周期性拉伸应力与人永生化角质形成细胞的黏附和铺展  

Cyclic mechanical stretch influences cell adhesion and spreading of immortalized human keratinocytes

作　　者：刘琨[1] 安美文[1] 王立[1] 黄晶晶[1] 

机构地区：[1]太原理工大学,山西省太原市030024

出　　处：《中国组织工程研究》2015年第1期119-123,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金支持(10672114);山西省自然科学基金支持(2007011011)~~

摘　　要：背景:皮肤组织所处的力学环境以及上皮细胞的铺展状态与伤口愈合和瘢痕形成过程关系密切。目的:分析胞外力学刺激对细胞铺展的作用,并检测细胞的增殖状态进一步分析铺展形态对细胞增殖等生理活动的影响。方法:通过FX-4000柔性基底加载系统对人永生化角质形成细胞(HaCaT)进行周期性拉伸应力正弦波加载,加载程式0.2 Hz,10%拉伸幅度。在0,24,48 h对细胞的铺展形态进行对比,利用流式细胞仪检测分析细胞的增殖,并利用免疫荧光染色方法对比分析纽蛋白在细胞内的分布。结果与结论:HaCaT细胞在加载24 h后分裂期细胞较多,铺展形态无明显变化;加载48 h后HaCaT细胞形态发生较为明显的变化,分裂期细胞较静态对照组略有减少;拉伸应力作用下纽蛋白的分布由细胞核周围的膜区域向细胞边缘集中。结果表明:适度的力学加载能够在保持细胞铺展和黏附状态下,促进细胞增殖;周期性持续拉伸应力的作用时间是HaCaT细胞铺展形态和与基底黏附位点的重要影响因素。BACKGROUND：The mechanical environment of skin tissue and spreading state of epithelial cels are closely related with the wound healing and scar formation process. OBJECTIVE：To analyze the effect of extracelular mechanical stimulation on cel spreading, to test the cel proliferation in order to analyze the effect of spreading form on cel proliferation and other physiological activities. METHODS： Cyclic sine wave mechanical stretching was exerted on immortalized human keratinocyte by using FX-4000 flexible substrate loading system, on the condition of 0.2 Hz and at frequency of 10% amplitudes. The spreading form was compared at 0, 24 and 48 hours, the cel proliferation was analyzed with flow cytometry, and the distribution of vinculin was analyzed with immunofluorescence staining. RESULTS AND CONCLUSION： human keratinocyte would keep the spreading state and could induce more cel proliferation by 24 hours mechanical stretching stimulation. Conversely, after stimulated for 48 hours, the morphology of the human keratinocyte was significantly changed, and the number of human keratinocyte in the division stage was larger than that in the static control group; under tensile stress, the distribution of vinculin was transformed from the surrounding nucleus membrane area to the cel edge. The results indicate that proper mechanical stimulate can increase cel proliferation with keeping cel spreading and adhesion state; the stimulating time of continuous cyclic stretching is the major factor to determine cel spreading morphology and＆amp;nbsp;adhesion regions of immortalized human keratinocyte.

关 键 词：生物力学 应力 物理 细胞粘附 细胞增殖 干细胞 培养 周期性拉伸 应力 人永生化角质形成细胞 HACAT 铺展 纽蛋白 增殖 黏附 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]