probe.</li>
		<li>Maintain general anesthesia with inhaled 2% isoflurane, continuous infusion of fentanyl (0.004 mg/kg) and propofol (3 mg/kg).</li>
	</ol>
	</li>
	<li>After general anesthesia induction, position the pig on his back with its forelegs in a slightly spread position to allow parasternal echocardiographic acquisition (section 3).</li>
	<li>Place the device electrodes on the arms and legs (echocardiograph, workstation for hemodynamic acquisitions) prior to the placement of the sterile fields.</li>
	<li>Place an 8-French sheath into the jugular vein using the Seldinger method13.
	<ol>
		<li>Introduce an 18 G (1.3 mm x 48 mm) IV catheter into the jugular vein.
		<ol>
			<li>Perform a percutaneous puncture on the middle line at 2 cm above the manubrium with a 45&#176; orientation.</li>
			<li>After obtaining a venous reflux, insert a guidewire into the catheter (0.035 inch / 0.089 mm, 180 cm, angled).</li>
			<li>Verify the correct placement of the guidewire into the superior vena cava with fluoroscopy and dispose the 8-French sheath on the guidewire into the superior vena cava. 
			NOTE: The guidewire is correctly placed when it goes through the inferior vena cava along the right border of the spine.</li>
		</ol>
		</li>
	</ol>
	</li>
	<li>Perform a division of the right femoral vessels to introduce a fluid filled catheter into the right femoral artery for continuous systemic pressure monitoring and a balloon dilation catheter into the inferior vena cava through the femoral vein as follow.
	<ol>
		<li>Perform a 4 cm transverse incision at the groin.</li>
		<li>Place a Beckman retractor and divide the anterior face of the femoral vein and of the femoral artery using a Debackey forceps and Metzenbaum scissors.</li>
		<li>Place a 20 G catheter into the femoral artery under direct visual control and connect it to a disposable transducer with a fluid filled catheter to obtain continuous systemic blood pressure monitoring. 
		NOTE: The mean blood pressure should be continuously above 60 mmHg.</li>
		<li>Use an 18 G catheter to insert a guidewire (0.035 inch / 0.089 mm, 180 cm, angled) into the femoral vein through the inferior vena cava under fluoroscopic control.</li>
		<li>Insert a balloon dilation catheter on the guidewire through the inferior vena cava at the intrapericardial level under fluoroscopic control.</li>
	</ol>
	</li>
	<li>Perform the fluoroscopic control with a C-arm using an anteroposterior view. Place the visible markers of the balloon immediately above the diaphragm level under fluoroscopic control. Remove the guidewire when the balloon is placed.</li>
	<li>Sew a purse with a 5.0 polypropylene monofilament suture around the venous dilation balloon catheter to avoid bleeding from the femoral vein.</li>
</ol>
3. Echocardiography
<ol>
	<li>Perform the echocardiography right after the animal positioning and the catheter placement (section 2) in animals still under general anesthesia and mechanical ventilation.</li>
	<li>Acquire each echocardiographic view in cine loop format for at least 3 cardiac cycles during end-expiratory apnea.</li>
	<li>Acquire all views in 2-dimension and Tissue Doppler modes.</li>
	<li>Acquire the apical 5-chamber view under the xiphoid process.</li>
	<li>Acquire the parasternal short and long axis views on the right side of the sternum.</li>
	<li>Acquire valvular flow using continuous and pulsed Doppler modes.</li>
	<li>Acquire Tissue Doppler signals of the lateral tricuspid annulus and lateral and septal mitral annulus. 
	NOTE: Use the latest guidelines for echocardiographic assessment in humans for echocardiographic acquisitions and interpretations14.</li>
</ol>
4. Right heart catheterization
<ol>
	<li>Perform the right heart catheterization after the cardiac echo (section 3) and prior the pressure-volume loop acquisitions (section 5)</li>
	<li>Link the Swan-Ganz catheter to the disposable transducer.</li>
	<li>Introduce the Swan-Ganz catheter into the jugular 8-French sheath previously inserted into the jugular vein (section 2.4) and acquire mean right atrial, right ventricular and pulmonary artery pressures. Place the catheter under fluoroscopy if needed. 
	NOTE: Check that fluid filled catheters are well purged with saline and remove air bubbles to avoid pressure signal damping.</li>
	<li>After placing the Swan-Ganz catheter into the pulmonary artery, measure the cardiac output with the thermodilution method as explained by the manufacturer&#39;s instructions; measure simultaneously the heart rate for stroke volume calculation.
	<ol>
		<li>Ensure that the saline is at 4 &#176;C to avoid overestimation of the cardiac output.</li>
		<li>Connect the disposable transducer to the PV-loop work station for live acquisitions of pressures derived from fluid filled catheters.</li>
	</ol>
	</li>
