中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (5): 995-1002.doi: 10.12307/2025.249

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

X射线透视引导下不同方式建立兔椎间盘退变模型的结果对比

丁至立1，2，3，黄  杰3，蒋  强3，李土胜3，刘  江1，2，3，丁  宇1，2，3   

  1. 1安徽医科大学海军临床学院，北京市  100048；2安徽医科大学第五临床医学院，安徽省合肥市  230032；3解放军总医院第六医学中心中医医学部骨伤科，北京市  100048
  • 收稿日期:2023-12-22 接受日期:2024-02-22 出版日期:2025-02-18 发布日期:2024-06-03
  • 通讯作者: 丁宇，教授，主任医师，博士研究生导师，安徽医科大学海军临床学院，北京市 100048；安徽医科大学第五临床医学院，安徽省合肥市 230032；解放军总医院第六医学中心中医医学部骨伤科，北京市 100048
  • 作者简介:丁至立，男，1998年生，汉族，硕士，主要从事微创脊柱外科、分子生物学方面的研究。
  • 基金资助:
    国家自然科学基金资助项目(82274637)，项目负责人：丁宇

Constructing rabbit intervertebral disc degeneration models by different methods under X-ray guidance: a comparative study 

Ding Zhili1, 2, 3, Huang Jie3, Jiang Qiang3, Li Tusheng3, Liu Jiang1, 2, 3, Ding Yu1, 2, 3   

  1. 1Naval Clinical College, Anhui Medical University, Beijing 100048, China; 2The Fifth Clinical College of Anhui Medical University, Hefei 230032, Anhui Province, China; 3Department of Orthopedics and Traumatology, Division of Traditional Chinese Medicine, Sixth Medical Center, PLA General Hospital, Beijing 100048, China
  • Received:2023-12-22 Accepted:2024-02-22 Online:2025-02-18 Published:2024-06-03
  • Contact: Ding Yu, Professor, Chief physician, Doctoral supervisor, Naval Clinical College, Anhui Medical University, Beijing 100048, China; The Fifth Clinical College of Anhui Medical University, Hefei 230032, Anhui Province, China; Department of Orthopedics and Traumatology, Division of Traditional Chinese Medicine, Sixth Medical Center, PLA General Hospital, Beijing 100048, China
  • About author:Ding Zhili, Master, Naval Clinical College, Anhui Medical University, Beijing 100048, China; The Fifth Clinical College of Anhui Medical University, Hefei 230032, Anhui Province, China; Department of Orthopedics and Traumatology, Division of Traditional Chinese Medicine, Sixth Medical Center, PLA General Hospital, Beijing 100048, China
  • Supported by:
    the National Natural Science Foundation of China, No. 82274637 (to DY)

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摘要：




文题释义：
纤维环穿刺法：一种经典的椎间盘退变模型建立方法，使用18G注射针头于椎板间隙平行终板方向刺入椎间盘中央，维持5 s后拔出穿刺针。
无水乙醇终板注射法：一种适用于小型动物的椎间盘退变建立方法，使用16G注射针头在L2/L3、L3/L4、L4/L5上下终板注射50 μL无水乙醇。

背景：国内外学者认为新西兰大白兔腰椎解剖形态与人类腰椎更为相似，是理想的模型动物。在X射线引导下采用不同方式建立兔椎间盘退变模型目前尚缺乏系统性比较。
目的：基于X射线引导采用针刺法、终板注射法以及改良终板注射法建立兔腰椎间盘退变模型，比较3种方法造模的效果。
方法：6月龄新西兰大白兔16只随机分为4组，分别为针刺组、终板注射组、改良终板注射组及空白对照组，针刺组对3个连续节段(L2/3、L3/4、L4/5)椎间盘进行针刺造模，终板注射组在3个连续节段终板单点注射50 μL无水乙醇，改良终板注射组对3个连续节段椎间盘进行针刺并在相应节段终板4个方位点注射50 μL无水乙醇，空白对照组不进行干预。术后 2，4，8 周进行X射线检查，测量椎间隙高度指数，然后取椎间盘组织进行解剖学观察、病理学检查。
结果与结论：①解剖学观察显示，针刺组以纤维环破溃为主要表现，终板注射组主要以髓核退变为主要表现，改良终板注射组以全椎间盘结构紊乱为主要表现；②与空白对照组相比，针刺组术后2周椎间隙高度指数降低最明显，终板注射组术后2，4周椎间隙高度指数降低明显，改良终板注射组术后2，4，8周椎间隙高度指数均显著降低；③病理学检查显示，针刺组纤维环结构出现破损，内层纤维环向内凸入，终板注射组表现为终板裂隙、排列混乱及细胞核丢失，改良终板注射组表现为整体椎间盘结构紊乱，髓核失水皱缩，与破损的纤维环无明显边界。结果表明，针刺法、终板注射法及改良终板注射法均能建立兔椎间盘退变模型，改良终板注射法较单一方法可大幅加快并加重椎间盘退变，可有效缩短实验周期。
https://orcid.org/0009-0001-0576-7978(丁至立)

关键词: 组织工程, 椎间盘退变, 纤维环穿刺法, 终板注射法, X射线, 新西兰大白兔

Abstract: BACKGROUND: Scholars at home and abroad consider New Zealand rabbits to be an ideal model animal because of the similar anatomical morphology of the lumbar spine to that of the human lumbar spine. There is a lack of systematic comparison of different ways to establish rabbit intervertebral disc degeneration models under X-ray guidance.
OBJECTIVE: To establish a rabbit model of lumbar disc degeneration using X-ray guided acupuncture, end-plate injection and combined method, and to compare the modeling effects of these three methods.
METHODS: Eighteen 6-month-old New Zealand white rabbits were randomly selected and divided into four groups: acupuncture group, endplate injection group, combined group and blank control group. In the acupuncture group, three consecutive segments of the intervertebral discs (L2/3, L3/4, L4/5) were needled and modeled; in the endplate injection group, 50 μL of anhydrous ethanol was injected at a single point on the endplates of the three consecutive segmental discs; in the combined group, three consecutive segmental intervertebral discs were needled and injected with 50 μL of anhydrous ethanol at four azimuthal points on the endplates of the corresponding segmental discs; and the blank control group received no interventions. X-ray examination was performed to measure the disc height index at 2, 4, and 8 weeks after surgery. The intervertebral disc tissues were then taken for anatomical observation and pathological examination. 
RESULTS AND CONCLUSION: (1) Anatomical examination showed that fibrous annulus rupture, nucleus pulposus degeneration, and total disc structure disorder were the main manifestation in the acupuncture group, endplate injection group, and combined group, respectively. (2) X-ray examination showed that the disc height index showed the most obvious reduction in the acupuncture group at 2 weeks after operation, significant reductions in the endplate injection group at 2 and 4 weeks after operation, and significant reductions in the combined group at 2, 4, and 8 weeks after operation. (3) Pathological examination showed that the fibrous ring structure was damaged and the inner annulus fibrosus protruded inward in the acupuncture group; endplate fissure, disordered arrangement and nucleus loss were observed in the endplate injection group; total disc structure disorder with the nucleus pulposus losing water and shrinking and no obvious border with the broken annulus fibrosus was found in the combined group. To conclude, acupuncture, endplate injection and the modified endplate injection method can establish the rabbit intervertebral disc degeneration model. Compared with the single method, the modified endplate injection method can greatly accelerate and aggravate the degeneration of the intervertebral disc, and can effectively shorten the experiment period.


Key words: tissue engineering, intervertebral disc degeneration, annulus fibrosus puncture, endplate injection, X-ray, New Zealand white rabbit

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