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As skin loses its elasticity and muscles weaken through age, loose skin can accumulate around the eyes, forming folds in the eyelids. Fat, which cushions the eyes in their sockets moves forward out of the ocular cavities and accumulates in bulging bags around the eyelids.
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Baggy eyelids are known medically as dermatochalasis and are commonly improved by performing a blepharoplasty, a surgical procedure which involves an inner or outer incision of the eyelid to extract the fat and excess skin. Blepharoplasty is the most common aesthetic procedure performed by plastic surgeons in America [Castro, E, Foster, JA (1999) Upper lid blepharoplasty. Facial Plast. Surg.15 (3): 173 ].
The other major reason for puffy eyes is water accumulation, known as eyelid oedema. Fluid may build up for several reasons, two of the major reasons being poor lymphatic circulation and increased capillary permeability.
Eyeseryl is a tetra peptide with anti-oedema properties with a proven efficacy in reducing puffy eye bags.
o -The draining effect of Eyeseryl achieves a reduction of puffy eye bags in only 15 days – fast anti-eye bag action
Eyeseryl has been tested in vitro for ocular irritation and is non-irritant at the concentration of use. All the raw materials involved in the formulation are regarded as safe at the concentrations of use.
Glycation is a non enzymatic reaction between glucose or another monosaccharide and a protein. The aldehyde group of the monosaccharide reacts with the terminal amino groups of the amines to produce a Schiff’s base, forming fructosamine, or Amadori adduct.
At a second stage, the Amadori compound may react producing a group of irreversible reactions (oxidation, crosslinking...).
Collagen cross-linking is one of the main causes of the formation of eyebags. This test was carried out to evaluate the ability of EYESERYL ® to inhibit glycation.
It is known that some enzymes can suffer glycation in vivo. Among those enzymes we can find the Cu, Zn-Superoxide Dismutase (SOD). The SOD is an enzyme that converts superoxide radicals to hydrogen peroxide and oxygen.
The incubation of SOD with glucose or other monosaccharides gives rise to glycation, which inactivates the enzyme.
Some compounds can inhibit SOD glycation and, therefore, maintain its activity.
In this study, the inactivation of SOD by its reaction with fructose is used as a model of glycation. The effect of Eyeseryl as an inhibitor of glycation is evaluated. A method to assess the SOD activity by the inhibition of the transformation of xanthine to uric acid with the enzyme xanthine oxidase is used. With this reaction, the WST-1 (2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) is transformed to formazan, a compound which absorbs at 470nm.
If SOD is added to this reaction, radical O2-. is captured and the formation of this colored compound is avoided.
Results
The results show there is and increase in the SOD activity, which means that Eyeseryl inhibits its glycation.
The endothelial cell lining of the internal vasculature defines a semi-permeable barrier between the blood and the interstitial spaces of the body.
Endothelial cell monolayers cultured on semi-permeable membranes have been shown to form adherent tight junctions. The in vitro vascular permeability Assay Kit provides an efficient system for evaluating the effects of chemicals and drugs on endothelial cell adsorption, transport and permeability.
A multitude of vasoactive cytokines, growth factors, and signal modulators react with endothelial cell substructure components to control permeability. VEGF, Interleukin alpha and beta, TNF alpha, and IFN gamma have been shown to increase endothelial monolayer permeability.
The test is performed in a 24-well tissue culture plate with 12 cell culture inserts. The inserts contain 1.0 µm symmetrical pores within a transparent polyethylene membrane. The high pore density membranes permit amplified rates of basolateral diffusion with molecules of interest for permeability assays. The membrane is tissue culture treated on both sides for cell growth.
Endothelial cells are seeded onto collagen-coated inserts. The endothelial monolayer forms in several days, which occludes the membrane pores. The cell monolayer is then treated for 24h with Eyeseryl 1, 0.1 and 0.01mg/mL, or with Interleukin-1ß (100ng/mL), which due to its permeabilising properties is used as a positive control. Non treated monolayers are used as controls. After treatment, FITC-Dextran is added on top of the cells, allowing it to permeate through the cell monolayer. The extent of permeability can be determined by measuring the fluorescence of the plate well solution.
