ACTIVES
profilometric measurements were performed in the area of the outer corner of the eye using the PRIMOS Pico system. Figure 6 shows the reduction of wrinkle depth in the region of the lateral corner of the eye at the starting point (D0), after 28 days (D28) and 56 days (D56) of treatment with a cream formulation containing 1% PGFE/SAP. After 28 days of treatment a reduction
in wrinkle depth could be observed in 90% of the test subjects. The maximum reduction in wrinkle depth measured was 25.5% with an average value of 9.1% after 28 days. After 56 days of treatment, a significant reduction of wrinkle depth was observed in 100% of test subjects. The average reduction in wrinkle depth came to 15.8% and the maximum reduction to 46.9%.
Visual evaluation (VisioFace Quick) In both Study 1 and Study 2, high- resolution photos of the face were taken. In order to obtain standardised images in terms of brightness, image size and field of vision, a VisioFace Quick device was used. The pictures in Figure 8 show an example from Study 1 and Figure 9 an example from Study 2. Figure 8 shows photographs of the
crow’s feet area at the start (D0) and the end (D56) of Study 1. A clearly visible decrease in the appearance of lines can be observed. Figure 9 shows pictures of the forehead region at measurement times D0, D28 and D56 taken in Study 2. The frown line/forehead wrinkle well recognisable in the centre of the photograph was clearly reduced by treatment with a cream formulation containing 1% PGFE/SAP. In addition an overall smoothing of the forehead area can be seen.
Discussion The results discussed in this paper demonstrate the safety and effectiveness of the combination of pomegranate flower extract and sodium ascorbyl phosphate. PGFE/SAP exhibited the desired effects in a series of in vitro and in vivo experiments. MMP-1 gene expression analysis in
primary dermal fibroblasts (Fig. 1) showed an increase in MMP-1 mRNA gene expression evoked by UVA irradiation. This effect is described in literature5
and
therefore was expected and reproducible. The increased MMP-1 mRNA gene expression in irradiated cells was not only halted but actually reduced by 45% after treatment with pomegranate flower extract. It is worth noting that PGFE also had an effect on endogenous MMP-1 mRNA gene expression, since there was also a 58% reduction in MMP-1 gene expression in PGFE treated non-irradiated cells. Thus
50 PERSONAL CARE March 2012 D0 D28
effect in the second in vivo study. The 28- day application of a cream formulation containing 1% PGFE/SAP produced a 9.1% reduction in line depth in the area around the lateral corner of the eye (Fig. 6). After 58 days of application, line depth decreased by 15.8%. An example from this in vivo study is given in Figure 9. The different in vivo and in vitro tests
D56
Figure 9: High-resolution photographs of the forehead region.
PGFE appeared to be able to influence not only the extrinsic photoageing process evoked by UVA irradiation but also chronological skin ageing in terms of lowering MMP-1 expression. In order to demonstrate this effect at
protein level, an ELISA quantification of MMP-1 in aged dermal fibroblasts was carried out (Fig. 4). Furthermore it had to be clarified whether pomegranate flower extract activity was still present in the PGFE/SAP after freeze drying. In all the samples containing PGFE MMP-1 synthesis was decreased on average by 20%. This showed that PGFE’s properties for inhibiting MMP-1 synthesis had been retained in the freeze drying process and the planned standardisation of PGFE/SAP as regards its activity was successful. The reproducibility of the results after six months showed that PGFE/SAP remained stable and the inhibiting activity in terms of MMP-1 synthesis had not decreased. The impairment of cell vitality in the
UV cytotoxicity test from repeated UVA irradiation was diminished from 41% to 12% by using 0.1% pomegranate flower extract (Fig. 2). This showed that PGFE has cell-protecting properties with respect to UVA-induced impairment of cell vitality. The anti-ageing effect of PGFE/SAP
demonstrated in the in vitro tests was confirmed by the in vivo studies carried out. The use of a cream formulation containing 1% PGFE/SAP in Study 1 increased collagen thickness significantly (by 32.5%) on the volar forearm (Fig. 5 and Fig. 7) and visibly reduced the appearance of wrinkles around the lateral corner of the eye (Fig. 8). PGFE/SAP also showed an anti-ageing
demonstrated that PGFE/SAP was able to meet the desired requirements in terms of its activity. It was possible to demonstrate MMP-1 inhibition from the PGFE component in the ELISA experiment and the expected increase in collagen synthesis from the SAP component with the help of the ultrasonography results. The results of both effects were visible with profilometric-determined (Fig. 6) and visually-assessed (Fig. 8 and Fig. 9) reduction in line depth. The tests conducted for allergic potential, mutagenicity, phototoxicity as well as skin and eye irritation demonstrated the safety of use of PGFE/SAP. Together with the in vitro and in vivo data collected on efficacy the result is therefore an anti-ageing product that is very effective and safe to use.
PGFE/SAP is sold under the Cosmetochem trade name Colla-Gain™
References 1 Pugliese P. Physiology of the skin II. Allured (2001)
2 Rittie L, Fisher GJ. UV-light-induced signal cascades and skin aging. Ageing Res Rev 2002; 1 (4):705-20.
3 Kahari VM, Saarialho-Kere U. Matrix metalloproteinases in skin. Exp Dermatol 1997; 6 (5): 199-213.
4 Helfrich YR, Sachs DL, Voorhees JJ. Overview of skin aging and photoaging. Dermatol Nurs 2008; 20 (3): 177-83.
5 Puizina-Ivic N. Skin aging. Acta Dermatovenerol Alp Panonica Adriat 2008; 17 (2): 47-54.
6 Yaar M. Skin aging: observations at the cellular and molecular levels. Isr J Med Sci 1996; 32 (11): 1053-8.
7 Yaar M, Eller MS, Gilchrest BA. Fifty years of skin aging. J Investig Dermatol Symp Proc 2002; 7 (1): 51-8.
8 Callaghan TM, Wilhelm KP. A review of ageing and an examination of clinical methods in the assessment of ageing skin. Part I: Cellular and molecular perspectives of skin ageing. Int J Cosmet Sci 2008; 30 (5): 313-22.
9 BASF. Sodium Ascorbyl Phosphate. 2005; July MEMC 050423e-00
10 Nusgens BV, Humbert P, Rougier et al. Topically applied vitamin C enhances the mRNA level of collagens I and III, their processing enzymes and tissue inhibitor of matrix metalloproteinase 1 in the human dermis. J Invest Dermatol 2001; 116 (6): 853-9.
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