MARINE INGREDIENTS
■ Placebo cream ■ Cream containing 0.01% pseudoalteromonas expolysaccharides
6 4 2 0
–2 –4 –6 –8
–10 –12
–11.1% 2 hours 8 hours 20 hours Figure 4: Maximum roughness reduction variations in percentage.
Instrumental evaluation of skin hydration
A panel of 20 female volunteers (mean age 44.3) applied a placebo cream on the right side of the face and a cream with 0.1% pseudoalteromonas exopolysaccharides on the left side for 20 days twice a day. Measures of skin hydration were performed with a Corneometer CM 825 at the beginning, 2 hours and 8 hours after the first application and at the end of the treatment. The basal control instrumental measurements were performed on the cheeks.
The values were compared by means of Repeated Measures Analysis of Variance and Bonferroni Test. The differences between the data groups were considered significant when the probability p was 0.05.
The cream containing 0.1%
pseudoalteromonas exopolysaccharides provided a statistically significant increase in the mean basal values of skin hydration after 2 hours, 8 hours, and 20 days. The statistical comparison with the placebo showed a significant difference.
Instrumental evaluation of skin roughness
A panel of 20 female volunteers (mean age 41 years) applied a placebo cream on the right side of the face and a cream with 0.01 % pseudoalteromonas exopolysaccharides on the left side for 20 days twice a day. Skin replicas were taken at the beginning, 2 hours and 8 hours after the first application and at the end of the treatment. The skin imprints were analysed by designed image processing software (Quantilines, Monaderm). The basal control instrumental measurements were performed on the cheeks. The values were compared by means of Repeated Measures Analysis of Variance
–8.4% –9.3%
35 30 25 20 15 10 5 0
Control
34.0%
Pseudoalteromonas ferment extract 0.1 mg/mL
1 mg/mL
Figure 5: Increase of type I collagen synthesis induced by pseudoalteromonas ferment extract after 48 hours.
and Bonferroni Test. The differences between the data groups were considered significant when the probability p was ≤0.05. The cream containing 0.01 % pseudoalteromonas exopolysaccharides showed a statistically significant decrease in maximum roughness values on all the cheeks. The statistical comparison with the placebo showed a significant difference after 2 and 8 hours, and at the end of the 20 days of treatment.
Cryoprotective profile By frosting and defrosting carboxyfluorescein (CF) liposomes in presence of 1% pseudoalteromonas ferment extract, it is possible to evaluate its cryoprotective activity on the lipidic bilayers.
Liposomes containing high concentrations of CF were prepared. When the instability of the bilayers caused the release of the encapsulated CF of the liposome, and its consequent dilution in the medium, an increased fluorescence was detected.
The defrosted CF liposomes incubated
with the glycoprotein showed a fluorescent intensity very close to that of the non- frozen liposomes, meaning that the liposomes structure had suffered little damage during the freezing process.
Cryoprotective activity assay by Differential Scanning Calorimetry (DSC) Differential Scanning Calorimetry (DSC) measurements were performed with DSC821 STRAe SYSTEM apparatus. Solutions of 0.06%, 0.6%, 2.4%, 5% and 10% (w/v) pseudoalteromonas ferment extract were prepared in pure water. Solutions of 2.5%, 5% and 10% of Bovine Serum Albumin (BSA) in pure water were used as controls. Samples were put into capsules, transferred to the calorimeter,
cooled up to –40˚C at the rate of 5˚C/min and kept for 10 minutes at that temperature. Then, they were heated up to 10˚C at a rate of 5˚C/min and the melting behaviour of the water was registered. Pseudoalteromonas ferment extract induced a depression in the melting point in a dose-response manner, reaching a decrease of almost 2ºC at a concentration of 10%. Albumin samples only decreased the melting point by 0.5˚C at the same concentration. The glycoprotein has a specific activity on delaying the ice formation in pure water, and, therefore, it is expected to present cryoprotective properties, protecting skin at low temperatures.
Effect on cell adhesion in human dermal fibroblasts It was studied the efficacy of pseudoalteromonas ferment extract in inducing cell adhesion on human dermal fibroblasts (HDF) by Crystal Violet Dye Elution (CVDE) method.
The substrates were coated with 1 ng/mL and 1 mg/mL pseudoalteromonas ferment extract on a support, the cells were added, and a wash was performed. Substrate was coated with 20 µg/mL of type I collagen was used as control of cellular substrate adhesion.
HDF cells showed an important increase in cellular adhesion when grown on 1 mg/mL coated pseudoalteromonas ferment extract substrate, enhancing it by 125% after 5 hours. At 1 ng/mL, the levels of bioadhesion were comparable to those for 20 µg/mL type I collagen.
Effect on cell growth in human keratinocytes It was studied the efficacy of pseudoalteromonas ferment extract in inducing cell growth on human keratinocytes (HEK) by CVDE method.
April 2012 PERSONAL CARE 55
Maximum roughness reduction variations (%)
Increase of type I collagen synthesis (%)
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