摘要
目的:观察周期性牵张应力对体外培养的人退变髓核细胞增殖的影响,为椎间盘退变的预防与治疗提供新的理论依据。方法:对体外培养的人退变髓核细胞利用甲苯胺蓝、番红O和Ⅱ型胶原免疫细胞化学染色进行鉴定,并将P3代细胞接种于弹性膜片上,待贴壁后移入ElectroForce 3200力学试验仪搭载的BioDynamic生物反应舱内对其施加0、5%、10%和15%的应变作用3h,受力完成后继续培养24h,用流式细胞仪测定细胞周期,计算S期百分数及增殖指数。结果:应力刺激后各组细胞贴壁及形态良好;应变作用3h后,0、5%、10%及15%应变组的S期百分数分别为1.86±1.22、2.07±0.09、7.56±1.40及7.55±3.25,增殖指数分别为3.47±2.52、7.88±2.45、16.05±1.51及14.7±0.61。5%应变组的增殖指数较0应变组明显增加(P<0.05);10%和15%应变组较0和5%应变组的S期百分数及增殖指数明显增加(P<0.05);10%和15%应变组间比较差异无显著性(P>0.05)。结论:在一定周期性应力范围内,髓核细胞的增殖能力增强。
Objectives： To investigate the effects of cyclic distraction stress on the proliferation of human nucleus pulposus cells（HNPCs）. Methods： HNPCs were cultured in vitro and identified via the toluidine blue, safranin O and immunocytochemical stain of collagen Ⅱ. P3 HNPCs were inoculated on elasticity polymer. After HNPCs were attached, BioDynamic test instrument was used to exert different distraction strain（0, 5%, 10%, and 15%） on HNPCs for 3 hours; after that, HNPCs were cultured for 24 hours, and the flow cytometry was used to examine the cell cycle and calculate the S phase fraction and proliferative index. Results： The cell adhesion and growth were well under different strain for 3h; S phase fraction in 0, 5%, 10% and 15% strain groups was 1.86＋1.22; 2.07＋0.09, 7.56＋1.40 and 7.55＋3.25 respectively. Proliferative index in these groups was 3.47＋2.52, 7.88＋2.45, 16.05＋1.51 and 14.7＋0.61 respectively. Compared with 0 strain group, pro- liferative index in 5% strain group increased significantly（P〈0.05）. Compared with 0 and 5% strain group, S phase fraction and proliferative index in 10% and 15% strain groups increased significantly （P〈0.05）; but S phase fraction and proliferative index showed no significant difference between 10% and 15% strain groups（P〉 0.05）. Conclusions： The proliferation of HNPCs increases under some stress.
出处
《中国脊柱脊髓杂志》
CSCD
北大核心
2012年第7期645-650,共6页
Chinese Journal of Spine and Spinal Cord
基金
国家自然科学基金青年基金项目(项目批准号:308011680)
天津市应用基础及前沿技术研究计划(项目编号:09JCZDJC19600)
天津市卫生局科研基金攻关项目(10KG111)
关键词
髓核细胞
周期性牵张应力
增殖
细胞周期
Cyclic distraction stress
Human nucleus pulposus
Proliferation
Cell cycle
