graft preparation for loading in ex vivo normothermic machine perfusion systems

<meta charset="utf8"></head><body>Evaluación cardíaca mejorada en perfusión ex vivo con carga ventricular izquierda

Orthotopic heart transplantation is the current gold standard of care for end-stage heart failure1. Unfortunately, the field is significantly limited by a severe donor shortage crisis, resulting in only 2,000 heart transplants being performed each year when over 20,000 people would benefit from the lifesaving procedure2. This organ shortage is expected to worsen as the prevalence of heart failure in the United States alone is projected to surpass 8 million individuals by 20303. Steady increases in waitlist survival times - as a result of improved medical management, advances in mechanical circulatory support, and amendments to the UNOS allocation policy - have resulted in a further increase in the number of patients in need of transplantation at any given moment4,5.
Ex vivo machine perfusion or normothermic machine perfusion (NMP) is a preservation modality that has facilitated the expansion of the supply pool by allowing the use of organs donated after circulatory death (DCD) while achieving some extension of preservation times5,6,7,8. Unlike static cold storage, the current gold standard for preservation, NMP maintains organs in a metabolically active state, which creates the opportunity for real-time monitoring and graft assessment, becoming the standard preservation method for DCD grafts8,9. However, NMP devices currently used clinically are restricted to the Langendorff perfusion mode, which lacks quantitative metrics to predict transplantation outcomes and is unable to capture functional parameters6. For instance, lactate accumulation during Langendorff perfusion has been denoted as the best metabolic predictor of post-transplantation outcomes and is currently used in the clinical setting as a proxy for cardiac graft health10. However, even as the best assessment biomarker, it fails to reliably anticipate the need for mechanical circulatory support post-transplantation11,12. Similarly, the predictive capabilities of commonly utilized hemodynamic parameters (i.e., aortic pressure and coronary blood flow) are largely limited by the retrograde nature of current clinically used configurations for heart machine perfusion9.
The development of assessment protocols for accurate and precise determination of cardiac graft health during NMP would have an immense impact in the field beyond improving post-transplantation outcomes. Objective predictive tools would enable the reliable evaluation and likely utilization of marginal or extended criteria organs (i.e., prolonged warm (&#62; 30 min) and cold ischemia times (&#62; 6 h), increased donor age (&#62; 55), other comorbidities, etc.) from both DCD and brain death donors (DBD) that are currently rejected for transplantation due to the stringent selection criteria13. By enabling the use of marginal hearts, NMP could facilitate an increase in the organ supply as it is estimated that successful transplantation of half the hearts that currently go unused would be sufficient to eliminate the heart waitlist within 2-3 years14. Hemodynamic measurements obtained from left ventricular loading during NMP have garnered significant attention within the field due to their potential as objective assessment parameters. Previous studies have demonstrated that these parameters, such as left ventricular pulse pressure, contractility, and relaxation, are more indicative of cardiac graft function than metabolic trends15,16,17.
In effect, efforts have been dedicated to the development and identification of optimal loading methods to maximize assessment accuracy. Through these efforts, other groups have identified the most relevant mode of aortic perfusion during loading, whereby a stronger correlation between hemodynamic parameters and post-transplantation function was seen when implementing a passive afterload (i.e., no retrograde perfusion to the aorta during loading) when compared with pump-supported afterload (i.e., retrograde perfusion to the aorta during loading)18. This indicates that assisted coronary perfusion likely masks functional deficiencies. Previous studies have successfully incorporated passive afterloads into perfusion setups by implementing systems that mimic the Windkessel effect18,19,20. The Windkessel effect aids in dampening the fluctuation in blood pressure, maintaining continuous blood flow to the tissues and improving coronary perfusion. This protocol achieves the Windkessel-based passive afterload using a modified intravenous (IV) bag enclosed in two spring-loaded plates, where coronary perfusion is exclusively dependent on heart ejection (patent pending).
The use of passive left atrium (LA) pressurization (i.e., gravity-dependent pressurization) during loading, although common practice in small animal heart perfusions, is rarely utilized in the loading of large hearts21,22,23. Instead, the large majority of methods reported in the literature rely on secondary pumps for LA pressurization18,24,25,26,27,28. The pressurization of the LA through a gravity-dependent reservoir, rather than by pump, significantly simplifies the implementation of loading protocols. The use of gravity provides a fixed and constant pressure source, which greatly decreases the need for complicated control systems to achieve and maintain adequate LA pressurization. Moreover, through this pressurization approach, the requirement for a secondary pump is eliminated, facilitating the incorporation of loading capabilities into current Langerdoff setups, as only an extra reservoir is needed. The integration of loading capabilities into clinically utilized machine perfusion systems would amplify the application of cardiac NMP devices by facilitating detailed assessment of cardiac grafts during the preservation period. In effect, maximizing the utility of a system that poses significant financial commitment for patient care due to transportation and device utilization29.
This protocol demonstrates the feasibility of employing both passive afterload and passive LA pressurization during left ventricular loading. Through the validation of passive afterload/LA pressurization as a loading method, this protocol also provides an easy and effective manner of incorporating loading capabilities into established Langendorff perfusion systems. Importantly, it highlights the capability of functional assessment to uncover differences in viable versus failing hearts after extended periods of preservation (&#707;6 h).

