体外培养人退变髓核与纤维环细胞增殖能力的研究

目的 探讨体外培养的人退变髓核与纤维环细胞的增殖能力,比较2种退变细胞体外培养中所表现的不同生物学行为,为椎间盘退变疾病的预防与治疗提供新的理论依据.方法 采集临床腰椎间盘突出症患者手术取材的椎间盘组织标本,病理学诊断评估其退变程度,酶消化法原代培养髓核与纤维环细胞,并鉴定.每例病例的髓核细胞与纤维环细胞分别体外培养至第5代,各代次细胞传代接种密度控制为1×105个.同条件培养48 h后,观察每一代细胞的形态学变化,同时应用流式细胞仪检测2种细胞的增殖能力.结果 体外培养的退变髓核与纤维环细胞生长状态良好.随着体外传代的进行,退变髓核与纤维环细胞S期细胞百分比和增殖指数(PI)均呈上升趋势,髓核细胞PI值于第3代时达到峰值,而纤维环细胞PI值于第5代时最高;且第2～4代的退变髓核细胞的增殖活性均高于退变纤维环的增殖活性(P＜0.05).结论 体外培养的人退变髓核与纤维环细胞有着不同的增殖特点,髓核与纤维环细胞对体内退变微环境有着不同的响应机制,并影响着整个椎间盘退变的进展.

Study of the proliferation ability of human degenerate nucleus pulposus and annular fibrosus cell in vitro

Objective To explore the proliferation of human degenerate nucleus pulposus and annular fibrosus cell in vitro and compare the different biological behavior between the two kinds of cell after degeneration, and provide the new theoretical basis for the prevention and treatment of degenerative disc disease. Methods The samples of intervertebral disc tissue were collected from patients with lumbar disc herniation. The degree of degeneration was assessed by the pathological diagnosis and degenerate nucleus pulposus and annular fibrosus cell were cultured by enzymatic digestion and identified. In each case, the control groups of the nucleus pulposus cells and annulus fibrosus cells were cultured to the fifth generation. The inoculation density of cells was 1 xl05. The cell morphology of each generation was observed, while the proliferation of cells was detected by flow cytometry after 48h culture with the same conditions. Results The degenerate nucleus pulposus cell and annular fibrosus cell were in good condition in vitro. The percentages of S phase cell and proliferation index （PI） were both on the rise with the subculture. The PI of nucleus pulposus cells reached the peak in the 3rd generation; The PI of annulus cells was the highest in the generation 5. The proliferation activity of degenerate nucleus pulposus cell in 2-4 generations was higher than the degenerate annular fibrosus cell within the same generations （P〈0.05）. Conclusion Different proliferative characteristics of the degenerate nucleus pulposus and annular fibrosns cell confirmed that the disc degeneration is reversible. The response mechanisms to the degenerate micro-environment are completely different in vivo and affects the entire disc degeneration progress.