Method Article
The goal of this protocol is to demonstrate a porcine exsanguination cardiac arrest model and a specifically built selective aortic arch perfusion circuit for translational research.
Hemorrhage constitutes the majority of potentially preventable deaths from trauma. There is growing interest in endovascular resuscitation techniques such as selective aortic arch perfusion (SAAP) for patients in cardiac arrest. This involves active perfusion of the coronary circulation via a thoracic aortic balloon catheter and is approaching clinical application. However, the technique is complex and requires refinement in animal models before human use can be considered. This paper describes a large animal model of exsanguination cardiac arrest treated with a bespoke SAAP system.
Swine were anesthetized, instrumented and a splenectomy was performed before a controlled, logarithmic exsanguination was initiated. Animals were heparinized and the shed blood collected in a reservoir. Once cardiac arrest was observed, the blood was pumped through an extra-corporeal circuit into an oxygenator and then delivered through a 10 Fr balloon catheter placed in the thoracic aorta.
This resulted in the return of a spontaneous circulation (ROSC) as demonstrated by ECG and aortic root pressure waveform. This model and accompanying SAAP system allow for standardized and reproducible recovery from exsanguination cardiac arrest.
Hemorrhage accounts for the majority of potentially preventable trauma deaths1. In the terminal stages of exsanguination, coronary perfusion is reduced, leading to cardiac arrest and death. Current strategies – intravenous transfusion and cardiac massage – are ineffective as they do not address the failure of coronary perfusion.
SAAP is a catheter-based resuscitation technique that aims to address this problem by the infusion of oxygenated resuscitation fluid and drugs directly to proximal aorta, perfusing the coronary and cerebral circulation. Limited swine studies have demonstrated promising outcomes in restoring cardiac activity following ventricular fibrillation and hemorrhagic cardiac arrest2,3,4. However, SAAP research is ongoing and the technique remains in the pre-clinical phases.
There are several technical challenges with SAAP. It is critical that a certain volume of perfusate be delivered via the catheter at a precise infusion rate, and currently there is no commercially available, FDA approved catheter for use in SAAP. The technique requires a specific circuit which is capable of efficiently storing, oxygenating and delivering perfusate during SAAP. The aim of this study is to present a traumatic pulseless electrical activity (PEA) cardiac arrest animal model and custom built, reliable SAAP system for use in exploring this tool in exsanguination animal research.
This study was conducted at the Medical School Teaching Facility (MSTF, University of Maryland, Baltimore, MD, USA), which is accredited by the American Association for Laboratory Animal Science. The study protocol was approved by the local Institutional Animal Care and Use Committee.
1. Animal selection and housing
2. Sedation and induction of general anesthesia
3. Surgery
4. Instrumentation
NOTE: See Table 1 for key steps in connecting the SAAP circuit.
5. Exsanguination
6. SAAP
7. Peripheral perfusion
NOTE: Following successful SAAP resuscitation depending on the study protocol, further volume replacement can be continued peripherally using the SAAP circuit.
8. Euthanasia
Aortic root blood pressure was 83/58 mmHg at baseline and gradually decreased to 0-10 mmHg during the exsanguination. Following onset of pulseless electrical activity (PEA), SAAP was performed, during which, the systolic blood pressure rapidly increased to 120 mmHg for the duration of SAAP (Figure 4). Following cessation of SAAP and aortic balloon deflation BSP dropped to about 60 mmHg however it gradually increased again during the post- SAAP period to baseline levels with a couple of spikes representing IV fluid bolus (arrows).
Figure 1: Diagram demonstrating access sites in the swine for data collection and monitoring. 7 Fr sheath placed in the left Internal Jugular Vein (IJV) can used for drug delivery depending on the protocol needs. A 7 Fr Sheath in the right IJV allows placement of a pressure transducer into the right atrium, and a 7 Fr Sheath in the right Carotid artery allows placement of a pressure transducer into the aortic arch. Please click here to view a larger version of this figure.
Figure 2: Diagram of the SAAP circuit demonstrating the setup of the circuit elements including blood reservoir, oxygenator, centrifugal and peristaltic pumps as well as the reperfusion limb, main perfusion limb, SAAP limb and peripheral limb. *Exsanguination tubing not shown Please click here to view a larger version of this figure.
Figure 3: Diagram demonstrating the SAAP circuit elements: SAAP perfusion limb and peripheral perfusion limbs with percutaneous access and SAAP catheter set up in a swine model. Aortic valve (AV) closure bolus and SAAP balloon inflation bolus syringes are demonstrated to be connected to the corresponding ports in the SAAP catheter. A 15 Fr ECMO cannula placed in the right femoral artery is connected to exsanguination tubing. Please click here to view a larger version of this figure.
Figure 4: Trend in the aortic root blood pressure (BP) throughout the exsanguination, during selective aortic arch perfusion (SAAP) following cardiac arrest and following SAAP. Dotted line demonstrating a significant temporary reduction in SBP upon SAAP balloon deflation. Please click here to view a larger version of this figure.
