Elsevier

World Neurosurgery

Volume 84, Issue 2, August 2015, Pages 438-445
World Neurosurgery

Original Article
Percutaneous Needle Puncture Technique to Create a Rabbit Model with Traumatic Degenerative Disk Disease

https://doi.org/10.1016/j.wneu.2015.03.066Get rights and content

Background

Several studies have been conducted to identify the pathogenesis of and manage disk degeneration. To further this research, reliable animal models of disk degeneration are required. In the present study, a percutaneous technique is used to create a rabbit model of degenerative disk disease, and the reproducibility and efficacy of this technique is reported.

Methods

Ten mature male New Zealand white rabbits were included in the present study. The intervertebral disk was injured by a percutaneous technique at the L2-L3, L3-L4, and L4-L5 levels. The center of the disk was identified by C-arm guidance. A 19-gauge spinal needle with a 10-mL syringe was inserted into the center of the disk, and negative pressure was applied. Radiographs including magnetic resonance imaging (MRI) with T2 and lateral x-rays were collected at 1, 4, 9, 15, and 20 weeks. Degeneration was examined using histology at 24 weeks.

Results

Narrowed disk height was not observed until 4 weeks after injury, and a significant change was observed at 9 weeks compared with the control L1-L2 level (P < 0.05). MRI revealed disk degeneration beginning at 9 weeks and full degeneration at 15 weeks. Injured intervertebral disks had higher degeneration, seen using MRI, than uninjured control disks. Disk degeneration was confirmed in all injured levels by histologic examination. Cortical osteophyte formation was not found.

Conclusions

Our percutaneous technique provides a suitable rabbit model of degenerative disk disease to test the safety and efficacy of treatments for disk degeneration, such as stem cell transplantation.

Introduction

Intervertebral disk degeneration (IDD) accounts for >20% of individuals with low back pain. The pathogenesis of IDD is not fully understood, but aging is a major contributing factor. Thus, due to the aging population, the prevalence of degenerative spinal diseases, including IDD, is expected to increase.

IDD is treated mainly with surgical methods, although conservative treatments, including physical therapy, medication, and nerve blocks, are also used. Recent developments in biology, technology, and tissue engineering have enhanced the understanding of the pathogenesis and improved the management of disk degeneration. Much research is now focused on treating the disk itself 4, 23, and many groups of investigators are developing animal models of IDD.

The first animal model of IDD was introduced in 1948 9, 16, using a scalpel to stab the disk during surgery, resulting in disk degeneration. Since that time various methods have been explored, and certain key factors were proposed to produce the optimal animal model of IDD. The experimental animal should be easily obtained and inexpensive to manage. The disk needs to be easy to approach surgically and be of adequate size. Disk degeneration should occur reliably and quickly (10). Ideally, a disk degeneration model should be clinically applicable, progressive and reproducible, use a small animal, and be detectable by conventional radiography within 1 month (16).

In the present study, we present a percutaneous technique to induce degenerative disk disease in rabbits, and report the reproducibility and efficacy of this technique.

Section snippets

General Overview

Ten New Zealand white rabbits (2.5 kg body weight) were used with Institutional Animal Care Use Committee (number: HY-IACUC-10-029) approval. Anesthesia was induced with an intramuscular injection of 27.78 mg/kg tiletamine/zolazepam (Zoletil; Virbac Laboratories, Carros, France) and 0.647 mg/kg 2% xylazine hydrochloride (Rompun; Bayer Korea, Seoul, Korea). Percutaneous surgical intervention was performed at the intervertebral disks (IVD) at levels L2-L3, L3-L4, and L4-L5. The L1-L2 level was

Findings Related to the Surgical Intervention

Two of the 10 rabbits suffered complications after surgery. These complications were death (1 rabbit) and weakness of the right hind limb (1 rabbit). Data from the rabbit with motor weakness were included because the rabbit survived until the final evaluation, without abnormal growth when compared with the other rabbits.

Ratio of the IVD Change

Plain x-ray images did not show definite cortical osteophyte formation until the final evaluation. Although the IVD height did not decrease until 4 weeks on plain radiographs, a

Anatomic and Physiologic Considerations of the Rabbit IDD Model

Rabbits are quadrupeds with 7 lumbar vertebrae. Unlike bipeds, degenerative changes are not concentrated at lower lumbar disk levels, but can occur equally at all lumbar spine levels. Therefore, different levels within the same animal were used as control and injured disks.

The L5-L6 and L6-7 disk spaces can only be exposed by open surgery from an anterior approach (10). These levels are not easy to access by percutaneous needle puncture due to the iliac bone covering the posterolateral aspect

Conclusions

The percutaneous needle puncture technique is useful because it is an easy technique, is noninvasive, uses a low dose of anesthetics, has a low risk of death, and induces gradual disk degeneration. Therefore, this technique is useful to produce degeneration in rabbits and to research the management of intervertebral disk disease.

Acknowledgments

The English grammar was revised by eWorldEditing, Inc. and supported by Hanyang University. The English grammar was additionally revised by Bioedit.

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    Conflict of interest statement: This work was supported by the research fund of Hanyang University (HY-2012-G).

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