MARINE INGREDIENTS
exopolysaccharide, thanks to its stimulating effect on elastin synthesis by fibroblasts, aims to improve skin elasticity.
Effect on collagen I synthesis For the collagen immunostaining in 2D, NHDF were isolated from healthy skin obtained from plastic surgery and used in passage 7, cultured in monolayer on Labtek at a density of 7000 cells by well. NHDF were treated with 0.02% of the marine exopolysaccharide dissolved in DMEM Glutamax medium supplemented with 2% fetal bovine serum and 1% penicillin/streptomycin, or untreated. The cells were cultivated for 120 hours and each experimental condition was performed in triplicate.
In the absence of product, the staining of collagen I fibres is low. In the presence of the marine exopolysaccharide, there was a clear increase of collagen I (see Fig. 6). Treatment with image analysis was performed on the photographs. The colour intensity is measured by image analysis software and related to 100 cells. The results are shown in Figure 7. Treatment with 0.02% of the marine exopolysaccharide increases by 20% the synthesis of collagen I by human fibroblasts in monolayer. These results suggest that the marine exopolysaccharide thanks to its stimulating effect on collagen I synthesis by fibroblasts, aims to improve skin firmness.
Affinity for the skin Human skin explants of 8 mm of diameter were treated with the marine exopolysaccharide for 1 hour. At the end of the incubation period, the samples were freeze-dried to fix and dehydrate the skin. The samples were observed with a JEOL JSM 6301F SEM (field effect scanning electron microscope).
The photographs show scanning electron microscope observations of the surface of an epidermis. Furrows and variations in cutaneous relief can clearly be seen (Fig. 8). After treatment with the marine exopolysaccharide, the furrows are filled in and the differences in relief are smoothed out and made uniform.
Immediate lifting effect on cutaneous relief (in vivo Test 2)
Smoothing effect was evaluated on four Caucasian female volunteers, aged between 45 and 65 years, applying once a cream containing 2% of the marine exopolysaccharide (i.e. 0.02% final concentration of the pure exopolysaccharide) to crow’s feet selected at random. Evaluation was made by measuring the skin’s roughness and the cutaneous relief dispersion using fringe projection
a b
Figure 6: Immunostaining of collagen I fibres produced by human fibroblasts a) untreated or b) treated with 0.02% of EPS Seafill.
14.0 13.5 13.0 12.5 12.0 11.5 11.0 10.5 10.0
Non-treated fibroblasts
15 minutes after application as shown in Figure 9.
+20% Fibroblasts
treated with 0.02% of EPS Seafill
Figure 7: Quantification of the collagen I staining intensity in human fibroblasts non treated or treated with EPS Seafill.
before and 15 minutes after the product application.
Fifteen minutes after applying the cream containing 2% of the marine exopolysaccharide, a reduction in average roughness of –8.7% on average and up to –17.5% is observed as well as a reduction in roughness with regard to the average quadratic variation of –9.9% on average and up to –18%.
After only one application of a cream containing 2% of the marine exopolysaccharide on crow’s feet wrinkles we showed a roughness smoothing effect as well as a cutaneous relief lifting effect
a
Immediate anti-wrinkle effect (in vivo Test 2) Anti-wrinkle efficacy was evaluated on 17 Caucasian female volunteers, aged between 45 and 65 years, applying once a cream containing 2% of the marine exopolysaccharide to facial crow’s feet selected at random versus a base cream alone (placebo). Evaluation was made by measuring the depth of the crow’s feet wrinkles using fringe projection and with illustrative photographs before application and 15 minutes, 3 hours and 6 hours after the product application. A unique application of a cream containing a concentration of 2% the marine exopolysaccharide reduces the depth of the main wrinkle by –5.1% on average and up to –16% after 15 minutes (significant variation), –1.9% on average and up to –15% after 3 hours, –0.1% on average and up to –20% after 6 hours (see Fig. 10). This effect is visible as shown in Figure 11.
In comparison, application of the placebo cream reduces the depth of the main wrinkle by only –0.6% on average and up to –15%, after 15 minutes (significant variation), and –0.2% on average and up to –24% after three hours. After six hours, the control cream has a tendency to increase the depth of the wrinkle. Thus the marine exopolysaccharide
b
Figure 8: SEM observation of human skin explants a) treated or b) not with EPS Seafill (enlargement x100).
April 2012 PERSONAL CARE 49
Collagen I/100 cells (%)
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