CAJ | 학술논문

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。
배경：피부조직소처적역학배경이급상피세포적포전상태여상구유합화반흔형성과정관계밀절。목적：분석포외역학자격대세포포전적작용，병검측세포적증식상태진일보분석포전형태대세포증식등생리활동적영향。방법：통과FX-4000유성기저가재계통대인영생화각질형성세포(HaCaT)진행주기성랍신응력정현파가재，가재정식0.2 Hz，10%랍신폭도。재0，24，48 h대세포적포전형태진행대비，이용류식세포의검측분석세포적증식，병이용면역형광염색방법대비분석뉴단백재세포내적분포。결과여결론：HaCaT세포재가재24 h후분렬기세포교다，포전형태무명현변화；가재48 h후HaCaT세포형태발생교위명현적변화，분렬기세포교정태대조조략유감소；랍신응력작용하뉴단백적분포유세포핵주위적막구역향세포변연집중。결과표명：괄도적역학가재능구재보지세포포전화점부상태하，촉진세포증식；주기성지속랍신응력적작용시간시HaCaT세포포전형태화여기저점부위점적중요영향인소。
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 adhesion regions of immortalized human keratinocyte.