One of an essential characteristic of human skin are time dependent mechanical properties. Here, we demonstrate that stiffness of human dermal fibroblast correlates with age and it can be restored after anti-wrinkle tripeptide treatment. The stiffness of human fibroblasts isolated from donors of 30-, 40- and 60 years old were examined. Additionally the effect of anti- wrinkle tripeptide of latter cells was investigated. The atomic force microscopy measurements were performed on untreated fibroblast as well as on treated with the peptide. The Young’s modulus for two indentation depths 200 and 600 nm of each cell type was determined. The Young’s modulus increases with age of the cells. The highest values of Young’s modulus were obtained for fibroblasts collected from 60 years old donors, for indentation depth of ~200 nm. For larger indentation depth of 600 nm there are no significant differences in stiffness between cells. Fibroblasts treated with the anti-wrinkle tripeptide exhibit lower Young’s modulus. The cells derived from 40- and 60-years old donors restored stiffness characteristic to the level of 30 years old subjects. The results show correlation between stiffness and age of the human fibroblast as well as impact of anti-wrinkle tripeptide on the mechanical properties of skin cells.

A b s t r a c tIntroduction: Fibroblast growth factor 1 (FGF-1) is a powerful mitogen involved in the stimulation of DNA synthesis and the proliferation of a wide variety of cell types. Fibroblast growth factor 1 was genetically modified to improve its thermal stability and resistance to protease degradation without losing its biological activity. Aim: To study the impact of Q40P/S47I/H93G rFGF-1 on skin cells, its penetration through the skin and the evaluation of the rFGF-1-cosmetic product properties. Material and methods:In vitro studies included the examination of primary fibroblast and keratinocyte viability after the incubation with rFGF-1. The penetration abilities of rFGF-1 in various formulations and carrier systems were examined ex vivo by the Raman spectroscopy. In vivo studies -HF Ultrasound and 3D Imaging System -were used to evaluate the anti-aging properties of creams containing rFGF-1. Results:In vitro studies demonstrated that rFGF-1 strongly enhanced the viability of the treated cells. The Raman Spectroscopy analysis indicated that rFGF-1 encapsulated in lipid spheres penetrate through the stratum corneum to the depth of 60 µm, and added to the o/w formulation -could penetrate to a depth of 90 µm. The results obtained from Primos revealed the reduction of the volume and the depth of the wrinkles. Changes in the skin structure in the analyzed areas were evaluated by HF Ultrasonography. Conclusions: Recombinant FGF-1 strongly stimulated fibroblast and keratinocyte proliferation. However, the transition of this protein through the SC required an appropriate carrier system -lipid spheres. All tests -in vitro, ex vivo and in vivo -have proved that rFGF-1 is a substance with a potentially wide spectrum of use.

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Dermocosmetics for people with hyperpigmentation are an essential part of their medical treatment. In case of epidermal hyperpigmentation, the recipe of individually chosen and tested combination of ingredients enables us to reach satisfactory results.

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