BACKGROUND: Hydrostatic pressure stimulation caused by gravity is an important regulator of human nucleus pulposus cells metabolism.OBJECTIVE: To observe the effect of hydrostatic pressure on the morphology of monolayer cultured human nucleus pulposus cells and matrix expression in vitro.METHODS: In the hydrostatic pressure loading system, 0.3, 0.7, 3 Mpa pressure were imposed on the passage 4 human nucleus pulposus cells in monolayer in vitro culture for 30, 60, 90, 120 minutes, and the atmospheric pressure 0.1 Mpa group was as the control group.RESULTS AND CONCLUSION: ①Morphology: cell volume became small under hydrostatic pressure stimulation. Under 0.3and 0.7 Mpa hydrostatic pressure, cell volume shrinked slightly, and cell morphology were relatively complete. Under 3 Mpa hydrostatic pressure, cell volume was significantly reduced, and cell morphology was not complete. ②Survival rate: after 30 minutes hydrostatic pressure stimulation, survival rate were relatively low in all groups, as time passed, the rate gradually increased or remained stably in 0.3, 0.7 Mpa groups, and tended to decline in 3 Mpa group.③ Proteoglycan expression: the expression were increasing with time in all groups, and when hydrostatic pressure sustained for 120 minutes, the strongest proteoglycan expression were detected in 0.3 and 0.7 Mpa group which were significantly more than that in the 0.1 and 3 Mpa group. The results show that hydrostatic pressure would affect nucleus pulposus cell morphology, viability and proteoglycan expression.%背景:加载在椎间盘上的重力引起的静态压力刺激是椎间盘细胞代谢的重要调节因素.目的:观察静水压对体外单层培养人椎间盘髓核细胞形态学及基质代谢的影响.方法:在静水压加载系统中对体外单层培养的传4 代人髓核细胞施以0.3,0.7,3 MPa 的静压,分别加压30,60,90,120 min,以常压0.1 MPa 为对照.结果与结论:①髓核细胞形态:在静水压干预下细胞体积均变小.0.3,0.7 MPa 静水压下轻微缩小,细胞形态相对完整;3 MPa 静水压下缩小最明显,且细胞形态不完整.②髓核细胞存活率:在持续静水压刺激下开始的30 min,不论压力大小存活率都偏低,在0.3,0.7 MPa 时随作用时间延长而增加或维持稳定,细胞增殖逐渐增强,在3 MPa 时随时间趋于下降,最终细胞总体数量减少.③髓核细胞蛋白多糖:各组表达量随时间增加逐渐增加,当持续加压120 min 时,在0.3,0.7 MPa静水压下合成量呈高表达状态,在0.1,3 MPa 静压表达相对较少.表明静水压会对髓核细胞形态学、存活率及基质表达产生影响.
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