Kong Jie, Wang Zi-xuan, Ji Ai-yu, Wang De-chun, Qi Zong-hua, Xu Wen-jian, Hao Da-peng, Duan Feng, Hu You-gu
Department of Spinal Surgery, the Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, China.
Zhonghua Wai Ke Za Zhi. 2008 Jun 1;46(11):835-8.
To establish a novel model of lumbar disc degeneration on the early stage in the rhesus monkey using percutaneous needle puncture guided by CT.
(1) Thirteen rhesus monkeys aged from 4 to 7 years, female 7 and male 6 were selected for establishing a model of the early stage of lumbar disc degeneration. (2)13 monkeys, 91 discs were divided into 3 groups: 64 discs from L1/2 to L5/6 were percutaneous punctured with a needle 20G as experimental group and 1 disc with a needle 15G as puncture control group and 26 discs were not be punctured from L6,7 to L7-S1 as control group. (3) Lumbar disc localization for needle puncture was guided by CT. All discs were examined by MRI, the HE, Masson's trichrome, Safranine-O and immunohistochemical staining of type II collagen before disc puncture and after puncture at 4, 8 and 12 weeks.
MRI: (1) Experimental group: Pfirmann's Grade I was shown at postoperation 4, 8 and 12 weeks; (2) Puncture control group: Grade III was shown at postoperation 4 weeks and Grade IV at 8 weeks; (3) CONTROL GROUP: Grade I was shown at postoperation 4, 8 and 12 weeks. Histological examination: (1) In experimental group, there was no any change at postoperation 4 weeks, and the cell population of the nucleus was decreased at 8 weeks and more decreased at 12 weeks in HE. (2) There was no any change at postoperation 4 weeks, the clefts among the lamellae of the annulus fibrosus (AF) were shown at 8 weeks and more wider of the clefts of AF at 12 weeks in Masson's trichrome. (3) No any change was shown at postoperation 4 weeks, proteoglycan were progressively decreased at 8 and 12 weeks in Safranine-O. (4) No statistically significant difference in positive rate was observed at 4 and 8 weeks compared with control group in immunohistochemical staining of type II collagen. There was statistical difference at 12 weeks compared with control group (P<0.05). In puncture control group postoperation 8 weeks, the morphology of cell of nucleus pulposus was not clear in HE. The wider clefts of lamellae of the AF were shown in Masson's trichrome. The proteoglycan was obviously decreased in Safranine-O. Immunohistochemical staining collagen II synthesized was decreased. In normal control group, no any change was shown at 4, 8 and 12 weeks.
The degeneration of lumbar intervertebral disc on the early stage could be induced by the percutaneous needle puncture (20G) to the annulus fibrosus. The assessment of disc degeneration on early stage is not shown on MRI and only confirmed by histological examination.
利用CT引导下经皮穿刺法，在恒河猴身上建立一种新型的早期腰椎间盘退变模型。
（1）选取13只4至7岁的恒河猴，其中雌性7只，雄性6只，用于建立早期腰椎间盘退变模型。（2）将13只猴子的91个椎间盘分为3组：L1/2至L5/6的64个椎间盘用20G针经皮穿刺作为实验组，1个椎间盘用15G针穿刺作为穿刺对照组，L6、7至L7-S1的26个椎间盘不穿刺作为对照组。（3）经皮穿刺的腰椎间盘定位由CT引导。所有椎间盘在穿刺前及穿刺后4周、8周和12周均进行MRI、HE、Masson三色染色、番红O染色及II型胶原免疫组化染色检查。
MRI检查结果：（1）实验组：术后4周、8周和12周均显示为Pfirmann I级；（2）穿刺对照组：术后4周显示为III级，8周显示为IV级；（3）对照组：术后4周、8周和12周均显示为I级。组织学检查结果：（1）实验组，HE染色显示术后4周无任何变化，8周时髓核细胞数量减少，12周时进一步减少。（2）Masson三色染色显示术后4周无任何变化，8周时纤维环板层间出现裂隙，12周时纤维环裂隙更宽。（3）番红O染色显示术后4周无任何变化，8周和12周时蛋白聚糖逐渐减少。（4）II型胶原免疫组化染色在4周和8周时与对照组相比阳性率无统计学差异。12周时与对照组相比有统计学差异（P<0.05）。穿刺对照组术后8周，HE染色显示髓核细胞形态不清。Masson三色染色显示纤维环板层裂隙更宽。番红O染色显示蛋白聚糖明显减少。免疫组化染色显示II型胶原合成减少。正常对照组在4周、8周和12周均无任何变化。
经皮穿刺（20G）纤维环可诱导早期腰椎间盘退变。早期椎间盘退变在MRI上未显示，仅通过组织学检查得以证实。