</ol>
5. Pressure volume loop acquisition using the conductance method
NOTE: This section has been previously published15.
<ol>
	<li>Introduce the conductance catheter into the right ventricle under fluoroscopic control.
	<ol>
		<li>Verify the quality signal using &#34; in live&#34; acquisition of pressure-volume loops.</li>
	</ol>
	</li>
	<li>Activate adequate electrodes to obtain optimal signal (i.e., counter-clockwise PV-loops with physiological shape).</li>
	<li>Follow-up the pressure and volume calibration steps of the workflow according to the manufacturer&#39;s instruction (blood conductivity, parallel volume, stroke volume calibration = alpha calibration). 
	NOTE: Stroke external with the Swan-Ganz catheter can be repeated for each conditions; whereas the other calibration steps can be performed only once.</li>
	<li>Acquire PV-loop families in steady states and during acute preload reduction (i.e., acute occlusion of the inferior vena cava) during end-expiratory apnea.</li>
	<li>Perform at least 3 acquisitions per condition (steady + IVC occlusion).</li>
</ol>
6. Induction of acute right heart failure by volume and pressure overload (Figure 1).
<ol>
	<li>Induce volume overload using a 3-step saline infusion (around 2 h).
	<ol>
		<li>Start the first infusion of 15 mL/kg of saline with a free-flow infusion output.</li>
		<li>Perform the measurements (right heart catheterism, PV-loops and echocardiographic) 5 min after hemodynamic stabilization after the end of each infusion.</li>
		<li>Start the second volume infusion of 15 mL/kg immediately after the end of the measurements.</li>
		<li>Start the third volume infusion of 30 mL/kg of saline immediately after the end of the measurements. 
		CAUTION: Volume loading can induce hemodynamic compromise or pulmonary edema depending on the animal model used. In this model, volume loading revealed an adaptive response characterized by increasing cardiac output, stable right atrial pressure and preserved ventriculo-arterial coupling. 
		NOTE: Volume loading can be stopped in case of poor respiratory or hemodynamic tolerance.</li>
	</ol>
	</li>
	<li>Induce pressure overload with iterative pulmonary embolism.
	<ol>
		<li>Insert a 5 French angiographic catheter through the jugular sheath into the right lower lobe pulmonary artery under fluoroscopic control.</li>
		<li>Embolize the right lower lobe pulmonary artery with a bolus of 0.15 mL of a mixed solution composed with 1 mL of soft tissue glue including N-butyl-2-cyanoacrylate and 2 mL of lipidic contrast dye. Wash out the catheter with 10 mL of saline.</li>
		<li>Evaluate the hemodynamic response 2 minutes after the embolization using the systemic pressure and pulmonary artery pressure.</li>
		<li>Repeat embolisms of 0.15 mL every 2 minutes until obtaining hemodynamic compromise, (i.e., systolic systemic pressure &#60;90 mmHg or systolic pulmonary pressure over systolic systemic pressure ratio &#62;0.9). 
		CAUTION: Pulmonary embolism can induce severe hemodynamic compromise, sometimes irreversible, leading to immediate death. Prior starting the embolization step, be ready to start hemodynamic support (dobutamine protocol or epinephrine in case of circulatory arrest). Be ready to start PV-loops and echocardiographic monitoring. As this step may be associated with severe hemodynamic compromise, right heart catheterization using the Swan-Ganz catheter can be avoided so as to start dobutamine support sooner.</li>
	</ol>
	</li>
</ol>
7. Induce restoration of the systemic hemodynamic with dobutamine
<ol>
	<li>After reaching hemodynamic compromise and performing PV-loops and echocardiographic acquisitions, start dobutamine infusion at 2.5 &#181;g/kg/min. 
	NOTE: Other drugs or treatments can be started at this time point.</li>
	<li>Wait 10 to 15 min for hemodynamic stabilization.</li>
	<li>Perform right heart catheterization, PV-loops and echocardiographic acquisitions.</li>
	<li>Increase the dose of dobutamine infusion to 5 &#181;g/kg/min.</li>
	<li>Wait 15 min for hemodynamic stabilization and repeat acquisitions.</li>
	<li>Repeat right heart catheterization, PV-loops and echocardiographic acquisitions.</li>
	<li>Increase the dose of dobutamine infusion to 7.5 &#181;g/kg/min. 
	NOTE: Other doses, drugs or treatments can be initiated.</li>
</ol>
8. Euthanasia and cardiac tissue harvesting
<ol>
	<li>At the end of the protocol, perform a median sternotomy using an oscillating saw.</li>
	<li>Open the pericardium and inject a lethal solution of potassium chloride (0.2 g/kg).</li>
	<li>Harvest the heart; select samples of the right and left ventricular free-walls for pathological and molecular evaluations. 
	NOTE: The methods for the pathological evaluations of the right ventricle and for the statistics were previously reported5.</li>
</ol>