Key Steps | Key Components | |
Construct the circuit tubing using barbed Y and straight connectors to incorporate a reperfusion limb, main perfusion limb, SAAP perfusion limb and a peripheral perfusion limb (Figure 1) | 3/8” ID tubing, Barbed Connectors | |
Secure the connectors with cable ties | Cable ties | |
Connect the proximal tubing to the reservoir | Blood Reservoir | |
Connect the tubing to the centrifugal pump | Centrifugal pump | |
Connect the tubing to the oxygenator | Oxygenator | |
Connect the oxygenator to oxygen source | Oxygen source, Gas tubing | |
Load the tubing into the peristaltic pump head | Peristaltic pump | |
Connect the SAAP perfusion limb to the SAAP catheter and the peripheral perfusion limb to the venous catheter (Figure 2) | SAAP catheter Venous catheter 3-way and 2-way stopcocks |
Table 1: Key steps and components of constructing the SAAP circuit.
Adequate perfusate oxygenation is a critical capability of SAAP12. We use a filter that is integrated with a reservoir. The filter is connected to an oxygen cylinder via standard oxygen tubing. The oxygen flow is delivered to the oxygenator at 6 L/min. The centrifugal pump incorporated in the circuit propels the blood, which is filtered through the oxygenator. Adequate oxygenation can be confirmed by performing a blood gas analysis of a sample from the perfusion limb of the circuit. A blood gas sample performed during an experiment using the model confirmed sufficient oxygenation with findings of: pO2 746 mmHg, sO2 100.0%, FO2Hbe 99.2%. Adequate perfusion using the SAAP circuit must be achieved in order to attain ROSC. Manning and colleagues demonstrated that a rate of 10 mL/kg of body weight per min is the optimum flow rate to raise coronary perfusion pressure (CPP) and result in ROSC6. It was also demonstrated that higher flow rates resulting in too much perfusate may result in cardiac overload or pulmonary edema3. The circuit pump has to have enough power to deliver the perfusate not only through the 3/8-inch ID tubing but also through a relatively long and narrow lumen of the SAAP catheter which measures 81 cm and has a 10 Fr internal diameter. We have found that the centrifugal pump alone is not able to deliver adequate flow rates in this situation, thus a peristaltic pump has been included in the circuit. Adding intra-aortic epinephrine to the SAAP has been shown to lead to more rapid ROSC2. This is best done immediately prior to SAAP perfusion and can be achieved by injecting a small dose (0.2- 0.5 mg) via a three-way stopcock attached to the aortic lumen of the SAAP catheter (Figure 2) and flushed with the SAAP perfusion. The SAAP catheter itself is a crucial element of the whole apparatus. However, no commercially available FDA approved catheter exists for performing SAAP. We have used a prototype catheter which has an internal diameter of 10 Fr, and 13 Fr external diameter, contains a compliant balloon near the tip which can hold up to 50 mL of volume. The catheter has two ports: a balloon port and an arterial lumen port, both of which can accommodate standard IV tubing connectors.
The limitations of this system include the need to calibrate the equipment for blood. Most of commercial non-clinical grade pumps are calibrated to water and given that blood has a different viscosity this may result in different hemodynamic behavior. The equipment described in building the SAAP circuit is not sterile, this should be considered when contemplating survival studies. A porcine model has been extensively used in trauma and cardiovascular research due to anatomic and physiologic compatibility of swine and humans. However, some differences do exist that may influence the outcomes and should be considered when applying this model. The circuit consists of 3/8-inch ID (internal diameter) tubing, which does not have surface coating; however, addition of unfractionated heparin to the collection reservoir combined with recirculation of blood within the circuit seems to mitigate the risk of thrombosis. Although trauma patients can present with coagulopathy7, systemic heparinization is contraindicated, anti-thrombotic measures such as tubing coating should be considered for clinical grade application. For non-survival animal research however, the model can be safely applied. The need to heparinize for systemic heparinization in order to prevent the circuit thrombosis may pose a significant limitation depending on the study protocol. The possible effects of systemic heparinization on myocardial and neurological injury have not studied and cannot be excluded as confounding factors and should be taken under consideration when designing studies with the use of the described circuit. An alternative could be using heparin bonded tubing. Moreover, systemic anticoagulation used in the model is not formally confirmed by laboratory tests and is verified only by lack of thrombosis observed in the circuit. The exsanguination model applied is one of rapid exsanguination. The speed and rate of exsanguination may have an impact on the on the survival and metabolic as well as physiological markers following successful resuscitation with SAAP. This should be considered when designing studies based on exsanguination models. Further studies should be undertaken to fully elucidate the impact of exsanguination rate and speed on the ability to obtain and maintain good ROSC following resuscitation.
