中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (12): 1855-1860.doi: 10.3969/j.issn.2095-4344.2017.12.009

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

建立纤维环穿刺法椎间盘退变模型

王字兴,蔚  芃   

  1. 川北医学院附属医院骨科,四川省南充市  637000
  • 收稿日期:2016-12-17 出版日期:2017-04-28 发布日期:2017-05-16
  • 通讯作者: 蔚芃,硕士,主任医师,教授,硕士生导师,川北医学院附属医院骨科,四川省南充市 637000
  • 作者简介:王字兴,男,1982年生,四川省古蔺县人,2012年川北医学院毕业,硕士,主治医师,主要从事椎间盘退变方面的研究。
  • 基金资助:

    四川省医学会创伤专项课题(2015GK007)

Establishing an intervertebral disc degeneration model using annulus puncture

Wang Zi-xing, Wei Peng   

  1. Department of Orthopaedics, Affiliated Hospital of North Sichuan Medical University, Nanchong 637000, Sichuan Province, China
  • Received:2016-12-17 Online:2017-04-28 Published:2017-05-16
  • Contact: Wei Peng, Master, Chief Physician, Professor, Master’s supervisor, Department of Orthopaedics, Affiliated Hospital of North Sichuan Medical University, Nanchong 637000, Sichuan Province, China
  • About author:Wang Zi-xing, Master, Attending physician, Department of Orthopaedics, Affiliated Hospital of North Sichuan Medical University, Nanchong 637000, Sichuan Province, China
  • Supported by:

    the Special Project of Sichuan Medical Traumatic Association, No. 2015GK007

摘要:

文章快速阅读:

文题释义:
椎间盘退变:椎间盘的组织结构发生退行性改变过程,称为椎间盘退变。各种物理的、化学的因素外,椎间盘随着年龄的增长,其中椎间盘髓核及纤维坏中细胞及细胞外基质改变、水分逐渐丢失,椎间盘生物特性及物理学特性发生改变。是导致颈肩腰腿痛的病理基础。
椎间盘退变动物模型:被认为是研究椎间盘退变的发生机制和检测各种治疗的一种重要途径,建立一种可靠的椎间盘退变动物模型将为研究椎间盘退变发病机制提供有利的条件,同时为修复退变椎间盘的各种治疗探索研究提供良好的实验载体。目前制作椎间盘退变动物模型的方法主要分为自然退变和实验诱导,自然退变主要有高盐饮食制造地中海沙鼠椎间盘模型、截除双前肢建立双后肢建立大鼠椎间盘退变模型以及随年龄增长椎间盘自发性退变改变等。
摘要
背景:
目前虽然纤维环穿刺法构建椎间盘退变模型的相关研究报道较多,但缺乏相对完整的影像、病理学以及细胞外基质改变(蛋白多糖含量)的系统对比性研究。所以经腹膜外入路暴露椎间隙纤维穿刺法造模的影像、病理学以及细胞外基质改变是否与人类椎间盘退变类似需进一步探讨。
目的:验证采用纤维环穿刺法建立椎间盘退变模型的可行性。
方法:将30只新西兰大白兔随机分为2组,每组15只。纤维环穿刺组在经腹膜外入路暴露L1,2,L2,3,L3,4椎间隙后,采用16号针头穿刺;而假手术组只做切开不做穿刺。于造模术后4,8,16周2组均随机抽取5只行MRI检查后,以空气栓塞法处死兔并切取椎间盘髓核组织,苏木精-伊红染色检查组织学变化,Safranin O染色观察髓核蛋白多糖水平改变。
结果与结论:①MRI提示:纤维环穿刺组T2WI相椎间盘信号自造模术后开始降低,且随着术后时间延长T2WI相椎间盘信号降低越明显,于术后4周较假手术组有明显改变,术后16周最明显;假手术组术后各时间段改变不明显;②苏木精-伊红染色显示:纤维环穿刺组造模术后软骨样细胞减少,纤维样细胞增多且所占比例增大,细胞外纤维排列紊乱,于第4周较假手术组有明显改变,于第16周最明显;假手术组术后各时间段改变不明显;③Safranin O染色(髓核蛋白多糖水平):纤维环穿刺组组织着色于造模术后开始变浅,随着术后时间延长着色变浅越明显,于术后4周较假手术组有明显改变,术后16周最明显;④结果提示:纤维环穿刺法建立的椎间盘退变模型影像学表现及病理表现与人类椎间盘退变近似,提示纤维环穿刺法是建立椎间盘退变模型的可靠方法。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0003-3475-5287(王字兴)

关键词: 组织构建, 软骨组织工程, 造模, 纤维环穿刺法, 椎间盘退变, 髓核组织, Safranin O染色, 蛋白多糖, 兔, 组织学变化, 蛋白多糖

Abstract:

BACKGROUND: Annulus puncture method is commonly reported in the establishment of intervertebral disc degeneration (IDD) models, but there is a lack of comparative studies concerning complete MRI, pathologyical and extracellular matrix changes. Thereafter, a further exploration is imperative.
OBJECTIVE: To investigate the feasibility of establishing IDD models using annulus puncture.
METHODS: Thirty New Zealand white rabbits were randomly divided into two groups (n=15 per group). In experimental group, through extraperitoneal approach, the L1-2, L2-3, and L3-4 intervertebral space were exposed, and then stabbed by 16-gauge hypodermic needles; while rabbits in sham operated group were only subjected to making and incision but not to puncture. Five rabbits in each group were randomly selected for MRI examination at 4, 8 and 16 weeks after modeling, and were sacrificed through air embolism to obtain the nucleus pulposus. Histological changes were observed by hematoxylin-eosin staining, and the level of proteoglycan was detected by Safranin O staining.
RESULTS AND CONCLUSION: In the experimental group, T2-weighted signal in the disc was decreased in a time-dependent manner; an obvious reduction was found at 4 weeks after modeling, and reached the peak at 16 weeks. In contrast, the sham operated group showed no significant changes at each time point. Hematoxylin-eosin staining showed that in the experimental group, cartilage-like cells decreased, while fibroblast-like cells in the disc increased in number and proportion, and extracellular fibers were in disorder; all above changes became obvious at 4 weeks after surgery and the most obvious at 16 weeks, but there were no significant changes in the sham operated group. Safranin O staining revealed that the color of the tissues under puncture became lighter, and this variation became overt at 4 weeks after surgery and the most obvious at 16 weeks. In conclusion, annulus puncture is a reliable method to make IDD models.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Intervertebral Disk Degeneration, Models, Animal, Tissue Engineering

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