兔髓核与纤维环细胞生物学特性差异的研究  

Different biological characteristics between nucleus pulposus and annulus fibrosus cells in rabbits

作　　者：谢健[1] 童培建[2] 肖鲁伟[1] 金红婷[1] 吴承亮[1] 单乐天[1] 毛强[1] 潘佳菲 

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江省中医院骨科,浙江杭州310006

出　　处：《中国骨伤》2013年第6期481-485,共5页China Journal of Orthopaedics and Traumatology

基　　金：2009年浙江省自然科学基金(编号:Y2091189);浙江省医药卫生科技计划项目(编号:2010KYA142)~~

摘　　要：目的:同时建立兔髓核细胞与纤维环细胞体外培养模型,比较两者生物学特性差异。方法:新西兰大白兔5只(2～3kg,雌雄不限),无菌条件下分离髓核及纤维环,酶消化法联合组织块法含15%FBS的DMEM/F12(1∶1)培养液培养,当细胞90%融合后进行传代培养。通过倒置相差显微镜观测细胞形态,台盼蓝染色测定细胞活力,甲苯胺蓝和HE染色进行组织学观察,MTT法测定细胞增殖,分析比较髓核细胞与纤维环细胞形态、活力、增殖的差异。结果:原代及第1代髓核细胞和纤维环细胞形态上无明显差异,第2代髓核细胞开始有空泡出现。髓核细胞较纤维环细胞贴壁时间晚、细胞活力低;原代髓核细胞从第9天开始增殖速度较纤维环细胞慢(P<0.05)。结论:纤维环细胞的细胞活性明显高于髓核细胞。椎间盘退变性疾病可能是由髓核发生衰退引起,从而局部生物力学改变,导致纤维环破裂等组织结构的改变及功能的丢失。Objective：To compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model. Methods：Five New Zealand white rabbits（2 to 3 kg,either gender） were isolated nucleus pulposus and annulus fibrosus under sterilized condition,then cultured in nutrient solution with 15% FBS and DMEM / F12 （1∶1） by enzyme digestion combined with tissue block method. When 90% cells fused,subcultring were performed. Cell morphology were observed by inverted phase contrast microscope,cell viability were detected by trypan blue staining,histological were observed by a toluidine blue and HE staining,cell proliferation were tested by MTT method,then the cell morphology,viability,proliferation between nucleus pulposus and annulus fibrosus were compared. Results：There were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later,and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day. Conclusion：The cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus ,local biomechnical changes,furether caused structure change and function loss of annulus fibrosus.

关 键 词：兔 椎间盘 细胞培养 生物差异 

分 类 号：R589.2[医药卫生—内分泌]