中文摘要：

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，探讨渗透压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（300mOsm／kg，等渗组）或高渗（410mOsm／kg，高渗组）培养基中进行整体器官培养，存培养前和培养后第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（P〈0．05），第7、21和28天分别为10504±710、10860±711、10713±953，与培养前相比较无显著性差异（P〉0．05）；等渗组第7、14、21、28天分别为11350±351、11207±385、10914±300、10862±229，与培养前比较无显著性差异（P〉0．05）：两组在第7、21和28天时无显著性意义（P〉0．05）。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第7天高渗组和等渗组分别为3．33±0．28mg／100mg和2．83±0．25mg／100mg，均较培养前（5．03±0．37mg/100mg）明显降低（P〈0．01），第14、21、28天时与第7天相比下降不明显（P〉0．05），两组相同时间点的差异无显著性意义（P〉0．05）。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。

英文摘要：

Objective：To develop an organ culture model of rabbit intervertebral disc in vitro and investigate effects of osmotic loading on cell viability and metabolism of the discs.Method：Thoracolumbar and lumbar intervertebral discs were harvested immediately after sacrifice from 10 New Zealand white rabbits （4-6 months old,n=9/animal） ,which were divided into two groups.The intervertebral discs were maintained in organ culture with iso-osmotic （300mOsm/kg,iso-osmotic group） or hyperosmotic （410mOsm/kg,hyperosmotic group） media. Cell viability,structural integrity and proteoglycan content were assessed using Mitotracker Green probe,histochemical and biochemical methods before culture and on 7,14,21,28 days after culture.Result：The fluorescence intensity of fresh intervertebral disc was the maximum（11503±402） before the culture.During organ culture in vitro,there was a signicant decrease in fluorescence in the hyperosmotie group on 14 days （9202± 907）（P〈0.05） compared to that prior to culture,the fluorescence intensity of hyperosmotic group on 7,21 and 28 days were 10504±710,10860±711 and 10713±953 respectively.The fluorescence yield on 7,14,21 and 28 days of iso-osmotic group were 11350±351,11207±385,10914±300 and 10862±229,and there was no signicant difference compared to fresh intervertebral disc （P〉0.05）.There was no signicant change in fluorescence yield on 7,21 and 28 days between two groups（P〉0.05）.In the culture,the nucleus pulposus and anulus brosus maintained the structural characteristics of the intact tissue on the whole.There was a signicant decrease of proteoglycans in both hyperosmotic group（3.33±0.28） and iso-osmotic group（2.83±0.25） on 7 days culture time compared to fresh intervertebral disc （5.03±0.37）（P〈0.01）.There was no additional loss of proteoglycans was detected on 14,21 and 28 days than on day 7 （P〉0.05）.No signicant difference in proteoglycan content was found between tow groups on the same period （P〉0.05）.Cone