Results
Results prove that Eyeseryl is able to inhibit vascular permeability in a dose-dependent manner, reaching a 50% inhibition respect to controls with 1mg/mL (0.1%) Eyeseryl. This activity in vitro would be translated in vivo in a decrease of water accumulation in eyebags, since Eyeseryl avoids water leakage from blood vessels.
The graph above shows the inhibition percentage of the vascular permeability 10 minutes after adding FITC-Dextran on the endothelial cell monolayers treated with Eyeseryl (1, 0.1 and 0.01mg/mL).
Eyeseryl has been tested in vivo on a group of 20 female volunteers, aged 18 to 65. A cream containing 0.01% Eyeseryl was applied twice a day during 60 days.
Pictures were taken at 0, 15, 30, 45 and 60 days.
Several parameters were measured instrumentally or visually and physically evaluated by a dermatologist.
A dermatologist assigned a score to the decrease in eyebag puffiness by compared to the picture taken at time 0:
1 – no reduction, 2 – slight reduction, 3 – fairly good reduction, 4 – good reduction
The scoring table of the dermatologist is enclosed, as well as the statistical analysis performed on the data. Since the measurements are non parametric (which means we don’t know the underlying distribution of the variable measured), they are analysed using Friedman’s Test.
Puffiness under the eyes is greatly reduced, even after only 15 days: 70% of the volunteers had improved at Day 15.
Applying Friedman’s test, the data for every timepoint is determined to be significant, according to the following table:
Conclusions
Puffiness under the eyes is greatly reduced, even after only 15 days! - 70% of the volunteers had improved at Day 15.
At the end of the test, 95% of the volunteers had improved:
o 30% showed a slight improvement (grade 2) o 30% showed a fairly good improvement (grade 3) o 35% showed a good improvement (grade 4)
The elasticity measurements are performed with a Cutometer®. In elastometric measurement, the skin surface is aspirated from the depression induced by the machine into the aperture of the elastometer’s measuring probe. The depth of the skin penetration inside the probe is measured by an optic sensor. Cutaneous elasticity reflects the skin’s potential capacity (measured in mm) for retraction.
Cutaneous elasticity is defined as the ratio:
Elasticity = Uf-Ua/Uf
Uf = skin extensibility Ua = residual deformation
Elasticity represents the recovery degree of the maximum deformation reached, whose values range between 0 and 1 (maximum elasticity). The experimental measurements, as well as the statistical analysis performed on the data, are shown below.
The moisture measurements are performed with a corneometer (SKINLAB®). Corneometry measures how skin moisturization changes. The higher the water content of skin, the lower the skin dryness. Corneometry is an indirect method to measure skin moisturization. It is based on the electric properties of skin surface. Indeed both electric capacity and the conductance of biological tissue change according to the water content, i.e. they increase if the water content increases. The experimental measurements, as well as the statistical analysis performed on the data, are shown below.
IN VIVO – Cream containing 1% Eyeseryl Solution
Test was performed on 17 people, aged 34 to 54, using a cream containing 0.001% Eyeseryl, applied twice daily during 28 days.
Eyebag volume was measured using a technique called Fringe Projection where 3D images of the study areas are obtained with a FaceScanner and processed with the software Optocat (Breuckman, Germany – EoTech, France).
After 14 days, 63% volunteers had reduced their eyebags After 28 days, 70% volunteers had reduced their eyebags Eyebags were significantly reduced by almost 3% in only 15 days!
High resolution photographs of the dark circles were taken under polarised light and chromametry studies were performed
The ITA° (Individual Topographical Angle) categorises skin colour : Brown< 10° <tanned< 28°<normal< 41°<light< 55° <very light
ITA significantly increased (+3%) showing a slight lightening effect (at T14 and T28).
L* is luminance which represents relative brightness from total darkness (L*=0) to absolute white (L*=100), while a* is red/green color axis and b* is yellow-blue color axis.
L* was significantly increased at T14 (+1%)
Conclusion
Both results (eyebag volume and dark circles) are moderate but significant. These tests were done at a very low concentration (1% Eyeseryl) in order to define a minimum concentration for which an effect is measured.
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