Cyclic mechanical stretch influences cell adhesion and spreading of immortalized human keratinocytes / 中国组织工程研究

ABSTRACT

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.