Currently there are no commercially available circuits that are specifically developed to deliver SAAP. A circuit for SAAP must have the following capabilities: ability to store perfusate (blood) without the risk of thrombosis, adequate oxygenation and efficient delivery of perfusate at a specific flow rate. Preventing coagulation is paramount when considering storing blood in a circuit even if it is of a short duration. Whilst commercially available circuit tubing for clinical use has surface coating to prevent platelet adhesion, this type of tubing is designed to be single use and the antithrombotic coating usually becomes ineffective after about 14 days8. Modified circuits were constructed by Barnard, Hoops and colleagues9,10 who utilized a 3 L reservoir, a peristaltic pump, and a Cardiohelp system, with an HLS-7 circuit – typically available for ECMO. Clinical grade circuit elements, however, are expensive and cost in the range of $30,000.00 and $130,000.00 depending on the manufacturer (Medtronic and Gentinge, personal communication, April 14, 2020). Moreover, as these circuit elements are not specifically designed for SAAP, they still require some modification, and additional equipment such as peristaltic pump. The cost of the circuit here was significantly lower at $13,500.00, with the pumps constituting the majority of it.
The current report presents a porcine model of traumatic PEA exsanguination cardiac arrest and a specially built circuit which can effectively deliver SAAP (Figure 1). The model of controlled exsanguination used in this study utilized a logarithmic trend with initially rapid rates of bleeding which gradually slow down to a plateau. This model was developed to provide a reproducible hemorrhage which reflects decreasing bleeding rate due to hypotension, and the resulting physiological sequalae. The algorithm can be used to produce logarithmic rates of bleeding to cardiac arrest of varying length depending on the study protocol. SAAP resuscitation has demonstrated promising outcomes in animal models of traumatic and non-traumatic cardiac arrest2,3,4,11. This technique presents an attractive adjunct in the era of emerging endovascular resuscitation. Further studies are needed to further explore its effectiveness in resuscitation of patients with cardiac arrest before the technique is adopted into clinical realm. Currently described SAAP circuit is a cost effective and reliable method of studying the technique before a dedicated circuit is available. The logarithmic, non-linear hemorrhage model presents a reproducible and reliable model for studying resuscitation in traumatic PEA.
The critical steps within the protocol described are relatively rapid logarithmic exsanguination12 and SAAP resuscitation. Exsanguination is performed logarithmically using a programmable pump where the rate of bleeding calculated is based on the animal’s weight decreases every minute. Although this can be done manually, a computer-controlled design allows a precise and smoother pattern. To assure adequate flow rates, exsanguination should be performed using circuit tubing and arterial access cannula of sufficient caliber. SAAP technique requires a concentrated effort of multiple rapid tasks performed in succession, starting with ensuring the circuit is adequately prepared to rapid manual infusion of the AV closure bolus to smooth and quick transition to SAAP perfusion. Contrast added to the SAAP balloon and the AV closure bolus can aid in visualization of the initial perfusion if fluoroscopy is available. However, it is not compulsory and can be omitted depending on the study protocol.
JJ Morrison is a clinical advisory board member of Prytime Medical Inc. All other authors have nothing to disclose.
The views expressed in this article are those of the author(s) and do not reflect the official policy of the Department of Army/Navy/Air Force, Department of Defense, or U.S. Government.
Funding for this study was received by University of Maryland, School of Medicine.
Name | Company | Catalog Number | Comments |
3/8” ID tubing | Saint-Gobain | E-3603 | This tubing is used throughout the circuit. |
1/4" Tubing | Tygon | E-3603 | 2" segment for a connector between Exsanguination tubing and ECMO cannula |
2-way stopcocks | Harvard Apparatus | 72-2650 | standard stopcock |
3-way | Harvard Apparatus | 72-2658 | Standard stopcock |
Barbed Connectors | Harvard Apparatus | 72-1587 | Y connectors |
Barbed Connectors | Harvard Apparatus | 72-1575 | Straight connectors |
Blood Reservoir | LivaNova | 50715 | This is sold together with the oxygenator |
Cable ties | Commercial Electric | GT-200ST | Standard cable ties. |
Centrifugal pump BVP-Z | ISMATEC | ISM 446 | Centrifugal Pump used for recirculation of blood |
Controlled Peristaltic Dispensing Pump | New Era Pump Systems | NE-9000B | Peristaltic pump for Exsanguination |
ECMO Cannula | Medtronic | 96570-015 | Exsanguination cannula |
Gas tubing | AirLife | 1302 | Standard oxygen tubing |
Oxygen source | AirGas | OX USP300 | Standard oxygen tank with flowmeter |
Oxygenator | LivaNova | 50715 | This is sold together with the reservoir |
Peristaltic pump 1 MCP | ISMATEC | ISM 405 | SAAP peristaltic pump |
SAAP catheter | n/a | n/a | Proprietary catheter designed by Dr. Manning |
Venous catheter | Teleflex | CDC-29903-1A | 9 French single lumen catheter |
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