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The authors have nothing to disclose.

We describe a method to model major pathophysiological features of ARHF on chronic PH in a large animal model including volume and pressure overload and hemodynamic restoration with dobutamine. We also reported how to acquire hemodynamic and imaging data to phenotype the dynamic changes of the right ventricle at each condition created during the protocol. These methods can provide background data to build up future research protocols in the field of ARHF, particularly regarding fluid management and inotropic support.
Inducing hemodynamic compromise was a critical step in the model because of the risk of unexpected and immediate death of the animal. Consequently, we recommend inducing progressive pulmonary embolism with small embolus volumes. At time of pulmonary embolism, the investigators should be ready to immediately start data acquisitions and hemodynamic support. In our experience, we were able to realize the PV-loop acquisitions and the echocardiography prior to starting dobutamine in 6 out of 7 animals in whom the protocol was completed.
The critical step to phenotype the right ventricle is to obtain comprehensive hemodynamic, PV loop and echocardiographic data. Right heart catheterization allows one to estimate cardiac output and stroke volume changes for each condition. Changes in cardiac output and stroke volume can be further evaluated with echocardiography. This multimodal analysis of cardiac output and stroke volume changes better the external volume calibration of the PV-loops. Importantly, absolute values and rates of changes of PV-loop parameters can be more precisely quantified by including cardiac output and stroke volume changes with external methods performed for each situation.
We observed that volume loading did not induce hemodynamic compromise but rather revealed the adaptive phenotype of the PH model as we observed an increase in cardiac output, stroke volume and systemic pressure with preserved ventriculo-arterial coupling. Therefore, in our model, initial volume loading provided the conditions to observe a major drop in cardiac output and stroke volume after acute pulmonary embolism, hence increasing the sensitivity of the model. Future studies should determine the effect of volume loading or fluid depletion at the time of hemodynamic compromise.
Our protocol has several limitations. This protocol was not built to analyze the cause of the edema, but it may represent an interesting research area. Another limit of the protocol is the time-consumption and the skills required to perform all the steps. The volume loading phase can be shortened or removed from the protocol, but this may result in a lower decrease in the absolute value of the cardiac output and stroke volume after acute pulmonary embolisms. The skills required to perform the protocol require the collaboration of several investigators to place the catheter under fluoroscopy, perform the echocardiography, and analyze in real time the PV-loop quality. We acknowledge that we did not performed 3-dimensional evaluations of the RV volumes. We aim to develop 3-dimensional evaluations of RV volumes as it may provide more precision in the RV volume calibration for RV PV-loop evaluations. One of the first steps would be to evaluate the feasibility of the method. Furthermore, our protocol requires specific facilities such as an operating room and fluoroscopy for invasive RV evaluations.
To our knowledge, we have described the first animal model of ARHF with chronic PH. Previous studies reported dynamic changes of the right ventricle with dobutamine and levosimendan after acute pulmonary artery constriction7. In our group, we also quantified the RV reserve using dobutamine infusion in chronic PH without hemodynamic compromise15. Multibeat PV-loops are considered the gold standard method to quantify the end-systolic elastance, which represents the ventricular contractility independently from loading conditions16. RV elastance (Ees=end systolic elastance) absolute values should be interpreted with caution as there are several methodological limits. The main limits are the definition of the end-systolic point and the precision of the volume calibration with external methods (thermodilution and echocardiography)17. The ratio of end-systolic elastance over arterial elastance (Ea=end-systolic pressure over stroke volume ratio), known as the ventriculo-arterial coupling (Ees/Ea) ratio, reduces the errors due to external volume calibration. The ventriculo-arterial coupling is of major interest in the field of pulmonary hypertension as it captures the adaptation of RV contractility to increased afterload. Methods measuring RV adaptation to afterload have gained major interest in recent years because it has better phenotyping of patients with PH18,19,20.
Our methods provided values of ventriculo-arterial coupling (i.e., Ees/Ea) consistent with previously published values21 and with RV function estimation using echocardiography. In this protocol, we show that acute vena cava occlusion is safe when performed in the context of hemodynamic compromise. Moreover, RV echocardiographic evaluation in the large animal model was complementary from RV echocardiographic evaluation in small animals models as it allowed to quantify different RV function parameters compared with previously reported mice models with RV remodeling22.
The methods described in this study can be used for different research protocols aiming to address key questions in the field of ARHF. First, these methods can be used to perform research protocols aiming to compare different treatment strategies in the context of ARHF on chronic PH. Secondly, iterative and simultaneous PV-loop and echocardiographic evaluation can allow to validate echocardiographic indices in different situations of clinical interest.

