Implantation of Engineered Tissue in the Rat Heart

Summary

Here, we describe a cardiac surgical procedure to implant engineered tissue in the atrioventricular (AV)-groove of an adult Lewis rat.

Abstract

Rodent surgery is often an important component in assessing the utility of engineered tissues. A wide variety of surgical procedures can be performed in common laboratory rats or mice and these quite frequently serve as an intermediate step between bench-top experiments and large animal testing or human trials. Given that rodents provide an established, cost-effective, and physiologically-relevant model system in which to test novel combinations of scaffolding materials and cells, they are particularly well-suited for cardiovascular tissue engineering studies. Presently, we describe an open-heart surgical procedure to implant engineered tissue containing myogenic progenitor cells in the atrioventricular (AV) groove of a rat heart. These implants are intended to create an electrical conduit between the right atrium and right ventricle with the ultimate goal of providing an alternative treatment to conventional pacemaker implantation in pediatric patients with complete heart block[1]. The engineered tissue is implanted in the AV-groove by means of a thoracotomy. For our purposes, Lewis rats are anesthetized and invasively ventilated to maintain positive airway pressure during the sterile surgical procedure. The approach to the heart is performed by a right thoracotomy through an antero-lateral incision at the 5^th intercostal space. The tissue construct is fixed in the AV groove using a single 7-0 Prolene suture and positioned between the right ventricle and atrium at the ventral portion of the heart. The epicardium is partially removed to allow direct contact between the recipient myocardial cells and those contained in the engineered tissue. Following implantation, the chest wall is closed in layers, any pneumothorax is evacuated, and the animal is extubated and treated with analgesic.

Protocol

Part 1: Induction of general anesthesia

1. A 200-250 g Lewis rat is anesthetized with 50 mg/kg Ketamine and 4 mg/kg Xylazine injected with a 30 G needle into the intraperitoneal cavity. The upper abdomen and right chest are clipped to remove hair and the incision site thoroughly cleaned with 70% ethanol and then a povidone-iodine topical antiseptic solution.
2. The animal is intubated endotracheally using a 16 G intravenous catheter connected to a small animal respirator (Harvard Apparatus). The respirator is connected to a vaporizer to supply 0.5 to 1.0% Isoflurane (Baxter) mixed with 100% oxygen to maintain general anesthesia. Respiration rate and t....

Discussion

For a favorable outcome to the surgical procedure, it is critical to properly intubate and invasively ventilate the animal. If the rat is simply sedated and allowed to spontaneously breathe, both lungs will inevitably collapse while the thorax is opened due to common pleura in rodents. Before closing the chest, any pneumothorax needs to be completely eliminated and the lungs fully expanded to reduce atelectasis. Otherwise, weaning from the respirator and recovery will be unsuccessful and the animal will become hypercapni.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
7x4 Thermo Pad 4/box	Equipment	Hood Thermo-Pad, Summerland, BC Canada	MODEL 704R
Inspira ASV Ventilator	Equipment	Harvard Apparatus
Handheld Pulse Oximeter	Equipment	Nellcor	N20PA
Retractor	Equipment	Roboz Surgical Instruments Co.	RS-6514
Isoflurane (Forane)	Reagent	Baxter Internationl Inc.	1001936040
Ketamine HCl	Reagent	Hospira Inc.	RL-0065
Xylazine (AnaSed)	Reagent	Lloyd, Inc.	LB15705A
Buprenorphine HCL Inject	Reagent	Bedford Laboratories	Div-BUP-P00
Bupivacaine 0.25%	Reagent	Hospira Inc.