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Abstract
Introduction
Protocol
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Medicine

Un modello di ratto di fibrillazione ventricolare e Rianimazione dal convenzionale chiuso petto Tecnica

Published: April 26, 2015
doi:
10.3791/52413
, ,
1Resuscitation Institute,Rosalind Franklin University of Medicine and Science

Summary

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

Abstract

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes – but is not limited to – hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

Introduction

Vicino a 360.000 persone negli Stati Uniti 1 e molti altri in tutto il mondo 2 soffrono di un episodio di arresto cardiaco improvviso ogni anno. I tentativi di ripristinare la vita non richiedono solo che l'attività cardiaca essere ristabilita, ma che il danno agli organi vitali essere evitati, ridotti al minimo, o invertiti. Tecniche di rianimazione cardiopolmonare correnti producono un tasso di rianimazione iniziale di circa il 30%; tuttavia, la sopravvivenza alla dimissione ospedaliera è solo il 5% 1. Disfunzione miocardica, disfunzione neurologica, infiammazione sistemica, malattie intercorrenti, o una combinazione di questi si verificano conto post-rianimazione per la gran parte dei pazienti che muoiono a dispetto di cambio iniziale di circolazione. Pertanto, una maggiore comprensione degli approcci fisiopatologia e romanzo rianimazione sottostanti sono urgentemente necessari per aumentare il tasso di rianimazione iniziale e successiva di sopravvivenza con funzione di organo intatto.

Modalità Animalls di arresto cardiaco hanno un ruolo fondamentale nello sviluppo di nuove terapie di rianimazione, fornendo approfondimenti sulla fisiopatologia di arresto cardiaco e rianimazione e offrendo mezzi pratici per concettualizzare e testare nuovi interventi prima che possano essere testati negli esseri umani 3. Il modello di ratto di petto chiuso rianimazione cardiopolmonare (CPR) descritto qui ha svolto un ruolo importante. Il modello è stato sviluppato nel 1988 da Irene von Planta – ricercatore al momento – ed i suoi collaboratori 4 nel laboratorio del professor Max Harry Weil MD, Ph.D. presso l'Università degli Studi di Scienze della Salute (rinominato Rosalind Franklin University of Medicine and Science nel 2004) ed è stato ampiamente utilizzato nel campo della rianimazione prevalentemente dai compagni del professor Weil e dei loro allievi.

Il modello simula un episodio di arresto cardiaco improvviso con rianimazione tentato con tecniche di RCP convenzionale e comprende quindi inductione di fibrillazione ventricolare (VF), fornendo una corrente elettrica al endocardio del ventricolo destro e prestazione di CPR petto chiusa da un dispositivo a pistone ad azionamento pneumatico mentre contemporaneamente fornire ventilazione a pressione positiva con gas arricchita di ossigeno. Cessazione di VF si ottiene consegna transtoracica di scosse elettriche. Il modello di ratto un equilibrio tra modelli sviluppati negli animali di grandi dimensioni (ad esempio, suini) e modelli sviluppati in animali più piccoli (ad esempio, i topi) che consente l'esplorazione di nuovi concetti di ricerca in modo ben standardizzata, riproducibile ed efficiente con l'accesso a un robusto inventario delle misure pertinenti. Il modello è particolarmente utile nelle fasi iniziali della ricerca per esplorare nuovi concetti e esaminare gli effetti confondenti prima realizzazione di studi in modelli animali più grandi che sono più costosi, ma di maggiore impatto traslazionale.

Una ricerca su Medline per tutti gli articoli peer-reviewed la rendicontazione,modello di ratto analoghe sui avendo VF come meccanismo di arresto cardiaco e una qualche forma di rianimazione torace chiuso ha rivelato un totale di 69 ulteriori studi originali utilizzando il modello da quando è stato pubblicato nel 1988 4. Le aree di ricerca comprendono aspetti fisiopatologici della rianimazione 5-17, fattori che influenzano i risultati 18-30, il ruolo di interventi farmacologici esame vasopressori 31-43, agenti tampone 44, agenti inotropi 45, agenti volti a infarto o protezione cerebrale 46-70, e anche gli effetti delle cellule staminali mesenchimali 71-73.

