cardiopulmonary bypass (cpb) circuit setup for cardiac arrest and resuscitation in pediatric animal model

Ekstrakorporal hjerte-lunge-redning i dyremodell etter hjertestans

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted&#160;worldwide per year1. Though modern advances in cardiothoracic surgery (CTS) and intensive care treatment have improved mortality rates, comorbidities remain extremely common2,3,4,5. Neurodevelopmental abnormalities, including cognitive and motor impairments as well as learning disabilities, are reported in around 25%-50% of these patients6,7,8. Surgery during the first days of life, especially those that require circulatory and cardiac arrest (CA), has been demonstrated to increase morbidity9. Hemodynamic alterations during surgery may have an important effect on the vulnerable developing newborn brain. Experimental models are essential to better understand the origin of these abnormalities and investigate neuroprotective strategies to improve the prognoses of these patients.
The use of animal models to study this population has been widely documented5,10,11,12,13,14. Notably, piglets offer an excellent option, given close approximations in cardiac anatomy (Figure 1), genome, and physiology, as well as their relatively larger size in comparison to other animal models15 (Figure 2). The use of piglet models to study the effects of both cardiopulmonary bypass (CPB) and CA has been previously described. These experimental animal models are useful for studying hemodynamic changes and associated end-tissue organ complications14,16,17,18,19,20. These models were developed to allow researchers to study human conditions in a controlled setting, with flexibility for a variety of experimental conditions. Most studies report the use of central cannulation, a technique that demands advanced surgical skills, requires higher resource utilization, and makes it difficult to ensure long-term survival. Though previous studies have documented the use of piglets in studying CPB12,15, few have proposed the peripheral cannulation technique.
This new peripheral cannulation technique is easier, less aggressive, and more feasible when compared to other published studies19. Moreover, validating this technique in newborns and small animals is novel and should be considered for use by all researchers interested in using an animal model to study CHD and its associated comorbidities. It is particularly appropriate for individuals with access to a laboratory equipped with supplies, resources, and personnel experienced in conducting animal model experiments.
In summary, the main aim of this study is to describe and validate a neonatal piglet model of CPB with CA. The protocol aims to study severe brain damage and other possible complications of CPB surgery in a controlled setting with varying experimental conditions. This method provides a model that is generalizable, reliable, and of high quality, which can be used for a wide variety of experimental protocols.

Forfatter i søkelyset: Simulering av pediatrisk hjertekirurgi ved hjelp av en neonatal smågrismodell

Pediatrisk dyremodell for ekstrakorporal hjerte-lunge-redning etter langvarig sirkulasjonsstans

Mortality rates for children with congenital heart defects have improved significantly in recent years given improvements in cardiac surgery. However, comorbidities remain extremely common, and that's the goal of our lab. We aim to investigate these comorbidities, uncover underlying causes, and develop preventative measures.Right now, we are studying neuro abnormalities that develop after cardiac surgery. This protocol offers a feasible and reproducible animal model of simulating pediatric cardiac surgery. So with that, researchers can use it as a base to test many different experimental conditions and a wide variety of other hypotheses.The peripheral cannulation technique for porcine cardiopulmonary bypass in our model is both practical and cost effective. Furthermore, it does allow a more reliable way to assess postoperative outcomes than the traditional central cannulation technique. The flexibility of our model encourages future work in congenital heart disease and comorbidity studies.For example, our lab is currently planning to expand this model to study real-time monitoring of cerebral parameters such as oxygenation status and cerebral blood flow.

Kardiopulmonal bypass (CPB) kretsoppsett for hjertestans og gjenoppliving i pediatrisk dyremodell

For å begynne, plasser den fire til seks dager gamle bedøvede smågrisen i ryggliggende stilling. Etter å ha utført orotrakeal intubasjon ved bruk av et mansjettrør, auskulterer du lungebasene for å bekrefte passende plassering av endotrakealrøret. Sett opp mekanisk ventilasjon og overvåk kontinuerlig dybden av anestesi under operasjonen gjennom hjertefrekvens, blodtrykk og oksygenmetning.Juster ventilasjons- og sedasjonsparametere etter behov. Sett deretter inn et 3 fransk kateter i lårarterien, etterfulgt av et 4 fransk kateter i venen. Deretter starter du den kardiopulmonale bypass, eller CPB, kretsen som er satt opp ved å forkorte CPB-slangen mens du opprettholder nok avstand mellom dyret og maskinen.Lag en rørbro for å koble membranoksygenatorens utstrømning og pumpeinnstrømning. Når alle tilkoblingspunktene er forseglet, feier du kretsen med 300 mikroliter heparinsaltløsning og 300 milliliter ferskt donorgriseblod, etterfulgt av natriumbikarbonat, heparin og kalsiumglukonat. Når CPB-kretsen er klar, eksponer du den venstre indre halsvenen og setter nålekateteret inn i den.På samme måte eksponerer du høyre halspulsåre for å forberede deg på kanylering. Når et blodglimt er synlig, sett forsiktig en ledetråd inn i karet. Tre en dilator over ledningen og inn i karet.Fjern deretter dilatatoren. Tre sakte en 8 fransk venekanyle omtrent fire centimeter inn i karet, og fjern ledningen, og sørg for at kanylen forblir på plass. Plasser deretter en 6 fransk pediatrisk arteriell kanyle i høyre halspulsåre som tidligere vist med dilatasjon.Til slutt administreres en intravenøs heparinbolus gjennom den nyplasserte arterielle kanylen. Etter å ha oppnådd tilgang, bruk 3-0 absorberbare suturer for å feste begge kanylene til dyret, og tape for å forhindre utilsiktet fjerning. Koble kanylene til CPB-kretsen, og sørg for at heparinsaltvann tilsettes tilkoblingspunktene for å forhindre luft i kretsen.Sett startstrømmen til 80 til 85 milliliter per kilo per minutt. For å indusere hjertestans, administrer ni milliekvivalenter kaliumklorid. Når hjertet er stoppet, isoler dyret fra kretsen.Hold deretter CPB-kretsstrømmen til å sirkulere ved 1 500 RPM. Etter å ha oppnådd passende hjertestanstilstander, start ekstrakorporal hjerte-lungeredning ved å koble smågrisen til CPB-kretsen igjen. Administrer tre milliliter kalsiumglukonat og seks milliliter natriumbikarbonat gjennom perifer arteriell tilgang, og tilsett doser etter behov.Karkanyleringen og CPB med hjertestans oppnådde en suksessrate på 92 % i eksperimentene. MR viste klare tegn på tidlig hjerneskade i CPB med hjertestansdyr sammenlignet med kontroll. Den akutte diffusjonstensoravbildningsmodaliteten identifiserte bedre tegn på infarkt enn andre konvensjonelle MR-sekvenser.

Research

JoVE Journal

Medicine

Cardiopulmonary Bypass (CPB) Circuit Setup for Cardiac Arrest and Resuscitation in Pediatric Animal Model

Pediatric Animal Model of Extracorporeal Cardiopulmonary Resuscitation After Prolonged Circulatory Arrest

Congenital heart disease (CHD) is the most prevalent congenital malformation, with about one million births impacted worldwide per year. Comprehensive ...