新西兰大白兔椎间盘退变动物模型的比较研究

发布时间：2018-07-03 11:54

本文选题：经皮纤维环穿刺 + 椎间盘退变　；参考：《第四军医大学》2011年硕士论文

【摘要】：理想的椎间盘退变动物模型是椎间盘组织工程研究的必要条件,但目前尚缺乏公认的模型制备方法,探索一种动物创伤性小、造模操作简单的模型制备方法已成为目前研究的当务之急。实验通过生物化学、影像学等检测手段比较C形臂辅助下经皮纤维环穿刺法、右后外侧腹膜外入路纤维环穿刺法及经右后外侧腹膜外入路髓核抽吸法制备兔椎间盘退变动物模型优缺点,评估C形臂辅助下经皮纤维环穿刺法制备椎间盘退变动物模型的可行性。本实验分为两部分,第一部分为椎间盘退行性改变动物模型的建立,第二部分为椎间盘退行性改变动物模型的检测评估。实验选用清洁级新西兰大白兔27只,将实验动物随机分入C形臂辅助下经皮纤维环穿刺组、右后外侧腹膜外入路纤维环穿刺组及右后外侧腹膜外入路髓核抽吸组,每组9只动物。选定每只新西兰大白兔L2/3和L3/4椎间盘作为实验干预椎间盘进行手术处理,L1/2和L5/6椎间盘为空白对照椎间盘不进行处理。于术前随机选取2只动物进行腰椎核磁共振扫描;术后2、4、8、12周于每组随机选取2只动物行腰椎核磁共振扫描后处死并采集椎间盘,进行二甲基亚甲基蓝(DMMB)染色分光光度法髓核蛋白多糖含量测定,采用Pfirrmann分级方法对腰椎T2WI核磁共振图像分别进行退变程度的评估。实验结果显示: 1.经皮纤维环穿刺组动物无麻醉意外、伤口感染、后肢瘫痪等手术并发症发生,顺利存活至实验结束;右后外侧腹膜外入路纤维环穿刺组一只动物术中出现麻醉意外,导致死亡;右后外侧腹膜外入路髓核抽吸组一只动物术后出现切口感染、裂开,经换药、再次缝合和肌注抗生素后切口愈合。 2.空白对照椎间盘术后各时间点其椎间盘髓核内蛋白多糖含量、腰椎矢状位T2加权相核磁影像Pfirrmann分级评分基本保持一致。手术干预组椎间盘的退变程度均呈现逐渐加重趋势,髓核抽吸组于术后2周即出现可观察到的退变征象,并且在术后同一时间点其退变程度较纤维环穿刺方法构建的动物模型严重,纤维环穿刺方法构建的动物模型于术后4周才出现退变征象。术后同一检测时间点经皮纤维环穿刺法与右后外侧腹膜外入路纤维环穿刺法构建的椎间盘退变动物模型退变表现基本一致。髓核抽吸法可以快速建立椎间盘退变动物模型,而纤维环穿刺方法诱发的椎间盘退行性改变较缓慢。C形臂X射线机辅助下经皮纤维环穿刺方法可以替代传统的手术方法模拟椎间盘退变的影像学和生物化学改变,且手术创伤小、术后并发症少,是一种可行的椎间盘退变模型制备方法。
[Abstract]:The ideal animal model of intervertebral disc degeneration is a necessary condition for the research of intervertebral disc tissue engineering. However, there is still a lack of recognized model preparation methods. It has become an urgent task to explore a model preparation method of small animal wound and simple model operation.
The advantages and disadvantages of the C - arm assisted percutaneous fibrous ring puncture, the right posterolateral extraperitoneal approach and the right posterolateral extraperitoneal intramedullary aspiration for the preparation of the rabbit disc degeneration animal model were compared by biochemical and imaging methods. The C - arm assisted percutaneous fibrous ring puncture method was used to prepare the disc degeneration. The feasibility of changing animal models.
The experiment is divided into two parts. The first part is the establishment of the animal model of intervertebral disc degeneration, and the second part is the evaluation of the animal model of intervertebral disc degeneration.
27 New Zealand white rabbits were selected in the experiment. The experimental animals were randomly divided into the C - arm assisted percutaneous fibrous ring puncture group, the right posterolateral extraperitoneal approach group and the right posterolateral extraperitoneal intramedullary aspiration group with 9 animals in each group. Each New Zealand white rabbit was selected as the L2/3 and L3/4 intervertebral disc as an experimental intervertebral disc. The L1/2 and L5/6 intervertebral discs were blank and the intervertebral discs were not treated.
2 animals were randomly selected for lumbar NMR scan before operation. After 2,4,8,12 weeks, 2 animals in each group were randomly selected to die and collect intervertebral disc after lumbar spinal magnetic resonance scanning. Two methylmethylene blue (DMMB) colorimetric method was used to determine the concentration of nucleoprotein polysaccharide, and the T2WI nuclear magnetic resonance (NMR) of the lumbar vertebra was performed by Pfirrmann classification. The vibration images are evaluated for the degree of degeneration respectively.
The experimental results show that:
1. the animals had no anaesthesia, wound infection, posterior limb paralysis and other surgical complications and survived to the end of the experiment. The incision healed after dressing, suture and intramuscular injection of antibiotics.
2. the content of proteoglycan in the nucleus pulposus of the intervertebral disc at each time point after the intervertebral disc operation and the Pfirrmann grading score of the T2 weighted phase of the lumbar vertebral sagittal position were basically consistent. The degeneration degree of the intervertebral disc in the operation intervention group was gradually aggravated, and the degeneration signs of the nucleus pulposus group were observed at the 2 week after the operation, and In the same time point, the degree of degeneration was more serious than that of an animal model constructed by a fibrous ring puncture method. The animal model constructed by the method of fibrous ring puncture only appeared 4 weeks after the operation. The performance of the model is basically the same.
The method of nucleus pulposus aspiration can quickly establish the animal model of intervertebral disc degeneration, and the degeneration of the intervertebral disc induced by the fibrous ring puncture method is better than the slow.C arm X ray machine assisted percutaneous fibrous ring puncture method, which can replace the traditional surgical method to simulate the imaging and chemical changes of intervertebral disc degeneration, and the surgical trauma is small. It is a feasible method for the preparation of intervertebral disc degeneration models.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332

【参考文献】