Il modello e il protocollo descritto in questo articolo non è al momento in uso presso l'Istituto di rianimazione. Eppure, ci sono più possibilità di "personalizzare" il modello basato sulle funzionalità disponibili ai singoli ricercatori e gli obiettivi degli studi.

Protocol

NOTA: Il protocollo è stato approvato dalla cura e l'uso degli animali Comitato Istituzionale presso Rosalind Franklin University of Medicine and Science. Tutte le procedure sono in conformità con la guida per la cura e l'uso di animali da laboratorio pubblicato dal Consiglio Nazionale delle Ricerche. 1. Setup sperimentale e Anestesia Eseguire tarature dei vari segnali da catturare utilizzando un sistema di acquisizione dati (pressioni, temperature, cilindrata, …

Representative Results

Il modello di ratto descritto qui è stato recentemente utilizzato per confrontare gli effetti di due inibitori della sarcolemmal sodio-idrogeno scambiatore isoforma 1 (NHE-1) sulla funzione miocardica e emodinamica durante la compressione del torace e post-rianimazione 61. In precedenza era stato riferito che NHE-1-inibitori riducono la riperfusione miocardica limitando citosolico sodio-indotta e sovraccarico di calcio mitocondriale, e in tal modo contribuire a preservare sinistra distensibilità ventricolar…

Discussion

Fasi critiche nel protocollo

Ci sono passaggi critici nel protocollo. Quando padronanza, la preparazione e il protocollo, agire come succintamente descritte di seguito. La preparazione chirurgica è rapida, avanzando cateteri rapidamente attraverso piccole incisioni attivazione spasmo minimo o nullo nave e posizionando le punte del catetere come previsto, seguita da intubazione tracheale successo dopo una singola o pochi tentativo (s); pertanto, completando la preparazione in ≈ 90…

Disclosures

The authors have nothing to disclose.

Acknowledgements

The authors would like to acknowledge Dr. Wanchun Tang MD, MCCM, FCCP, FAHA and Jena Cahoon of the Weil Institute of Critical Care Medicine in Rancho Mirage, CA. for their contributions to the resuscitation protocol outline and for having helped train the rodent surgeon (LL). The preparation of this article was in part supported by a gift in memory of US Navy Retired SKC Robert W. Ply by Ms. Monica Ply for research in heart disease and Parkinson’s disease and by a discretionary fund from the Department of Medicine at Rosalind Franklin University of Medicine and Science.

