Establishing an intervertebral disc degeneration model using annulus puncture

doi:10.3969/j.issn.2095-4344.2017.12.009

Abstract

Abstract:

BACKGROUND: Annulus puncture method is commonly reported in the establishment of intervertebral disc degeneration (IDD) models, but there is a lack of comparative studies concerning complete MRI, pathologyical and extracellular matrix changes. Thereafter, a further exploration is imperative.
OBJECTIVE: To investigate the feasibility of establishing IDD models using annulus puncture.
METHODS: Thirty New Zealand white rabbits were randomly divided into two groups (n=15 per group). In experimental group, through extraperitoneal approach, the L1-2, L2-3, and L3-4 intervertebral space were exposed, and then stabbed by 16-gauge hypodermic needles; while rabbits in sham operated group were only subjected to making and incision but not to puncture. Five rabbits in each group were randomly selected for MRI examination at 4, 8 and 16 weeks after modeling, and were sacrificed through air embolism to obtain the nucleus pulposus. Histological changes were observed by hematoxylin-eosin staining, and the level of proteoglycan was detected by Safranin O staining.
RESULTS AND CONCLUSION: In the experimental group, T2-weighted signal in the disc was decreased in a time-dependent manner; an obvious reduction was found at 4 weeks after modeling, and reached the peak at 16 weeks. In contrast, the sham operated group showed no significant changes at each time point. Hematoxylin-eosin staining showed that in the experimental group, cartilage-like cells decreased, while fibroblast-like cells in the disc increased in number and proportion, and extracellular fibers were in disorder; all above changes became obvious at 4 weeks after surgery and the most obvious at 16 weeks, but there were no significant changes in the sham operated group. Safranin O staining revealed that the color of the tissues under puncture became lighter, and this variation became overt at 4 weeks after surgery and the most obvious at 16 weeks. In conclusion, annulus puncture is a reliable method to make IDD models.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Intervertebral Disk Degeneration, Models, Animal, Tissue Engineering

CLC Number:

R318

Wang Zi-xing, Wei Peng. Establishing an intervertebral disc degeneration model using annulus puncture[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(12): 1855-1860.

Figures/Tables 4

References

[1] Freemont AJ. The cellular pathobiology of the degenerate intervertebral disc and discogenic back pain. Rheumatology (Oxford). 2009;48:5-10.

[2] Masuda K. Biological repair of the degenerated intervertebral disc by the injection of growth factors. Eur Spine J. 2008;17 Suppl 4: 441-451.

[3] Masuda K, An HS. Growth factors and the intervertebral disc.Spine J. 2004;4(6 Suppl):330S-340S.

[4] 谭伟伟,何升华,孙志涛,等.微创针刺旋切制备兔椎间盘退变模型[J].中国修复重建外科杂志,2016,3(30):343-347.

[5] 崔运能,李绍林,周荣平, 等.CT引导下经皮纤维环穿刺建立兔腰椎间盘退变模型[J]. 中国脊柱脊髓杂志,2016,24(3): 234-243.

[6] Issy AC, Castania V, Castania M, et al. Experimental model of intervertebral disc degeneration by needle puncture in Wistar rats. Braz J Med Biol Res. 2013;46(3):235-244.

[7] 王珏,张雷,刘杨青,等．经皮微创纤维环穿刺法制作兔椎间盘退变模型[J]. 第三军医大学学报,2013,35(15):1579-1582.

[8] 邵辉,卞正君,孙建华.纤维环穿刺髓核抽吸法建立兔椎间盘退变模型[J].石河子大学学报,2015,33(1):97-101.

[9] 白亦光,韩小伟,张旭乾,等.纤维环穿刺抽吸法和纤维环切开法建立兔椎间盘退变模型的比较[J].中国矫形外科杂志,2013, 21(7):679-694.

[10] 罗平,刘玉林,陈仲,等. 纤维环穿刺法与纤维环切开法建立兔椎间盘退变模型[J].中国组织工程研究与临床康复,2010, 11(14):1955-1958.

[11] 杨效宁,孙天威,王启明,等. 纤维环穿刺法诱导兔椎间盘退变模型的组织学和影像学分析[J]. 徐州医学院学报,2010, 30(5):314-317.

[12] 吴剑宏,阮狄克.腰椎间盘退变的MRI诊断分级及其临床应用进展[J].中国脊柱脊髓杂志,2010,20(6):511-513.

[13] Silberberg R. The vertebral column of diabetic sand rats (psammomys obesus). Exp Cell Biol. 1988;56(4): 217-220.

