针刺抽吸髓核法建立椎间盘退变动物模型研究

发布时间：2018-06-20 22:25

  本文选题：椎间盘/病理学 + 模型　； 参考：《福建中医学院》2008年硕士论文

【摘要】： 目的:模拟人后路髓核摘除术,采用后外侧入路经纤维环穿刺抽吸髓核建立兔椎间盘退变动物模型,分析其可行性和优点,并通过影像学和病理学检测分析,再现椎间盘退变过程的客观发展规律,为研究椎间盘退变提供更为理想的动物模型,并为运用组织工程等方法修复重建椎间盘提供实验基础。 方法:日本大耳白兔20只,将每只兔的L1/2和L3/4椎间盘纳入实验组,L2/3椎间盘纳入对照组。术前用速眠新行肌注麻醉,常规术前准备,取左侧卧位,采用右后外侧入路,沿兔L1/2和L3/4右后外侧肌间隙分离,暴露横突和倒“八”字区域后外侧纤维环,直视下用10ml无菌一次性注射器配合21G穿刺针,用无菌塑料套简套住穿刺针,使其尖端保留5mm长度,于纤维环后外侧方平行终板方向刺入,深度控制在5mm,然后注射器回抽约10ml,吸出5.0-8.0mg髓核。逐层缝合关闭切口。动物苏醒后随机分为2、4、8、12周4个模型组,每组5只,分笼常规饲养。术后予以肌注庆大霉素3天预防切口感染。术前及术后2、4、8、12周分别对实验组和对照组椎间盘行CR及MRI检查后,分别将兔子处死,取L1/2、L3/4和L2/3椎间盘进行Masson染色,观察椎间盘组织病理学变化。 结果:(1)影像学分析显示,术后第2周到第12周,X线可见实验组椎间隙变窄,DHI%随造模时间延长而减小,与对照组相比各时间段的差异均有显著性意义(P＜0.05),软骨终板和椎体边缘骨质密度增高。MRI可见实验组椎间盘髓核面积缩小,纤维环面积增大,髓核内MRI T2加权信号降低。髓核信号强度呈逐渐降低趋势,与对照组相比各阶段MRI改良Thompson分级逐渐偏向严重分级,结果差异有显著性意义(P＜0.05)。(2)病理学检测显示,Masson染色可见对照组髓核组织中髓核形态规则,与纤维环界线清晰,髓核细胞较多,纤维环排列整齐致密;实验组髓核裂隙形成,胶原排列紊乱,髓核细胞数量不断减少;随时间增长,类软骨细胞数量减少,髓核组织逐渐纤维化,纤维环逐渐分层紊乱、扭曲、断裂。与对照组相比各时间段Nishimura椎间盘病理分级差异有显著性意义(P＜0.05)。 结论:(1)本实验通过影像学分析和组织病理学检测证实了兔椎间盘退变的基本特征;(2)模拟人后路髓核摘除术,进行后外侧纤维环穿刺抽吸髓核可以诱导兔椎间盘缓慢退变,能再现椎间盘退变病变发展客观规律,而且模型制作简单,重复性好,是研究椎间盘退变病因学、病理生理学等理想的动物模型,而且为研究临床上椎间盘突出症术后复发及检验可靠、有效的治疗椎间盘退变的新方法,例如生长因子疗法、基因疗法、细胞疗法、组织工程等提供实验基础。
[Abstract]:Objective: to establish a rabbit model of degenerative nucleus pulposus by posterolateral approach with posterior approach of nucleus pulposus extraction, and to analyze its feasibility and advantages, and to analyze the feasibility and advantages of the model by imaging and pathological examination. The objective development law of disc degeneration is reproduced, which provides a more ideal animal model for the study of disc degeneration, and provides an experimental basis for the repair and reconstruction of intervertebral disc by means of tissue engineering. Methods: L1 / 2 and L3 / 4 intervertebral discs of each rabbit were included in the control group. Intramuscular anaesthesia was performed with Shumian before operation. Routine preoperative preparation was performed in the left lateral position. The right posterolateral approach was used to separate the right posterolateral muscle space along the L1 / 2 and L3 / 4 spaces of the right posterolateral muscle, and the transverse process and the backward "eight" region were exposed to the posterolateral fibrous ring. 10ml sterile disposable syringe was used in direct view with 21G puncture needle, and the needle was simply covered with sterile plastic sleeve to keep the length of 5mm at the tip of the needle. The needle was punctured in the direction of parallel end plate on the outer side of the fiber ring. The depth was controlled at 5 mm, then the syringe was pumped back about 10 ml and the 5.0-8.0mg nucleus pulposus was sucked out. Close the incision layer by layer. After waking up, the animals were randomly divided into 4 model groups with 5 rats in each group. After operation, gentamicin was injected intramuscularly for 3 days to prevent incision infection. The intervertebral discs of the experimental group and the control group were examined by CR and MRI respectively before operation and at 812 weeks after operation. The rabbits were killed, and the intervertebral discs of L 1 / 2 / 2, L 3 / 4 and L 2 / 3 were taken for Masson staining to observe the histopathological changes of the intervertebral discs. Results the imaging analysis showed that the narrowing of the vertebral space in the experimental group decreased with the prolongation of the time of modeling, and the X ray showed that the narrowing of the vertebral space in the experimental group decreased with the prolongation of the time of modeling at the 2nd week and 12th week after operation. Compared with the control group, the difference was significant (P < 0.05). The density of cartilage endplate and margin of vertebral body was increased. MRI showed that the area of nucleus pulposus decreased, the area of fibrous ring increased, and the signal intensity of T2-weighted MRI in nucleus pulposus decreased in the experimental group. The signal intensity of the nucleus pulposus decreased gradually, and the improved Thompson grading was more severe than that in the control group. Results the difference was significant (P < 0.05) pathological examination showed that the morphology of nucleus pulposus in the control group was regular, the boundary of the nucleus pulposus was clear, the nucleus pulposus cells were more, and the fibrous ring was arranged neatly and compactly, the fissure of nucleus pulposus was formed in the experimental group. With the increase of time, the number of chondroid cells decreased, the tissue of nucleus pulposus became fibrosis, and the fibrous ring was gradually stratified, distorted and broken. Compared with the control group, the pathological grade of Nishimura intervertebral disc was significantly different from that of the control group (P < 0.05). Conclusion (1) the basic characteristic of rabbit disc degeneration was confirmed by imaging analysis and histopathological examination. (2) the posterior approach nucleus pulpotomy can induce the slow degeneration of the intervertebral disc in rabbits, and the nucleus pulposus can be sucked through the posterolateral fibrous ring to induce the slow degeneration of the intervertebral disc. It is an ideal animal model for studying the etiology and pathophysiology of intervertebral disc degeneration, which can reproduce the objective law of the degenerative lesion of intervertebral disc, and the model is simple and reproducible. It also provides experimental basis for the study of clinical recurrence of intervertebral disc herniation and the reliable and effective treatment of disc degeneration, such as growth factor therapy, gene therapy, cell therapy, tissue engineering and so on.
【学位授予单位】：福建中医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R681.53;R-332

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