Materials

Name of Material/ Equipment Company Catalog Number Comments/Description
Sodium pentobarbital Sigma Aldrich P3761 http://www.sigmaaldrich.com/catalog/product/sigma/p3761?lang=en&region=US
Rectal thermistor BIOPAC Systems, INC TSD202A http://www.biopac.com/fast-response-thermistor
Needle electrode biopolar concentric 25 mm TP BIOPAC Systems, INC EL451 http://www.biopac.com/needle-electrode-concentric-25mm
PE25 polyethylene tubing  Solomon Scientific BPE-T25 http://www.solsci.com/products/polyethylene-pe-tubing
26GA female luer stub adapter Access Technologies LSA-26 http://www.norfolkaccess.com/needles.html
Stopcocks with luer connections; 3-way; male lock, non-sterile Cole-Parmer UX-30600-02 http://www.coleparmer.com/Product/Large_bore_3_way
_male_lock_stopcocks
_10_pack_Non_sterile/EW-30600-23
TruWave disposable pressure transducer Edwards Lifesciences PX600I  http://www.edwards.com/products/pressuremonitoring/Pages/truwavemodels.aspx?truwave=1
Type-T thermocouple Physitemp Instruments IT-18 http://www.physitemp.com/products/probesandwire/flexprobes.html
Central venous pediatric catheter  Cook Medical  C-PUM-301J https://www.cookmedical.com/product/-/catalog/display?ds=cc_pum1lp_webds
Abbocath-T subclavian I.V. catheter (14g x 5 1/2") Hospira 453527 http://www.hospira.com/products_and_services/iv_sets/045350427
Novametrix Medical Systems, Infrared CO2 monitor Soma Technology, Inc. 7100 CO2SMO  http://www.somatechnology.com/MedicalProducts/novametrix_respironics_co2smo_
7100.asp
Harvard Model 683 small animal ventilator Harvard Apparatus 555282 http://www.harvardapparatus.com/webapp/wcs/stores/servlet/haisku2_10001_11051_44453_-1_
HAI_ProductDetail_N_37322_37323
Double-flexible tipped wire guides Cook Medical  C-DOC-15-40-0-2 https://www.cookmedical.com/product/-/catalog/display?ds=cc_doc_webds
High accuracy AC LVDT displacement sensor Omega Engineering LD320-25 http://www.omega.com/pptst/LD320.html
HeartStart XL defibrillator/monitor Phillips Medical Systems M4735A http://www.healthcare.philips.com/main/products/resuscitation/products/xl/
Graefe micro dissection forceps 4 inches Roboz  RS-5135 http://shopping.roboz.com/Surgical-Instrument-Online-Shopping?search=RS-5135
Graefe micro dissection forceps 4 inches with teeth Roboz  RS-5157 http://shopping.roboz.com/Surgical-Instrument-Online-Shopping?search=RS-5157
Extra fine micro dissection scissors 4 inches Roboz  RS-5882 http://shopping.roboz.com/micro-scissors-micro-forceps-groups/micro-dissecting-scissors/Micro-Dissecting-Scissors-4-Straight-Sharp-Sharp
Heiss tissue retractor Fine Science Tools  17011-10 http://www.finescience.com/Special-Pages/Products.aspx?ProductId=321&CategoryId=134&
lang=en-US
Crile curve tip hemostats Fine Science Tools  13005-14 http://www.finescience.com/Special-Pages/Products.aspx?ProductId=372
Visistat skin stapler  Teleflex Incorporated 528135 http://www.teleflexsurgicalcatalog.com/weck/products/9936
Braided silk suture, 3-0 Harvard Apparatus 517706 http://www.harvardapparatus.com/webapp/wcs/stores/servlet/haisku2_10001_11051_43051_-1_
HAI_ProductDetail_N_37916_37936
Betadine solution Butler Schein 3660 https://www.henryscheinvet.com/
Sterile saline, 250 ml bags Fisher 50-700-069 http://www.fishersci.com/ecomm/servlet/itemdetail?catnum=50700069&storeId=10652
Heparin sodium injection, USP Fresenius Kabi 504201 http://fkusa-products-catalog.com/files/assets/basic-html/page25.html
Loxicom (meloxicam) Butler Schein 045-321 https://www.henryscheinvet.com/
Thermodilution cardiac output computer for small animals N/A N/A Custom-developed at the Resuscitation Institute using National Instruments hardware and LabVIEW software
Analog-to-digital data acquisition and analysis system N/A N/A Custom-developed at the Resuscitation Institute using National Instruments hardware and LabVIEW software
Pneumatically-driven and electronically controlled piston device for chest compression in small animals N/A N/A Custom-developed at the Weil Institute of Critical Care Medicine
60 Hz alternating current generator N/A N/A Custom-developed at the Weil Institute of Critical Care Medicine
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Keywords: Rat ModelVentricular FibrillationCardiac ResuscitationClosed-chest TechniqueCardiopulmonary ResuscitationVascular CatheterizationTracheal IntubationPositive Pressure VentilationElectrical ShocksHemodynamic MeasurementsMetabolic MeasurementsResuscitation ConceptsTherapeutic InterventionsCardiac Arrest
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Lamoureux, L., Radhakrishnan, J., Gazmuri, R. J. A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique. J. Vis. Exp. (98), e52413, doi:10.3791/52413 (2015).
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