[14] 吴靖平,陈统一,陈中伟,等.双后肢大鼠椎间盘退变动物模型的建立[J].中华实验外科杂志, 2004, 21(1): 105-107.

[15] Clouel J, Pot-Vaucel M, Grimandi G, et al. Characterization of the age-dependent intervertebral disc changes in rabbit by correlation between MRI, histology and gene expression. BMC Musculoskelet Disorrl. 2011;12: 147.

[16] Lotz JC. Animal models of intervertebral disc degeneration: Lessons learned. Spine. 2004;29(23): 2742-2750.

[17] Kroeber MW, Unglaub F, Wang H, et al. New in vivo animal model to create intervertebral disc degeneration and toinvestigate the effects of therapeutic strategies to stimulate disc regeneration. Spine. 2002;27(23): 2684-2690.

[18] 熊蠡茗,邵增务,郭兵,等.可控轴向压力致兔腰椎间盘退变模型的建立及评价[J].中国矫形外科杂志, 2009, 17(19):1492- 1496.

[19] 黄宗强,刘尚礼,郑召民.双侧关节突关节切除诱发椎间盘退变的磁共振计量分析[J].中国矫形外科杂志, 2007, 15(15): 1175-1177.

[20] Imai Y, Okuma M, An HS et al. Restoration of disc height loss by recombinant human osteogenic protein-1 injection into intervertebral discs undergoing degeneration induced by an intradiscal injection of chondroitinase ABC. Spine. 2007;32(11): 1197-1205.

[21] Radcliff KE, Kepler CK, Jakoi A,et al. Adjacent segment disease in the lumbar spine following different treatment interventions. Spine J. 2013;13(10): 1339-1349.

[22] Kepler CK, Ponnappan RK, Tannoury CA,et al. The molecular basis of intervertebral disc degeneration．Spine J.2013;13(3): 318-330.

[23] Cho H,Lee S,Park SH,et al. Synergistic effect of combined growth factors in porcine intervertebral disc degeneration. Connect Tissue Res. 2013;54(3):181-186.

[24] Lee H, Sowa G, Vo N, et al. Effect of bupivacaine on intervertebral disc cell viability.Spine J. 2010;10(2)：159-166.

[25] Haschtmann D,Stoyanov JV,Gedet P,et al. Vertebral endplate trauma induces disc cell apoptosis and promotes organ degeneration in vitro. Eur Spine J. 2008;17(2): 289-299.

[26] Sun W, Zhang K, Liu G, et al. Sox9 gene transfer enhanced regenerative effect of bone marrow mesenchymal stem cells on the degenerated intervertebral disc in a rabbit model. PLoS One. 2014;9(4):e93570.

[27] Xia X, Guo J, Lu F, et al. SIRT1 plays a protective role in intervertebral disc degeneration in a puncture induced rodent model. Spine(Phila Pa1976). 2015;40(9): E515-E524.

[28] 江立波,张小磊,徐华梓,等.细胞自噬对饥饿环境下椎间盘髓核细胞的保护作用[J].中国病理生理杂志,2012,28(7):1302- 1307.

[29] 郑旭浩,张小磊,江立波,等．细胞自噬在糖尿病大鼠椎间盘退变中的作用[J]. 中国病理生理杂志,2013,29(11):2011-2016.

[30] 杜炎鑫,佟德民,邓晋丰,等.纤维环穿刺法建立兔腰椎间盘退变模型[J]. 宁夏医科大学学报,2013,5(20):132-135.

[31] Sobajima S, Kompel JF, Kim JS, et al. A slowly progressive and reproducible animal model of intervertebral disc degeneration characterized by MRI, X-ray, and histology. Spine (Phila Pa 1976). 2005;30(1): 15-24.

[32] 黑龙,张佳林,李宏辉,等.经皮纤维环穿刺和纤维环切开法建立兔椎间盘退变模型的比较[J]．宁夏医科大学学报,2015,37(11): 1279-1282.

[33] Sobajima S, Kompel JF, Kim JS, et al. A slowly progressive andreproducible animal model of intervertebral disc degeneration characterized by MRI, X- ray, and histology. Spine. 2005;30(1): 15-24.

[34] 王诗军,李淳德,孙浩林,等.针刺损伤兔椎间盘退变模型组织病理变化及软骨样细胞的迁移规律[J].中国组织工程研究与临床康复,2012,9 (16):1597-1600.

[35] Longo UGL, Ripalda P, Denaro V, et al. Morphologic comparison of cervical, thoracic, lumbar intervertebral discs of cynomolgus monkey (Macaca fascicularis). Eur Spine J.2006;15(12): 1845-1851.