中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (11): 1675-1679.doi: 10.12307/2022.350

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

动力失衡模型兔颈椎间盘病理改变及终板软骨细胞的迁移凋亡规律

于  栋，刘  侃，时宗庭，杨骁侠，刘恒平，张清烽   

  1. 北京中医药大学第三附属医院脊柱科，北京市   100029
  • 收稿日期:2020-11-18 修回日期:2021-01-16 接受日期:2021-09-11 出版日期:2022-04-18 发布日期:2021-12-11
  • 通讯作者: 刘侃，博士，主治医师，北京中医药大学第三附属医院脊柱科，北京市 100029
  • 作者简介:于栋，男，1976年生，吉林省磐石市人，汉族，2006年中国中医科学院毕业，博士，教授，硕士生导师，主要从事脊柱退行性疾病的临床及基础研究。
  • 基金资助:
    北京中医药大学第三附属医院院级孵化项目(2019kyfh-09)，项目负责人：于栋

Pathological changes of the cervical intervertebral discs and rules of migration and apoptosis in endplate chondrocytes in a rabbit model of dynamic disequilibrium

Yu Dong, Liu Kan, Shi Zongting, Yang Xiaoxia, Liu Hengping, Zhang Qingfeng   

  1. Department of Spine, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2020-11-18 Revised:2021-01-16 Accepted:2021-09-11 Online:2022-04-18 Published:2021-12-11
  • Contact: Liu Kan, MD, Attending physician, Department of Spine, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
  • About author:Yu Dong, MD, Professor, Master’s supervisor, Department of Spine, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China
  • Supported by:
    the Incubation Project of the Third Affiliated Hospital of Beijing University of Chinese Medicine, No. 2019kyfh-09 (to YD)

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摘要：

文题释义：
颈椎的静力平衡和动力平衡：前者又称内源性稳定，包括椎体、附件、椎间盘及相连的韧带；后者又称外源性稳定，包括肌肉的调节和控制，这是颈椎运动的原始动力。
细胞凋亡：是多细胞有机体为调控机体发育、维护内环境稳定，由基因控制的细胞主动死亡过程。细胞凋亡与细胞产生一样是机体自然的生理过程,两者并存，且保持动态平衡，如果细胞凋亡过多或过少，破坏这种平衡，就会导致疾病的发生。

背景：颈肌在维持椎体力学平衡以及颈椎病致病的每一阶段、每一环节都发挥着重要的作用，而且是临床多种颈部症状的主导病因，颈椎力学失平衡及颈椎间盘的退变可看作是以颈肌为主的软组织病变的结果。
目的：观察颈前肌短缩痉挛所致颈椎动力失衡模型兔颈椎间盘终板软骨细胞的病理变化，以及异常应力下椎间盘各结构细胞的凋亡规律。
方法：健康成年新西兰白兔28只，随机分成模型组(n=14)、假手术组(n=14)。模型组于甲状软骨外下方约1 cm处将双侧胸锁乳突肌用医用硅胶硬管垫起致其短缩，建立颈椎动力失衡模型；假手术组仅暴露双侧胸锁乳突肌分离其中部后，直接逐层缝合。于造模后2个月同一时间取材。颈椎间盘及终板软骨组织切片苏木精-伊红染色，光镜下观察颈椎间盘终板软骨细胞的迁移变化情况，运用TUNEL法检测颈间盘终板软骨、纤维环中细胞的凋亡情况。实验方案经北京中医药大学第三附属医院动物实验伦理委员会批准。
结果与结论：①模型组颈椎间盘发生明显的病理学变化，而假手术组未见明显改变；模型组颈椎间盘终板关节软骨区与纤维环区界限明显，未发现明显细胞迁移现象，但椎间盘终板生长软骨区细胞已明显向关节软骨区迁移；模型组椎间盘关节软骨区、生长软骨区出现细胞过度凋亡，其凋亡指数显著高于假手术组(P < 0.01)，纤维环区细胞亦出现一定程度的凋亡，其凋亡指数较假手术组有明显增加(P < 0.05)；②结果表明，颈前肌短缩痉挛所致动力失衡(颈椎肌性痉挛所致的异常应力)，短期内未发现终板软骨与纤维环之间的细胞迁移，但早期即可出现生长软骨层细胞明显向关节软骨层迁移，且椎间盘的终板软骨、纤维环区发生显著的细胞凋亡现象，可为研究椎间盘早期退变的病因机制提供实验证据。

https://orcid.org/0000-0003-3589-9629 (于栋) 

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

关键词: 肌痉挛, 椎间盘退变, 颈椎, 终板, 软骨细胞, 凋亡, 兔

Abstract: BACKGROUND: Cervical muscles play an important role in maintaining the mechanical balance of the vertebral body and the pathogenesis of cervical spondylosis at each stage and link. They are also the leading cause of various clinical cervical symptoms, and mechanical imbalance of the cervical spine and degeneration of the cervical discs can be regarded as the results of soft tissue lesions dominated by cervical muscles.
OBJECTIVE: To observe the pathological changes of endplate chondrocytes in cervical intervertebral discs in a rabbit model of dynamic disequilibrium due to anterior cervical muscle spasm and to study the rules of apoptosis in intervertebral disc cells under abnormal stress.
METHODS: Twenty-eight healthy adult New Zealand rabbits were divided randomly into model group and sham operation group (n=14 per group). The bilateral sternocleidomastoid muscles of the rabbits in the model group were shortened by a medical pipe approximately 1 cm below the outer and lower thyroid cartilage to establish an animal model of cervical dynamic disequilibrium. In the sham operation group, bilateral sternocleidomastoid muscles were only exposed followed by layer-by-layer suture. Endplate cartilage tissue of the cervical intervertebral discs was simultaneously taken from each group at 2 months after modeling, and morphological changes were observed using hematoxylin-eosin staining. The migration of endplate chondrocytes in the cervical intervertebral discs were observed under light microscope. TUNEL method was used to detect the apoptosis of cells in the endplate cartilage and annulus fibrosus. The study protocol was approved by the Animal Experimental Ethics Committee of the Third Affiliated Hospital of Beijing University of Chinese Medicine. 
RESULTS AND CONCLUSION: Obvious pathological changes occurred in the cervical intervertebral disc in the model group, but not in the sham operation group. There was a clear boundary between the articular cartilage area and annulus fibrosus area of the endplate in the model group. No obvious migration was found in the model group, but the cells in the growth cartilage area of the endplate obviously migrated into the articular cartilage area. The model group showed excessive apoptosis in the articular cartilage area and growth cartilage area, and the apoptotic index was significantly higher than that of the sham operation group (P < 0.01). Apoptosis was also found in the annulus fibrosus area, and the apoptotic index was extremely significantly higher than that in the sham operation group (P < 0.05). To conclude, when a dynamic disequilibrium is caused by anterior cervical muscle spasm (abnormal stress caused by cervical muscular spasm), no obvious migration of cells between endplate cartilage and annulus fibrosus occurs in the short term. However, the cells in the growth cartilage area can migrate into the articular cartilage area at an early stage, and significant apoptosis occurs in the endplate cartilage and annulus fibrosus area of the intervertebral disc, which can provide experimental evidence for studying the etiological mechanism of early degeneration of the intervertebral disc.

Key words: muscle spasm, intervertebral disc degeneration, cervical intervertebral disc, endplate, chondrocyte, apoptosis, rabbit

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