SKIN PROTECTION 113 Results


Immunostaining of cells treated with LED- BL (45 J/cm2 and 5 J/cm2


for 1.5 hours for keratinocytes for 15 minutes for fibroblasts)


confirmed that blue light causes DNA damage, which was visualised with an increase of PCNA and 8-OHdG immunolabelling. Olive fruit extract produced a significant


reduction of DNA damage induced by LED- BL exposure: the immunofluorescence of the antibody against PCNA decreased, whereas the immunofluorescence of the antibody against 8-OHdG completely disappeared.


These data, in line with those produced


by Western Blot analysis, corroborated the efficacy of olive fruit extract towards blue light-induced DNA damage.


Effect of olive fruit extracton modulation of MMP-1, MMP-12 and collagen type I in LED-BL treated fibroblasts


Design The expression of matrix metalloproteinases (MMP-1, MMP-12) and collagen type I was quantified through specific antibodies, which were detected using a chemiluminescent system. The signal intensity of the bands was expressed


Control


Blue light 10 µg/mL 25 µg/mL 50 µg/mL Olea-HT10


100 µg/mL b Control Blue light 10 µg/mL


25 µg/mL Olea-HT10


50 µg/mL a


Figure 6: Immunofluorescence of keratinocytes (a) and fibroblasts (b) treated with LED-BL and olive fruit extract and stained with antibody against PCNA (green). The nuclei were counterstained with 4′,6- diamidino-2-phenylindole (DAPI) (blue).


as arbitrary densitometric unit (ADU) using, as reference, the density of the α- tubulin bands.


Results Western Blot analysis, performed on fibroblasts exposed at LED-BL (5 J/cm2 has shown high levels of MMP-1


),


(collagenase) and MMP-12 (elastase) and the complete absence of collagen type I. The pre-treatment with olive fruit extract reduces the expression of these matrix metalloproteinases and promotes the synthesis of collagen type I (Fig 8a, b). Interestingly, olive fruit extract not only counteracts blue light-induced degradation


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