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目的:比较体外培养的退变髓核细胞与正常髓核细胞生物学特性的差异。方法:将成年新西兰大白兔以纤维环穿刺法椎间盘造模法处理,4周后取退变以及正常髓核细胞,进行细胞形态及电镜观察、MT法试验,检测细胞周期及主要细胞外基质相关基因表达情况。结果:细胞生长曲线显示,传4代细胞第5天开始出现生长减慢,传5～7代细胞均在接种后第2～3天细胞数量明显减少(P<0.05);退变髓核细胞传3代细胞第4天开始出现生长减慢(P<0.05),传5代时即出现生长滞缓,传7代时出现生长停滞。髓核软骨样细胞MTT摄取实验,传4～7代细胞的吸光度值明显低于传1代(P<0.05)。正常和退变髓核细胞细胞所处生长分裂期对比中,正常和退变髓核细胞G1期细胞所占比例具有统计学差异(P<0.05)。结论:大白兔正常髓核细胞持续增殖能力、蛋白聚糖和Ⅱ型胶原DNA的含量均较退变髓核细胞高,能为椎间盘组织工程学、细胞生物学疗法等提供理论基础。
OBJECTIVE: To compare the biological characteristics of degenerative and normal nucleus pulposus cells cultured in vitro. Methods: The adult New Zealand white rabbits were treated by fibrocapsule intervertebral disc modeling. Degeneration and normal nucleus pulposus cells were obtained after 4 weeks. Morphology and electron microscopy were observed. MT assay was used to detect cell cycle and major extracellular matrix Gene expression. Results: The cell growth curve showed that the passage 4 cells began to slow down on the 5th day, and the number of passage 5-7 passage cells decreased significantly (P <0.05) on the 2nd to 3rd day after inoculation; degenerative nucleus pulposus cells On the 4th day, the passage 3 cells began to slow down (P <0.05), and the growth retardation appeared on the 5th passage, and the growth arrest occurred on the 7th passage. Nucleus pulposus cartilage-like cells MTT uptake test, pass 4 to 7 generations of cells absorbance value was significantly lower than the pass generation (P <0.05). In normal and degenerative nucleus pulposus cells, the proportion of G1 phase cells in normal and degenerated nucleus pulposus cells was significantly different (P <0.05). CONCLUSION: The normal proliferative capacity of normal nucleus pulposus cells in rabbits is higher than that in degenerated nucleus pulposus cells, which provides a theoretical basis for tissue engineering and cell biology therapy.