Construction of a Composite Regenerative Peripheral Nerve Interface in a Rat Model of Nerve Injury

Abstract

Source: Svientek, S. R., et al. Fabrication of the Composite Regenerative Peripheral Nerve Interface (C-RPNI) in the Adult Rat. J. Vis. Exp. (2020).

This video demonstrates the surgical construction of a composite regenerative peripheral nerve interface (C-RPNI) in a rat model of peripheral nerve injury. The procedure involves attaching a transected peroneal nerve to a muscle graft and overlaying it with a dermal skin graft to promote nerve regeneration. Over time, motor and sensory neurons reestablish connections within the grafts, enabling the formation of functional neuromuscular and sensory structures.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

NOTE: Donor rats are allowed free access to food and water prior to skin and muscle donation procedures. Euthanasia is performed under deep anesthesia followed by intracardiac potassium chloride injection with a secondary method of bilateral pneumothorax. Any strain of rat can theoretically be utilized with this experiment; however, our laboratory has achieved consistent results in both male and female Fischer F344 rats (~200-250 g) at two to four months of age. Donor rats must be isogenic to the experimental rats.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
#15 Scalpel	Aspen Surgical, Inc	Ref 371115	Rib-Back Carbon Steel Surgical Blades (#15)
4-0 Chromic Suture	Ethicon	SKU# 1654G	P-3 Reverse Cutting Needle
5-0 Chromic Suture	Ethicon	SKU# 687G	P-3 Reverse Cutting Needle
6-0 Ethilon Suture	Ethicon	SKU# 697G	P-1 Reverse Cutting Needle (Nylon suture)
8-0 Monofilament Suture	AROSurgical	T06A08N14-13	Black polyamide monofilament suture on a threaded tapered needle
Experimental Rats	Envigo	F344-NH-sd	Rats are Fischer F344 Strain
Fluriso (Isofluorane)	VetOne	13985-528-40	Inhalational Anesthetic
Micro Motor High Speed Drill with Stone	Master Mechanic	Model 151369	Handheld rotary tool; kit comes with multiple fine grit stones
Oxygen	Cryogenic Gases	UN1072	Standard medical grade oxygen canisters
Potassium Chloride	APP Pharmaceuticals	63323-965-20	Injectable form, 2 mEq/mL
Povidone Iodine USP	MediChoice	65517-0009-1	10% Topical Solution, can use one bottle for multiple surgical preps
Puralube Vet Opthalmic Ointment	Dechra	17033-211-38	Corneal protective ointment for use during procedure
Rimadyl (Caprofen)	Zoetis, Inc.	NADA# 141-199	Injectable form, 50 mg/mL
Stereo Microscope	Leica	Model M60	User can adjust magnification to their preference
Surgical Instruments	Fine Science Tools	Various	User can choose instruments according to personal preference or from what is currently available in their lab
Triple Antibiotic Ointment	MediChoice	39892-0830-2	Ointment comes in sterile, disposable packets
VaporStick 3	Surgivet	V7015	Anesthesia tower with space for isofluorane and oxygen canister
Webcol Alcohol Prep	Coviden	Ref 6818	Alcohol prep wipes; use a new wipe for each prep