| 兔椎间盘器官与脊柱运动节段离体培养条件下髓核组织的变化 |
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Received:October 20, 2014
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| 作者 | Author | 单位 | Unit | E-Mail |
| 朱立国 |
ZHU Li-guo |
中国中医科学院望京医院脊柱二科, 北京 100102 |
The Second Department of Spine, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China |
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| 展嘉文 |
ZHAN Jia-wen |
中国中医科学院望京医院脊柱二科, 北京 100102 |
The Second Department of Spine, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China |
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| 冯敏山 |
FENG Min-shan |
中国中医科学院望京医院脊柱二科, 北京 100102 |
The Second Department of Spine, Wangjing Hospital of China Academy of Traditional Chinese Medicine Sciences, Beijing 100102, China |
fengminshan@hotmail.com |
| 张平 |
ZHANG Ping |
中国中医科学院望京医院病理科, 北京 100102 |
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| 朱岩 |
ZHU Yan |
中国中医科学院望京医院病理科, 北京 100102 |
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| 朱宏伟 |
ZHU Hong-wei |
中国中医科学院望京医院病理科, 北京 100102 |
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| 期刊信息：《中国骨伤》2015年28卷，第9期，第824-831页 |
| DOI：10.3969/j.issn.1003-0034.2015.09.011 |
| 基金项目:北京市科技专项中医正骨技术项目(编号:Z131107002813046) |
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目的:比较离体培养的兔椎间盘器官及脊柱运动节段两种模型椎间盘髓核组织的变化.
方法:将21只新西兰白兔随机分为器官组8只,节段组13只,处死后在无菌条件下分别取出椎间盘器官和脊柱运动节段各50个,在高渗培养基中进行整体培养(410 mOsm/kg),于培养前及培养后第3、7、14、21天,两组各取10个椎间盘分别进行HE染色、II型胶原免疫组化、蛋白多糖含量和髓核细胞活力测定.
结果:培养21 d器官组与培养14 d节段组HE染色示椎间盘组织结构基本保持完整,21 d节段组椎间盘组织形态学破坏;21 d器官组与14 d节段组Ⅱ型胶原免疫组化染色强度差异无统计学意义(P>0.05),21 d节段组染色变浅,与之前各时间点及器官组相比差异有统计学意义(P<0.05);蛋白多糖PAS/AB染色7 d内两组强度无降低,14 d两组强度均有所减弱,21 d节段组染色强度进一步减弱,改变比器官组更为明显;髓核细胞荧光检测两组7 d时强度较培养前变化不明显(P>0.05),21 d器官组与14 d节段组强度略有降低,但与之前时间点比较差异无统计学意义(P>0.05),21 d节段组髓核细胞荧光强度减弱,细胞活性降低,与之前各时间点及器官组比较差异明显(P<0.05).
结论:14d内脊柱运动节段可作为研究生物力学对椎间盘影响的离体实验模型. |
| [关键词]:椎间盘 脊柱 器官培养技术 兔 |
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| Changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segmen |
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Abstract:
Objective: To compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.
Methods: Twenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen typeⅡ, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.
Results: HE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group, but there was severely degenerated of 21 days segment group. The intensity value of type Ⅱcollagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group(P>0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days(P<0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups(P>0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P>0.05), however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased, and there was a significant difference compared with before each time points and organ group at 21 days(P<0.05).
Conclusion: It is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebraldisc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days. |
| KEYWORDS:Intervertebral disc Spine Organ culture techniques Rabbits |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 朱立国,展嘉文,冯敏山,张平,朱岩,朱宏伟.兔椎间盘器官与脊柱运动节段离体培养条件下髓核组织的变化[J].中国骨伤,2015,28(9):824~831 |
| 英文格式: | ZHU Li-guo,ZHAN Jia-wen,FENG Min-shan,ZHANG Ping,ZHU Yan,ZHU Hong-wei.Changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segmen[J].zhongguo gu shang / China J Orthop Trauma ,2015,28(9):824~831 |
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