Acute right heart failure (ARHF) has been recently defined as a rapidly progressive syndrome with systemic congestion resulting from impaired right ventricular (RV) filling and/or reduced RV flow output1. ARHF may occur in several conditions such as left-sided heart failure, acute pulmonary embolism, acute myocardial infarction or pulmonary hypertension (PH). In the case of PH, ARHF onset is associated with a 40% risk of short-term mortality or urgent lung transplantation2,3,4. Here, we describe how to create a large animal model of ARHF in the setting of chronic pulmonary hypertension and how to evaluate the right ventricle using echocardiography and pressure-volume loops.
Pathophysiological features of ARHF include RV pressure overload, volume overload, a decrease in RV output, an increase in central venous pressure and/or a decrease in systemic pressure. In chronic PH, there is an initial increase in RV contractility allowing to preserve cardiac output despite the increase in pulmonary vascular resistance. Therefore, in the context of ARHF on chronic PH, the right ventricle can generate nearly isosystemic pressures, particularly under inotropic support. Taken together, ARHF on chronic PH and hemodynamic restoration with inotropes lead to the development of acute RV ischemic lesions, as recently described in our large animal model5. The increase in inotropes creates an increased energetic demand that may further develop ischemic lesions, and finally lead to the development of end-organ dysfunction and poor clinical outcomes. However, there is no consensus about how to manage patients with ARHF on PH, mainly regarding fluid management, inotropes and the role of extra-corporeal circulatory support. Consequently, a large animal model of acute right heart failure may help to provide pre-clinical data on ARHF clinical management.
As a first step to quantify the response to therapy, simple and reproducible methods to phenotype the right ventricle are needed. To date, there is no consensus about how to better phenotype the RV morphology and function of patients with ARHF. The gold standard method to evaluate RV contractility (i.e., intrinsic capacity to contract) and ventriculo-arterial coupling (i.e., contractility normalized by ventricular afterload; an index of ventricular adaptation) is the analysis of pressure-volume (PV) loops. This method is twice invasive because it requires right heart catheterization and a transient reduction in RV preload using a balloon inserted in the inferior vena cava. In clinical practice, non-invasive and repeatable methods to evaluate the right ventricle are needed. Cardiac magnetic resonance (CMR) is considered as the gold standard for non-invasive evaluation of the right ventricle. In patients with ARHF on chronic PH who are managed in intensive care unit (ICU), the use of CMR may be limited because of the patient&#39;s unstable hemodynamic condition; moreover, repeated CMR evaluations, several times a day, including at night, may be limited because of its cost and limited availability. Conversely, echocardiography allows non-invasive, reproducible and low-cost RV morphology and function evaluations in ICU patients.
Large animal models are ideal to perform preclinical studies focusing on the relationship between invasive hemodynamic parameters and non-invasive parameters. The large white pig anatomy is close to humans. Consequently, most of the echocardiographic parameters described in humans are quantifiable in pigs. Some minor variations exist between human and pig heart that must be taken into account for echocardiographic studies. Pigs present a constitutional dextrocardia and a slightly counterclockwise rotation of the heart axis. As a result, the apical 4-chamber view becomes an apical 5-chamber view and the acoustic window is situated below the xiphoid appendix. Additionally, parasternal long and short axis views acoustic windows are situated on the right side of the sternum.
Here, we describe a novel method to induce ARHF in a large animal model of chronic thromboembolic PH and to restore hemodynamic using dobutamine. We also report RV ischemic lesions present in the model within 2&#8722;3 hours after hemodynamic restoration with dobutamine. Moreover, we describe how to acquire RV PV-loops and echocardiographic RV parameters at each condition providing insights on the dynamic changes in RV morphology and function. As the large animal model of chronic thromboembolic PH and the PV-loop methods were previously described6, these sections will be briefly described. Also, we reported results of echocardiographic evaluations which are deemed potentially difficult in porcine models. We will explain the methods to achieve repeated echocardiographic in the model.
The model of ARHF on chronic PH reported in this study can be used to compare different therapeutic strategies. The methods of RV phenotyping can be used in other large animal models mimicking clinically relevant situations such as acute pulmonary embolism7, RV myocardial infarction8, acute respiratory distress syndrome9 or right heart failure associated with left ventricular failure10 or left ventricular mechanical circulatory support11.

<table><tbody><tr><td>Radiofocus Introducer II</td><td>Terumo</td><td>RS+B80K10MQ</td><td>catheter sheath</td></tr><tr><td>Equalizer, Occlusion Ballon Catheter</td><td>Boston Scientific</td><td>M001171080</td><td>ballon for inferior vena cava occlusion</td></tr><tr><td>Guidewire</td><td>Terumo</td><td>GR3506</td><td>0.035; angled</td></tr><tr><td>Vigilance monitor</td><td>Edwards</td><td>VGS2V</td><td>Swan-Ganz associated monitor</td></tr><tr><td>Swan-Ganz</td><td>Edwards</td><td>131F7</td><td>Swan-Ganz catheter 7 F; usable lenghth 110 cm</td></tr><tr><td>Echocardiograph; Model: Vivid 9</td><td>General Electrics</td><td>GAD000810 and H45561FG</td><td>Echocardiograph</td></tr><tr><td>Probe for echo, M5S-D</td><td>General Electrics</td><td>M5S-D</td><td>Cardiac ultrasound transducer</td></tr><tr><td>MPVS-ultra Foundation system</td><td>Millar</td><td>PL3516B49</td><td>Pressure-volume loop unit; includes a powerLab16/35, MPVS-Ultra PV Unit, bioamp and bridge amp and cables</td></tr><tr><td>Ventricath 507</td><td>Millar</td><td>VENTRI-CATH-507</td><td>conductance catheter</td></tr><tr><td>Lipiodol ultra-fluid</td><td>Guerbet</td><td>306 216-0</td><td>lipidic contrast dye</td></tr><tr><td>BD Insyte Autoguard</td><td>Becton, Dickinson and Company</td><td>381847</td><td>IV catheter</td></tr><tr><td>Arcadic Varic</td><td>Siemens</td><td>A91SC-21000-1T-1-7700</td><td>C-arm</td></tr><tr><td>Prolene 5.0</td><td>Ethicon</td><td>F1830</td><td>polypropilene monofil</td></tr></tbody></table>

induction and phenotyping of acute right heart failure in a large animal model of chronic thromboembolic pulmonary hypertension

