An Image Guided Transapical Mitral Valve Leaflet Puncture Model of Controlled Volume Overload from Mitral Regurgitation in the Rat

Summary

A rodent model of left heart volume overload from mitral regurgitation is reported. Mitral regurgitation of controlled severity is induced by advancing a needle of defined dimensions into the anterior leaflet of the mitral valve, in a beating heart, with ultrasound guidance. 

Abstract

Mitral regurgitation (MR) is a widely prevalent heart valve lesion, which causes cardiac remodeling and leads to congestive heart failure. Though the risks of uncorrected MR and its poor prognosis are known, the longitudinal changes in cardiac function, structure and remodeling are incompletely understood. This knowledge gap has limited our understanding of the optimal timing for MR correction, and the benefit that early versus late MR correction may have on the left ventricle. To investigate the molecular mechanisms that underlie left ventricular remodeling in the setting of MR, animal models are necessary. Traditionally, the aorto-caval fistula model has been used to induce volume overload, which differs from clinically relevant lesions such as MR. MR represents a low-pressure volume overload hemodynamic stressor, which requires animal models that mimic this condition. Herein, we describe a rodent model of severe MR in which the anterior leaflet of the rat mitral valve is perforated with a 23G needle, in a beating heart, with echocardiographic image guidance. The severity of MR is assessed and confirmed with echocardiography, and the reproducibility of the model is reported.  

Introduction

Mitral regurgitation (MR) is a common heart valve lesion, diagnosed in 1.7% of the general US population and in 9% of the elderly population greater than 65 years of age¹. In this heart valve lesion, improper closure of the mitral valve leaflets in systole, causes regurgitation of blood from the left ventricle into the left atrium. MR can occur due to various etiologies; however, primary lesions of the mitral valve (primary MR) are diagnosed and treated more frequently compared to secondary MR². Isolated primary MR is often a result of myxomatous degeneration of the mitral valve, resulting in elongation of the leaflets o....

Protocol

Procedures were approved by the Animal Care and Use Program at Emory University under the protocol number EM63Rr, approval date 06/06/2017.

1. Pre-surgical preparation

1. Steam sterilize surgical instruments prior to the procedure.
2. On the procedure day, transfer rats from housing to surgery, and weigh them.
3. Draw pre-operative and post-operative drugs according to the weight: two doses of Carprofen (2.5 mg/kg each), one dose of Gentamycin (6 mg/kg), and one dose of Buprenorphine (0.02 mg/kg).
4. Ensure adequate volume of isoflurane in the gas mixer, and oxygen in the tanks are available for the surgery. On....

Results

Feasibility and reproducibility
The proposed MR model is highly reproducible, with a well defined hole in the mitral leaflet achieved in 100% of the rats used in this study. Figure 6A depicts the direction of the needle as it is inserted into the mitral valve. Figure 6B depicts a hole in the mitral valve leaflet from a representative rat explanted at 2 weeks after the proce.......

Discussion

A reproducible rodent model of severe MR with good survival (93.75% survival after surgery) and without significant post-operative complications is reported. Real-time imaging with transesophageal echocardiography and introduction of a needle into the beating heart to puncture the mitral leaflet are feasible and can be taught. Severe MR was produced with the 23 G needle size in this study, which can be varied as desired using a smaller or larger needle. MR induced in this model creates a low-pressure volume overload on t.......

Materials

Name	Company	Catalog Number	Comments
23G needle	Mckesson	16-N231
25G needle, 5/8 inch	McKesson	1031797
4-0 vicryl	Ethicon	J496H
6-0 prolene	Ethicon	8307H
70% ethanol	McKesson	350600
ACE Light Source	Schott	A20500
ACUSON AcuNav Ultrasound probe	Biosense Webster	10135936	8Fr Intracardiac echo probe
ACUSON PRIME Ultrasound System	Siemens	SC2000
Betadine	McKesson	1073829
Blunted microdissecting scissors	Roboz	RS5990
Buprenorphine	Patterson Veterinary	99628
Carprofen	Patterson Veterinary	7847425
Chest tube (16G angiocath)	Terumo	SR-OX1651CA
Disposable Surgical drapes	Med-Vet	SMS40
Electric Razor	Oster	78400-XXX
Gentamycin	Patterson Veterinary	78057791
Heat lamp with table clamp	Braintree Scientific	HL-1 120V
Hemostatic forceps, curved	Roboz	RS7341
Hemostatic forceps, straight	Roboz	RS7110
Induction chamber	Braintree Scientific	EZ-1785
Injection Plug, Cap, Luer Lock	Exel	26539
Isoflurane	Patterson Veterinary	6679401725
Mechanical ventilator	Harvard Apparatus	Inspira ASV
Microdissecting forceps	Roboz	RS5135
Microdissecting spring scissors	Roboz	RS5603
Needle holder	Roboz	RS6417
No. 15 surgical blade	McKesson	1642
Non-woven sponges	McKesson	446036
Otoscope	Welch Allyn	23862
Oxygen	Airgas Healthcare	UN1072
Pulse Oximeter	Nonin Medical	2500A VET
Retractor, Blunt 4x4	Roboz	RS6524
Rodent Surgical Monitor	Indus Instruments	113970	The integrated platform allows for monitoring of vital signs and surgical warming
Scale	Salter Brecknell	LPS 150
Scalpel Handle	Roboz	RS9843
Silk suture 3-0	McKesson	220263
Small Animal Anesthesia System	Ohio Medical	AKDL03882
Sterile saline (0.9%)	Baxter	281322
Sugical Mask	McKesson	188696
Surgical cap	McKesson	852952
Surgical gloves	McKesson	854486
Syringe 10mL	McKesson	1031801
Syringe 1mL	McKesson	1031817
Ultra-high frequency probe	Fujifilm Visualsonics	MS250
Ultrasound gel	McKesson	150690
VEVO Ultrasound System	Fujifilm Visualsonics	VEVO 2100