<meta charset="utf8"></head><body>Autor destacado: Mejora de la evaluación de la viabilidad del injerto a través de métricas cuantitativas y sistemas de reservorios innovadores

Carga cardíaca mediante presurización pasiva de la aurícula izquierda y poscarga pasiva para la evaluación del injerto

Preparación del injerto para la carga en sistemas de perfusión con máquina normotérmica ex vivo

graft-preparation-for-loading-ex-vivo-normothermic-machine-perfusion

Research

JoVE Journal

Medicine

Graft Preparation for Loading in Ex Vivo Normothermic Machine Perfusion Systems

36 vistas. Para comenzar, tome el corazón de cerdo aislado y corte las ramas del arco aórtico para formar un solo tracto de salida. Inserte la cánula aórtica a través de este tracto y asegúrela con una brida y una sutura de seda 4-0. A continuación, coloque un alambre de estimulación bipolar en la pared posterior del ventrículo derecho.Cree dos suturas de prolene 4-0 por cadena alrededor del perímetro del tracto de la aurícula izquierda. Asegure las su...

Video: Preparación del injerto para la carga en sistemas de perfusión con máquina normotérmica ex vivo

Cardiac Loading using Passive Left Atrial Pressurization and Passive Afterload for Graft Assessment

Ex vivo machine perfusion or normothermic machine perfusion is a preservation method that has gained great importance in the transplantation field. Despite the immense opportunity for assessment due to the beating state of the heart, current clinical practice depends on limited metabolic trends for graft evaluation. Hemodynamic measurements obtained from left ventricular loading have garnered significant attention within the field due to their potential as objective assessment parameters. In effect, this protocol provides an easy and effective manner of incorporating loading capabilities to established Langendorff perfusion systems through the simple addition of an extra reservoir. Furthermore, it demonstrates the feasibility of employing passive left atrial pressurization for loading, an approach that, to our knowledge, has not been previously demonstrated. This approach is complemented by a passive Windkessel base afterload, which acts as a compliance chamber to maximize myocardial perfusion during diastole. Lastly, it highlights the capability of capturing functional metrics during cardiac loading, including left ventricular pulse pressure, contractility, and relaxation, to uncover deficiencies in cardiac graft function after extended periods of preservation times (&#707;6 h).

The protocol describes a porcine ex vivo heart perfusion system in which direct loading of the left ventricle may serve as an assessment technique for graft health while simultaneously providing a holistic evaluation of graft function. A discussion of the system design and possible assessment metrics is also provided.

Ex vivo machine perfusion or normothermic machine perfusion is a preservation method that has gained great importance in the transplantation field. ...