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The morphological and functional phenotyping of the right ventricle is of particular importance in the context of hemodynamic compromise in patients with ARHF. Here, we describe a method to induce ARHF in a previously described large animal model of chronic PH, and to phenotype, dynamically, right ventricular function using the gold standard method (i.e., pressure-volume PV loops) and with a non-invasive clinically available method (i.e., echocardiography). Chronic PH is first induced in pigs by left pulmonary artery ligation and right lower lobe embolism with biological glue once a week for 5 weeks. After 16 weeks, ARHF is induced by successive volume loading using saline followed by iterative pulmonary embolism until the ratio of the systolic pulmonary pressure over systemic pressure reaches 0.9 or until the systolic systemic pressure decreases below 90 mmHg. Hemodynamics are restored with dobutamine infusion (from 2.5 &#181;g/kg/min to 7.5 &#181;g/kg/min). PV-loops and echocardiography are performed during each condition. Each condition requires around 40 minutes for induction, hemodynamic stabilization and data acquisition. Out of 9 animals, 2 died immediately after pulmonary embolism and 7 completed the protocol, which illustrates the learning curve of the model. The model induced a 3-fold increase in mean pulmonary artery pressure. The PV-loop analysis showed that ventriculo-arterial coupling was preserved after volume loading, decreased after acute pulmonary embolism and was restored with dobutamine. Echocardiographic acquisitions allowed to quantify right ventricular parameters of morphology and function with good quality. We identified right ventricular ischemic lesions in the model. The model can be used to compare different treatments or to validate non-invasive parameters of right ventricular morphology and function in the context of ARHF.

Feasibility 
We describe the results of 9 consecutive procedures of ARHF induction in a large animal CTEPH model previously reported5. The duration of the protocol was around 6 hours to complete, including anesthesia induction, installation, vascular access/catheter placements, induction of volume/pressure overload and hemodynamic restoration, data acquisitions and euthanasia. Each hemodynamic condition requires around 40 minutes to achieve induction of the condition, hemodynamic stabilization and data acquisitions.
The protocol was achieved in 7 out of 9 animals, which represents the learning curve. Three additional protocols were successfully achieved after these described (not published). The cause of the 2 protocol failures was the induction of an irreversible hemodynamic failure after the pulmonary embolism phase.
PV loops were not acquired in 1 out of 7 animals at time of hemodynamic compromise because of the necessity to provide rapid systemic hemodynamic restoration with an epinephrine bolus after the right heart catheterism and the cardiac echo. In this case, dobutamine was started immediately after the restoration of the systemic hemodynamic with epinephrine.
Effects of volume and pressure overload on hemodynamics and RV function 
Acute volume loading did not induce ARHF but rather highlighted the adaptive phenotype of the chronic PH model. With volume loading, the cardiac output increased without increase in right atrial pressure, while the ventriculo-arterial coupling remained stable (Figure 2).
Hemodynamic compromise criteria were achieved after 1 embolus in 1 animal, 2 emboli in 2 animals, 3 emboli in 5 animals and 4 emboli in 1 animal. Two animals died immediately after PE (1 animal with 1 embolus and 1 animal with 4 emboli). In another animal, severe hypotension required an epinephrine bolus and immediate starting of dobutamine prior to PV-loop and echocardiographic data acquisitions. The 2 deaths occurring immediately after acute pulmonary embolism were associated with acute thrombosis of the right heart cavities (as illustrated in Figure 3).
Hemodynamic compromise was associated with a significant decrease in cardiac output, stroke volume and ventriculo-arterial coupling (Ees/ea), whereas RV contractility remained stable (Figure 2); there was a two-fold increase in right atrial pressure and mean pulmonary artery pressure.
Dobutamine effect on ARHF 
Dobutamine restored a cardiac output, stroke volume, and ventriculo-arterial coupling within normal range (Figure 2).
Echocardiography 
Echocardiography was feasible providing quantification of dynamic changes in RV size and function during the protocol (Figure 4). Echocardiographic parameters were not assessed in 1 animal with severe hemodynamic compromise after pulmonary embolism requiring an epinephrine bolus and immediate starting of dobutamine.
RV PV-loops 
Pressure volume loop analysis allowed dynamic quantification of RV end-systolic elastance and ventriculo-arterial coupling (Figure 2 and Figure 5).
Right ventricular ischemic lesions 
After hematein, eosin, and saffron staining, we observed RV ischemic lesions in the subendocardial and in the subepicardial layers of the RV free-wall (Figure 6). The ischemic lesions were characterized by clusters of hypereosinophilic cardiomyocytes with picnotic nucleus.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58057/58057fig1.jpg" /> 
Figure 1: Protocol summary. PH, pulmonary hypertension; VL1, volume loading with 15 mL/kg of saline; VL2, 15 mL/kg of saline; VL3, 30 mL/kg of saline; ARHF, acute right heart failure; PE, pulmonary embolism. *systemic systolic pressure &#60;90 mmHg or systolic pulmonary/systemic pressures ratio &#62;0.9. This figure has been modified from5. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58057/58057fig2.jpg" /> 
Figure 2: Individual hemodynamic and pressure-volume loop dynamic changes. MPAP, mean pulmonary artery pressure; MAP, mean arterial pressure; RAP, right atrial pressure; HR, heart rate; SV, stroke volume; CO, cardiac output; Ees; right ventricular end-systolic elastance; Ea, arterial elastance. Plots are median and interquartile range. *P&#60;0.05 compared to baseline; comparisons were performed using Wilcoxon matched-pairs signed rank tests with GraphPad Prism 6. This figure has been modified from5. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58057/58057fig3.jpg" /> 
Figure 3: Example of cause of protocol failure: acute right heart thrombosis (arrow) after pulmonary embolism responsible for irreversible hemodynamic compromise, immediate death and protocol failure. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58057/58057fig4.jpg" /> 
Figure 4: Representative echocardiographic windows and results.&#160;(A) Position for acquisition of the apical 5-chamber (A5C) view. (B) Position for acquisition of the parasternal short axis (PSSAX) view. (C) Dynamic echocardiographic evaluations of the A5C and the PSSAX views during the different steps of the protocol. VL, volume loading; PE, pulmonary embolism; Dobu 2.5, dobutamine 2.5 &#181;g/kg/min; Dobu 7.5, dobutamine 7.5 &#181;g/kg/min. *right ventricle; **left ventricle. This figure has been modified from5. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58057/58057fig5.jpg" /> 
Figure 5: Representative dynamic RV multibeat pressure-volume loops. PH, pulmonary hypertension; PE, pulmonary embolism; Ees, end-systolic elastance (black line labelled *); Ea, arterial elastance (black line labelled **); Ees/Ea, ventriculo-arterial coupling. This figure has been modified from5. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig5large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/58057/58057fig6.jpg" /> 
Figure 6: Representative RV ischemic lesions in the subendocardium and in the sub epicardium layers. (A) Subepicardial ischemic lesion; (B) Subendocardial ischemic lesions; (C) Magnification of a border of a subepicardial ischemic lesion with normal nuclei (1), intracytoplasmic vacuolization (2) and pyknotic nuclei (3). (D) individual numbers of subendocardic and subepicardic ischemic lesions in 2 cm length samples of RV free-wall from animals with acute right heart failure (ARHF) on chronic pulmonary hypertension (PH), animals with chronic PH and healthy controls; plots are medians. Comparisons were performed using Mann-Whitney test with GraphPad Prism 6. *P&#60;0.05. This figure has been modified from5. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/58057/58057fig6large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

