Development of a new photo-aging model system: A quick evaluation method for photo-damage utilizing elastin-promoter/green fluorescence protein (GFP) reporter gene transfected dermal fibroblasts

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For the purpose of prevention of photo-aging such as deep wrinkle formation, a great number of sunscreens against broad-band spectrum of solar ultraviolet (UV) has been produced. However, in vivo evaluation of sunscreens for anti-photoaging requires long time. In the present experiments, a new evaluation system have been developed. Cultured mouse fibroblasts (3T3) were transfected with human-elastin promoter linked to green fluorescence protein (GFP) reporter gene. After exposure to UVB, green fluorescence intensity of GFP was observed. In addition, after injection of the cells into the mouse skin, fluorescence intensity was quantified using spectrofluorometer with fiber optics (the SPEX FluoroMax-3 of Jobin Yvon Inc.). The cultured cells expressed green fluorescence in UVB-dose dependent manner. The fluorescence of injected cells was measurable from skin surface. These results indicate that quantitative measurement of fluorescence intensity of injected cells to the mice skin might be useful for early evaluation of solar elastosis formation and protective effect of sunscreens.