周期性牵张应力对人退变髓核细胞增殖的影响

Effects of cyclic distraction stress on proliferation of human nucleus pulposus cells

中文关键词:  髓核细胞  周期性牵张应力  增殖  细胞周期

英文关键词:Cyclic distraction stress  Human nucleus pulposus  Proliferation  Cell cycle

中文摘要:

  【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。

英文摘要:

  【Abstract】 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 collagenⅡ. 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, proliferative 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.