Watch this Scientific Journal Video about Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension at JoVE.com

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure pathophysiology.

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

induction-phenotyping-acute-right-heart-failure-large-animal-model

Nanyang Technological University

Research

JoVE Journal

Medicine

2.9K Views.  Marie Lannelongue Hospital. Dit model van acuut rechterhartfalen in de context van coronaire pulmonale hypertensie kan helpen om de pathofysiologie van deze klinisch relevante situatie beter te begrijpen. Inductie van acuut rechterhartfalen door middel van volume- en drukoverbelasting is gemakkelijk reproduceerbaar en dupliceert de belangrijkste pathofysiologische aspecten van de overeenkomstige klinische situatie. Na bevestiging van een geschikt niveau van sedatie, plaatst u een centraal veneuze katheter door de halsad...

Video: Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle proliferation and obliteration of pulmonary arterioles. This in turn results in right ventricular (RV) failure, with significant morbidity and mortality. Rodent models of PAH, in the mouse and the rat, are important for understanding the pathophysiology underlying this rare disease. Notably, different models of PAH may be associated with different degrees of pulmonary hypertension, RV hypertrophy and RV failure. Therefore, a complete hemodynamic characterization of mice and rats with PAH is critical in determining the effects of drugs or genetic modifications on the disease. 
Here we demonstrate standard procedures for assessment of right ventricular function and hemodynamics in both rat and mouse PAH models. Echocardiography is useful in determining RV function in rats, although obtaining standard views of the right ventricle is challenging in the awake mouse. Access for right heart catheterization is obtained by the internal jugular vein in closed-chest mice and rats. Pressures can be measured using polyethylene tubing with a fluid pressure transducer or a miniature micromanometer pressure catheter. Pressure-volume loop analysis can be performed in the open chest. After obtaining hemodynamics, the rodent is euthanized. The heart can be dissected to separate the RV free wall from the left ventricle (LV) and septum, allowing an assessment of RV hypertrophy using the Fulton index (RV/(LV+S)). Then samples can be harvested from the heart, lungs and other tissues as needed.

Pulmonary arterial hypertension (PAH) is a disease of pulmonary arterioles that leads to their obliteration and the development of right ventricular failure. Rodent models of PAH are critical in understanding the pathophysiology of PAH. Here we demonstrate hemodynamic characterization, with right heart catheterization and echocardiography, in the mouse and rat.

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle ...

A Large Animal Model for Pulmonary Hypertension and Right Ventricular Failure: Left Pulmonary Artery Ligation and Progressive Main Pulmonary Artery Banding in Sheep

