摘要
目的研制一种体外细胞单轴循环牵伸模型。方法用硅树脂材料制备带微沟槽培养皿,运用有限元软件ABAQUS分析培养皿应变分布。将肌腱干细胞(tendon stem cells,TSCs)在培养皿中培养,观察TSCs贴壁生长情况。可编程控制器(programmable logic controller,PLC)控制牵伸模型机械运行部分对培养皿周期性牵伸。按不同牵伸频率(0.5、1.0、2.0、3.0 Hz)及应变(4%、8%、16%、20%)分为16组连续牵伸24 h,检测培养皿实际运行次数及位移量变化;倒置显微镜观察TSCs有无脱落及漂浮;在牵伸时间为8、12 h(频率1 Hz、应变8%)时,用激光共聚焦扫描显微镜观察细胞骨架中F-actin形态变化。结果硅树脂培养皿应变分布较均匀,培养皿底膜有效应变范围约占总面积的91%。TSCs在培养皿中培养10 h时,全部贴壁且沿微沟槽方向生长。在不同牵伸条件下牵伸模型运行稳定,硅树脂培养皿实际运行频率、应变位移距离和设定频率及理论应变位移距离比较差异无统计学意义(P>0.05);TSCs无明显脱落及漂浮;牵伸组F-actin束变细,部分发生断裂,红色荧光(F-actin)染色减弱且分布不均匀。结论体外细胞单轴循环牵伸载荷模型建模成功。该模型操作简便、参数设计范围宽,可模拟不同条件进行体外贴壁细胞生物力学载荷实验研究。
Objective To design a uniaxial cyclic stretching model of tendon stem cells （TSCs） in vitro. Methods After our silicone-made micro-grooved dishes were analyzed for strain distributions by the finite element software ABAQUS. Freshly isolated and identified TSCs were cultured on the dishes, and then observed for their adherent growth. A programmable logic controller （PLC） was used to make the mechanical running parts of the model do cyclically stretching for the dishes. Dishes were divided into 16 groups, with the cyclic stretching for 24 h under different stretch frequencies （0.5, 1.0, 2.0 and 3.0 Hz） and strains （4%, 8%, 16% and 20%）. The actual running frequency and displacement distance of these silicone dishes were measured. Inverted microscopy was employed to observe the falling off or floating of the TSCs. F-actin in the cytoskeleton were observed by laser confocal scanning microscopy when the cells were stretched for 8 or 12 h under the frequency of 1 Hz and strain of 8%. Results The strain distribution was uniform on the self-made silicone dishes. Effective strain range of the bottom membrane accounted for about 91% of its total area in the dishes. When cultured on the dishes for 10 h, all TSCs attached on the dishes and grew along with microgroove direction. Stretching model ran stably under different stretching conditions. There was no significant difference in the actual running frequency and strain displacement distance of the silicone dishes with the set frequency and theoretical strain displacement distance （P〉0.05）. No obvious falling off or floating was observed in the TSCs cultured on the dishes. F-actin filaments seemed much thinner, and some of them were broken, and the red fluorescence dyeing （F-actin） was weakened and unevenly distributed in the streched cells than the cells without stretching. Conclusion Our established uniaxial cyclic stretching model is easy to operate, with wild range of parameters, and is used to simulate conditions for adherent cells under diff
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第11期1142-1146,共5页
Journal of Third Military Medical University
基金
国家自然科学基金(81230040)~~
关键词
肌腱干细胞
微沟槽
牵伸
三维有限元
肌腱病
eywords： tendon stem cells
microgroove
stretching
three dimensional finite element
tendinopathy
