渗透压负荷对兔椎间盘器官培养模型的影响

发布时间：2018-11-13 07:31

【摘要】：目的建立一种可用于体外研究椎间盘退变的椎间盘器官培养模型,探讨渗透性负荷对模型椎间盘细胞活力和代谢的影响。方法4～6月龄新西兰大白兔10只,均分为两组,处死后立即手术切取胸腰段椎间盘,每只9个,分别在等渗(300 mOsm/kg,等渗组)或高渗(410 mOsm/kg,高渗组)培养基中进行整体器官培养,在培养前和培养后的第7、14、21、28天,利用Mitotracker Green荧光探针、组织化学和生物化学方法评估两组椎间盘髓核细胞的活力、结构的完整性以及蛋白多糖含量的变化。结果取材后培养前椎间盘髓核细胞的荧光强度最高,为11503±402,在体外培养过程中,高渗组第14天荧光强度为9202±907,与培养前比较差异有显著性意义(P0.05),第7、21和28天分别为10504±710、10860±711、10713±953;等渗组第7、14、21、28天分别为11350±351、11207±385、10914±300、10862±229,与培养前相比较无显著性差异(P0.05),两组在第7、21和28天时无显著性意义(P0.05)。在培养过程中,两组椎间盘髓核和纤维环的组织结构能够基本保持完整。髓核蛋白多糖含量在培养的第7天高渗组和等渗组分别为3.33±.028和2.83±0.25,均较培养前(5.03±0.37)明显降低(P0.01),第14、21、28天时与第7天相比下降不明显(P0.05),两组相同时间点的差异无显著性意义(P0.05)。结论椎间盘器官培养模型可以在等渗或高渗环境中有效维持兔椎间盘结构的完整性和髓核细胞的活力至少4周。
[Abstract]:Objective to establish an organ culture model of intervertebral disc degeneration in vitro and to explore the effect of osmotic loading on the cell viability and metabolism of intervertebral disc. Methods Ten New Zealand white rabbits aged 4 to 6 months were divided into two groups. The thoracolumbar intervertebral discs were removed immediately after death, each of 9 rabbits were divided into isosmotic group (300 mOsm/kg, isosmotic group) or hyperosmotic group (410 mOsm/kg,). The whole organ was cultured in the medium of hyperosmotic group. The viability of nucleus pulposus cells in the two groups was evaluated by Mitotracker Green fluorescence probe, histochemical and biochemical methods before and after culture. Structural integrity and changes in proteoglycan content. Results the fluorescence intensity of nucleus pulposus cells was the highest (11503 卤402) before culture. The fluorescence intensity in hyperosmotic group was 9202 卤907 on the 14th day, which was significantly different from that before culture (P0.05). On the 21st and 28th days, it was 10504 卤71010860 卤711713 卤953, respectively. In the isosmotic group, it was 11350 卤351U 11207 卤385U 10914 卤300m 10862 卤229on the 28th day, no significant difference was found between the two groups (P0.05), but there was no significant difference between the two groups on the 7th day and 28th day (P0.05). The tissue structure of the nucleus pulposus and the fibrous ring of the intervertebral disc were basically intact during the culture. The contents of myelin proteoglycan in hyperosmotic group and isosmotic group were 3.33 卤0.28 and 2.83 卤0.25 on the 7th day, respectively, which were significantly lower than those before culture (5.03 卤0.37) (P0.01). There was no significant difference between the two groups in the same time point (P0.05) at the 28th day after the 14th day (P0.05) and the 7th day (P0.05). Conclusion the model of intervertebral disc organ culture can effectively maintain the integrity of intervertebral disc structure and the viability of nucleus pulposus cells in isosmotic or hyperosmotic environment for at least 4 weeks.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R681.5;R-332

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