Decompensated right ventricular failure (RVF) in pulmonary hypertension (PH) is fatal, with limited medical treatment options. Developing and testing novel therapeutics for PH requires a clinically relevant large animal model of increased pulmonary vascular resistance and RVF. This manuscript discusses the latest development of the previously published ovine PH-RVF model that utilizes left pulmonary artery (PA) ligation and main PA occlusion. This model of PH-RVF is a versatile platform to control not only the disease severity but also the RV&#39;s phenotypic response.
Adult sheep (60-80 kg) underwent left PA (LPA) ligation, placement of main PA cuff, and insertion of RV pressure monitor. PA cuff and RV pressure monitor were connected to subcutaneous ports. Subjects underwent progressive PA banding twice per week for 9 weeks with sequential measures of RV pressure, PA cuff pressures, and mixed venous blood gas (SvO2). At the initiation and endpoint of this model, ventricular function and dimensions were assessed using echocardiography. In a representative group of 12 animal subjects, RV mean and systolic pressure increased from 28 &#177; 5 and 57 &#177; 7 mmHg at week 1, respectively, to 44 &#177; 7 and 93 &#177; 18 mmHg (mean &#177; standard deviation) by week 9. Echocardiography demonstrated characteristic findings of PH-RVF, notably RV dilation, increased wall thickness, and septal bowing. The longitudinal trend of SvO2 and PA cuff pressure demonstrates that the rate of PA banding can be titrated to elicit varying RV phenotypes. A faster PA banding strategy led to a precipitous decline in SvO2 &#60; 65%, indicating RV decompensation, whereas a slower, more paced strategy led to the maintenance of physiologic SvO2 at 70%-80%. One animal that experienced the accelerated strategy developed several liters of pleural effusion and ascites by week 9. This chronic PH-RVF model provides a valuable tool for studying molecular mechanisms, developing diagnostic biomarkers, and enabling therapeutic innovation to manage RV adaptation and maladaptation from PH.

This manuscript describes the surgical technique and experimental approach to develop severe right ventricular pressure overload to model their adaptive and maladaptive phenotypes.

Decompensated right ventricular failure (RVF) in pulmonary hypertension (PH) is fatal, with limited medical treatment options. Developing and testing ...

Bioengineering

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to pulmonary hypertension. For a better understanding and treatment of this disease, precise hemodynamic monitoring of left and right ventricular parameters is important. For this reason, it is essential to establish experimental pig models of cardiac hemodynamics and measurements for research purpose. 
This article shows the induction of ARDS by using oleic acid (OA) and consequent right ventricular dysfunction, as well as the instrumentation of the pigs and the data acquisition process that is needed to assess hemodynamic parameters. To achieve right ventricular dysfunction, we used oleic acid (OA) to cause ARDS and accompanied this with pulmonary artery hypertension (PAH). With this model of PAH and consecutive right ventricular dysfunction, many hemodynamic parameters can be measured, and right ventricular volume load can be detected. 
All vital parameters, including respiratory rate (RR), heart rate (HR) and body temperature were recorded throughout the whole experiment. Hemodynamic parameters including femoral artery pressure (FAP), aortic pressure (AP), right ventricular pressure (peak systolic, end systolic and end diastolic right ventricular pressure), central venous pressure (CVP), pulmonary artery pressure (PAP) and left arterial pressure (LAP) were measured as well as perfusion parameters including ascending aortic flow (AAF) and pulmonary artery flow (PAF). Hemodynamic measurements were performed using transcardiopulmonary thermodilution to provide cardiac output (CO). Furthermore, the PiCCO2 system (Pulse Contour Cardiac Output System 2) was used to receive parameters such as stroke volume variance (SVV), pulse pressure variance (PPV), as well as extravascular lung water (EVLW) and global end-diastolic volume (GEDV). Our monitoring procedure is suitable for detecting right ventricular dysfunction and monitoring hemodynamic findings before and after volume administration.

We present a protocol of creating right ventricular dysfunction in a pig model by inducing ARDS. We demonstrate invasive monitoring of left and right ventricular cardiac output using flow probes around the aorta and the pulmonary artery, as well as blood pressure measurements in the aorta and pulmonary artery.

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

Induction and Characterization of Pulmonary Hypertension in Mice using the Hypoxia/SU5416 Model

Pulmonary Hypertension (PH) is a pathophysiological condition, defined by a mean pulmonary arterial pressure exceeding 25 mm Hg at rest, as assessed by right heart&#160;catheterization. A broad spectrum of diseases can lead to PH, differing in their etiology, histopathology, clinical presentation, prognosis, and response to&#160;treatment. Despite significant progress in the last years, PH remains an uncured disease. Understanding the underlying mechanisms can pave the way for the development of new therapies. Animal models are important research tools to achieve this goal. Currently, there are several models available for recapitulating PH. This protocol describes a two-hit mouse PH model. The stimuli for PH development are hypoxia and the injection of SU5416, a vascular endothelial growth factor (VEGF) receptor antagonist. Three weeks after initiation of Hypoxia/SU5416, animals develop pulmonary vascular remodeling imitating the histopathological changes observed in human PH (predominantly Group 1). Vascular remodeling in the pulmonary circulation results in the remodeling of the right&#160;ventricle (RV). The procedures for measuring RV pressures (using the open chest method), the morphometrical analyses of the RV (by dissecting and weighing both cardiac ventricles) and the histological assessments of the remodeling (both pulmonary by assessing vascular remodeling and cardiac by assessing RV cardiomyocyte hypertrophy and fibrosis) are described in detail. The advantages of this protocol are the possibility of the application both in wild type and in genetically modified mice, the relatively easy and low-cost implementation, and the quick development of the disease of interest (3 weeks). Limitations of this method are that mice do not develop a severe phenotype and PH is reversible upon return to normoxia. Prevention, as well as therapy studies, can easily be implemented in this model, without the necessity of advanced skills (as opposed to surgical rodent models).

