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Wistar大鼠针刺椎间盘退变实验模型

Experimental model of intervertebral disc degeneration by needle puncture in Wistar rats.

作者信息

Issy A C, Castania V, Castania M, Salmon C E G, Nogueira-Barbosa M H, Bel E Del, Defino H L A

机构信息

Departamento de Morfologia, Fisiologia e Patologia Básica, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil.

出版信息

Braz J Med Biol Res. 2013 Mar;46(3):235-44. doi: 10.1590/1414-431x20122429. Epub 2013 Mar 15.

Abstract

Animal models of intervertebral disc degeneration play an important role in clarifying the physiopathological mechanisms and testing novel therapeutic strategies. The objective of the present study is to describe a simple animal model of disc degeneration involving Wistar rats to be used for research studies. Disc degeneration was confirmed and classified by radiography, magnetic resonance and histological evaluation. Adult male Wistar rats were anesthetized and submitted to percutaneous disc puncture with a 20-gauge needle on levels 6-7 and 8-9 of the coccygeal vertebrae. The needle was inserted into the discs guided by fluoroscopy and its tip was positioned crossing the nucleus pulposus up to the contralateral annulus fibrosus, rotated 360° twice, and held for 30 s. To grade the severity of intervertebral disc degeneration, we measured the intervertebral disc height from radiographic images 7 and 30 days after the injury, and the signal intensity T2-weighted magnetic resonance imaging. Histological analysis was performed with hematoxylin-eosin and collagen fiber orientation using picrosirius red staining and polarized light microscopy. Imaging and histological score analyses revealed significant disc degeneration both 7 and 30 days after the lesion, without deaths or systemic complications. Interobserver histological evaluation showed significant agreement. There was a significant positive correlation between histological score and intervertebral disc height 7 and 30 days after the lesion. We conclude that the tail disc puncture method using Wistar rats is a simple, cost-effective and reproducible model for inducing disc degeneration.

摘要

椎间盘退变的动物模型在阐明生理病理机制和测试新的治疗策略方面发挥着重要作用。本研究的目的是描述一种用于研究的、涉及Wistar大鼠的简单椎间盘退变动物模型。通过X线摄影、磁共振成像和组织学评估来确认和分类椎间盘退变。成年雄性Wistar大鼠麻醉后,在尾椎的6-7和8-9节段用20号针头进行经皮椎间盘穿刺。在荧光透视引导下将针头插入椎间盘,使其尖端穿过髓核直至对侧纤维环,旋转360°两次,并保持30秒。为了评估椎间盘退变的严重程度,我们在损伤后7天和30天从X线图像测量椎间盘高度,并进行T2加权磁共振成像信号强度测量。采用苏木精-伊红染色和用苦味酸天狼星红染色及偏振光显微镜观察胶原纤维方向进行组织学分析。影像学和组织学评分分析显示损伤后7天和30天均有明显的椎间盘退变,无死亡或全身并发症。观察者间的组织学评估显示有显著的一致性。损伤后7天和30天,组织学评分与椎间盘高度之间存在显著正相关。我们得出结论,使用Wistar大鼠的尾椎间盘穿刺方法是一种简单、经济有效且可重复的诱导椎间盘退变的模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d97/3854370/20ce4be569a3/1414-431X-bjmbr-46-03-235-gf01.jpg

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