CAJ | 학술논문

目的研究周期性张应变对体外培养的后纵韧带骨化(OPLL)患者的后纵韧带细胞形态及凋亡的作用.方法组织块贴壁法培养后纵韧带细胞,选用第3代细胞,长至80%融合时,用含1%血清的培养基同步化24 h后,使用Flexerce114000TM细胞应变加载系统对细胞施加周期性张应变,作用6及24 h,牵拉幅度为10%.运用流式细胞仪对细胞凋亡水平进行检测,β-gal染色评价细胞衰老率.同时,检测细胞碱性磷酸酸(ALP)、Ⅰ型胶原(Col Ⅰ)和骨钙素(OC)mRNA的变化.结果与不加力的对照组相比,受力组细胞被拉长,并呈垂直作用力的方向取向排列;6 h组较对照组凋亡率略有升高,但差异无统计学意义;24 h组则明显升高(P＜0.05);6 h组与24 h组较对照组细胞衰老率无明显变化.ALP、Col Ⅰ和OC mRNA表达水平均升高,24 h组升高更明显(P＜0.05).结论周期性张应变对后纵韧带细胞的形态具有影响,应变刺激在促进OPLL细胞成骨分化的过程中,同时促进了细胞凋亡和功能的改变.
목적 연구주기성장응변대체외배양적후종인대골화(OPLL)환자적후종인대세포형태급조망적작용.방법 조직괴첩벽법배양후종인대세포,선용제3대세포,장지80%융합시,용함1%혈청적배양기동보화24 h후,사용Flexerce114000TM세포응변가재계통대세포시가주기성장응변,작용6급24 h,견랍폭도위10%.운용류식세포의대세포조망수평진행검측,β-gal염색평개세포쇠로솔.동시,검측세포감성린산산(ALP)、Ⅰ형효원(Col Ⅰ)화골개소(OC)mRNA적변화.결과 여불가력적대조조상비,수력조세포피랍장,병정수직작용력적방향취향배렬;6 h조교대조조조망솔략유승고,단차이무통계학의의;24 h조칙명현승고(P＜0.05);6 h조여24 h조교대조조세포쇠로솔무명현변화.ALP、Col Ⅰ화OC mRNA표체수평균승고,24 h조승고경명현(P＜0.05).결론 주기성장응변대후종인대세포적형태구유영향,응변자격재촉진OPLL세포성골분화적과정중,동시촉진료세포조망화공능적개변.
Objective To investigate the effect of mechanical stretch force on the morphologic and apoptotic changes of fibroblasts derived from the OPLL (ossification of posterior longitudinal ligaments)patients.Methods The third passage cells were collected and subjected to 10% elongations cyclic mechanical stretching for 6 h and 24 h with Flexerce114000TM strain unit.Their morphologic changes were observed and the post-stretching apoptotic rates quantified by flow cytometer.The gene expressions of alkaline phosphatase (ALP),collagen types Ⅰ (COL Ⅰ) and osteocalcin (OC) were examined.Results The treated cells were arranged along the vertical direction of force.Stretch force led to a slight increase of apoptosis rate at 6 h and a significant increase of apoptosis rate at 24 h.No significant difference in cellular senescence was observed between control group and treated group.The mRNA expressions of ALP,COL Ⅰand OC were positively up-regulated by cyclic stretch at 24 h.Conclusion Stretching force can affect the cellular morphology,promote the osteogenic differentiation and enhance the cellular apoptosis.