This protocol describes the induction of pulmonary hypertension (PH) in mice based on the exposure to hypoxia and the injection of a VEGF receptor antagonist. The animals develop PH and right ventricular (RV) hypertrophy 3 weeks after the initiation of the protocol. The functional and morphometrical characterization of the model is also presented.

Pulmonary Hypertension (PH) is a pathophysiological condition, defined by a mean pulmonary arterial pressure exceeding 25 mm Hg at rest, as assessed by ...

Chronic Ovine Model of Right Ventricular Failure and Functional Tricuspid Regurgitation

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to suboptimal clinical results. We set out to establish a chronic ovine model of FTR and right heart failure to investigate the mechanisms of FTR. Twenty adult male sheep (6-12 months old, 62 &#177; 7 kg) underwent a left thoracotomy and baseline echocardiography. A pulmonary artery band (PAB) was placed and cinched around the main pulmonary artery (PA) to at least double the systolic pulmonary artery pressure (SPAP), inducing right ventricular (RV) pressure overload and signs of RV dilatation. PAB acutely increased the SPAP from 21 &#177; 2 mmHg to 62 &#177; 2 mmHg. The animals were followed for 8 weeks, symptoms of heart failure were treated with diuretics, and surveillance echocardiography was used to assess for pleural and abdominal fluid collection. Three animals died during the follow-up period due to stroke, hemorrhage, and acute heart failure. After 2 months, a median sternotomy and epicardial echocardiography were performed. Of the surviving 17 animals, 3 developed mild tricuspid regurgitation, 3 developed moderate tricuspid regurgitation, and 11 developed severe tricuspid regurgitation. Eight weeks of pulmonary artery banding resulted in a stable chronic ovine model of right ventricular dysfunction and significant FTR. This large animal platform can be used to further investigate the structural and molecular basis of RV failure and functional tricuspid regurgitation.

Right ventricular failure and functional tricuspid regurgitation are associated with left-sided heart disease and pulmonary hypertension, which contribute significantly to morbidity and mortality in patients. Establishing a chronic ovine model to study right ventricular failure and functional tricuspid regurgitation will help in understanding their mechanisms, progression, and possible treatments.

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to ...

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of RVF. Previous rat models imitating RVF progression have been described. Compared with rats, mice are more accessible, economical, and widely used in animal experiments. We developed a pulmonary artery constriction (PAC) approach which is comprised of banding the pulmonary trunk in mice to induce right ventricular (RV) hypertrophy. A special surgical latch needle was designed that allows for easier separation of the aorta and the pulmonary trunk. In our experiments, the use of this fabricated latch needle reduced the risk of arteriorrhexis and improved the surgical success rate to 90%. We used different padding needle diameters to precisely create quantitative constriction, which can induce different degrees of RV hypertrophy. We quantified the degree of constriction by evaluating the blood flow velocity of the PA, which was measured by noninvasive transthoracic echocardiography. RV function was precisely evaluated by right heart catheterization at 8 weeks after surgery. The surgical instruments made inhouse were composed of common materials using a simple process that is easy to master. Therefore, the PAC approach described here is easy to imitate using instruments made in the lab and can be widely used in other labs. This study presents a modified PAC approach that has a higher success rate than other models and an 8-week postsurgery survival rate of 97.8%. This PAC approach provides a useful technique for studying the mechanism of RVF and will enable an increased understanding of RVF.

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of ...

Biology

A Porcine Model of Acute Autologous Pulmonary Embolism

Acute pulmonary embolism (PE) is a potentially life-threatening condition that causes abrupt obstruction of the pulmonary arteries, leading to acute right heart failure. Novel diagnostic methods and catheter-directed therapies are being developed rapidly, and there is an obvious need for a realistic PE animal model that can be used for pathophysiological evaluation and preclinical testing.
This protocol introduces a porcine model employing large autologous pulmonary emboli. Instrumentations are performed with minimally invasive techniques, creating a close-chest model that enables the investigation of various treatment options with high reproducibility. Three hours after drawing blood to create autologous emboli ex vivo, the induction of PE caused an immediate increase in the mean pulmonary arterial pressure (17&#160;&#177; 3 mmHg to 33&#160;&#177; 6 mmHg, p &#60; 0.0001) and heart rate (50&#160;&#177; 9 beats&#183;min-1 to 63&#160;&#177; 6 beats&#183;min-1,&#160;p &#60; 0.0003) accompanied by a decreased cardiac output (5.0&#160;&#177; 0.8 L/min to 4.5&#160;&#177; 0.9 L/min, p &#60; 0.037) compared to baseline. The CT pulmonary angiography revealed multiple emboli, and the pulmonary obstruction percentage was increased compared to baseline (0% [0-0] to 57.1% [38.8-63.3], p &#60; 0.0001). In the acute phase, the phenotype is comparable to intermediate-risk PE.
The model represents a realistic and well-characterized phenotype of intermediate-risk PE and creates an opportunity to test novel diagnostic methods, interventional and pharmaceutical treatments, and hands-on training for healthcare workers in interventional procedures.

This study presents a porcine model of pulmonary embolism (PE) using large autologous emboli that replicate acute intermediate-risk PE. The model is well-suited for the evaluation of both pathophysiology and treatment responses.

Acute pulmonary embolism (PE) is a potentially life-threatening condition that causes abrupt obstruction of the pulmonary arteries, leading to acute right ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

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