针刺与异常机械应力对大鼠椎间盘纤维环纳米级生物力学特性影响的对比  

Influence of needle puncture versus abnormal mechanical compression on the biological properties of annulus fibrosus: a preliminary study at nano-scale

作　　者：李海涛 梁婷 邵毅杰 陈曦[2] 杨惠林[1] 罗宗平[2] Li Haitao;Liang Ting;Shao Yijie;Chen Xi;Yang Huilin;Luo Zongping(The First Affiliated Hospital of Soochow University,Suzhou 215000,Jiangsu Province,China;Institute of Orthopaedics at Soochow University,Suzhou 215000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院,江苏省苏州市215000 [2]苏州大学骨科研究所,江苏省苏州市215000

出　　处：《中国组织工程研究》2018年第34期5510-5517,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81320108018;31570943;31270995)~~

摘　　要：背景:目前关于椎间盘退变机制的研究仍不透彻,尤其是不同退变机制影响下退变椎间盘纳米级改变的研究几乎处于空白状态。目的:建立一种简便易行的大鼠尾椎椎间盘退变模型,探索针刺及异常机械应力退变机制对纤维环单根胶原纤维纳米级别生物力学特性的影响。方法:取8只骨发育成熟的雄性SD大鼠,手法定位每只大鼠第5-11尾椎间盘位置,其中Co5-Co6椎间盘为正常组;对Co6-Co7椎间盘实施半程穿刺操作(针刺组);对第7-10椎间盘加装自行设计的外固定支架(其中Co8-Co9椎间盘为异常压应力组);Co10-Co11椎间盘为邻近组。4周后,对大鼠尾椎行磁共振矢状位T2加权像扫描,椎间盘组织行苏木精-伊红与番红O-快绿染色,以及椎间盘组织纤维环内外层原子力显微镜观察。结果与结论:(1)异常压应力组、针刺组大鼠尾椎间盘MRI信号明显低于正常组及邻近组(P <0.05),正常组及邻近组尾椎间盘MRI信号无明显差异;(2)组织学观察显示,邻近组尾椎间盘组织表现与正常组相似;针刺组一侧纤维环明显紊乱、增厚且突入髓核区域之内,髓核与纤维环界限紊乱,穿刺一侧髓核内细胞数量明显下降,空泡状结构消失;异常压应力组纤维环髓核区域面积下降,髓核细胞数量减少、聚集,空泡状结构消失,髓核与纤维环界限清晰;(3)原子力显微镜观察显示,各组纤维环外层的胶原纤维弹性模量均高于内层(P <0.05);邻近组、异常压应力组纤维环内、外层的胶原纤维弹性模量与正常组比较无差异;针刺组纤维环内、外层的胶原纤维弹性模量大于正常组、异常压应力组(P <0.01);(4)结果表明与异常压应力诱导机制不同,针刺机制诱导的椎间盘退变纤维环胶原纤维表现出明显僵硬化现象。BACKGROUND:Currently,the specific pathological mechanism of intervertebral disk degeneration,especially the tissue alteration at nano-scale,is still unknown.OBJECTIVE:To establish a rat model of intervertebral disk degeneration in the caudal vertebrae and to explore the biomechanical alteration of collagen fibrils of annulus fibrosus at nano-scale induced by needle puncture and abnormal mechanical loading.METHODS:Eight adult Sprague-Dawley male rats were chosen and the coccygeal vertebral discs from C5-11 were located under anesthesia.The Co5-Co6 segments were set as normal group.The Co6-Co7 segments were subjected to needle puncture(needle-puncture group).The Co7-Co10 segments were drilled percutaneously by four parallel K-wires and the wires were fixed with self-made braces to make the Co8-Co9 segments under a constant compression environment.The Co8-Co9 segments were set as abnormal compression group.The Co10-Co11 segments were set as adjacent group.Sagittal T2-weighted MRI scanning of the rat caudal vertebrae was performed at 4 weeks after treatment.Hematoxylin-eosin and safranin O/Fast green staining of the intervertebral disc was also performed,and the inner and outer layers of the annulus fibrosus were observed using atomic force microscope.RESULTS AND CONCLUSION:(1)The MRI signals in the abnormal compression group and needle-puncture group were significantly lower than those in the normal and adjacent groups(P<0.05),while the MRI signals in normal and adjacent groups showed no significant difference.(2)Histological analysis revealed similarity in the normal and adjacent groups.In the needle-puncture group,the annulus fibrosus on the puncture site arranged disorderly,thickened and bulged inward into the nucleus pulposus,resulting in an unclear boundary between the annulus fibrosus and nucleus pulposus.The number of nucleus pulposus cells on the puncture side was dramatically reduced and vacuole-like structure disappeared.In the abnormal compression group,nucleus pulposus area remarkably decreased,the numbe

关 键 词：纤维环 大鼠椎间盘退变模型 原子力显微镜 针刺与异常压应力 纳米级 椎间盘退化 显微镜检查 原子力 弹性模量 组织工程 

分 类 号：R318[医药卫生—生物医学工程]