体外培养保存关节软骨组织细胞凋亡的研究

Chondrocyte apoptosis of culture-storaged articular cartilage in vitro

[目的]体外培养保存关节软骨组织,研究分析软骨细胞凋亡规律、培养液中组织代谢产物(NO、MDA、SOD)与细胞凋亡之间的相关性。[方法]切取新西兰大白兔膝关节骨软骨柱,使用普通无菌MEME培养液保存,分时间点检测实验指标:采用流式细胞技术测定细胞凋亡数量,比色法测定细胞培养液上清中一氧化氮(NO)、丙二醛(MDA)含量,以及超氧化物歧化酶(SOD)活性。[结果]随保存时间延长,软骨细胞凋亡率、培养液中NO、MDA含量均逐渐增加、SOD活力逐渐降低,与前段时间点相比较有统计学意义(P<0.05);培养液中代谢产物含量与细胞凋亡率高度相关(P<0.01)。[结论]体外培养保存软骨组织,细胞凋亡率逐渐增加,呈时间依赖性,28 d时尤为明显;NO诱导的细胞凋亡对保存软骨组织活性降低起到重要作用。

[ Objective] To research the features of chondrocyte apoptosis in vitro culture preserved articular cartilage, and correlation between apoptosis and culture metabolins, including nitric oxide （NO）, malondialdehyde （MDA）, and superoxide dismutase （SOD）. [ Method ] Osteochondral plugs, subsequently stored in culture medium of MEME, were harvested from the knee of New Zealand white rabbits. Chondrocyte apoptosis of the sample was measured by flow cytometry, and the contents of NO, MDA and SOD in culture solution were assayed by colorimetric technique at different time points. [ Result] The chondrocyte apoptosis and contents of the NO, MDA increased as the culture time elapsed, conversely, the SOD gradually became less active. There were statistical differences at each interval（ P 〈 0.05 ）. The content of culture metabolin had highly correlation with chon- drocyte apoptosis. （P 〈 0.01 ）. [ Conclusion] The results suggest that chondrocyte apoptosis gradually increased in vitro culture stored, especially at 28 days . Nitric oxide-induced apoptosis had a significant influence on tissue viability in articular cartilage.