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Hydromanil to improve skin moisture and anti-aging effect - clinical informationHydromanil is used with great success in many of our anti-aging skin care products, since it has proven to result in a far more moisturized skin – and this fact has been clinically proven while the molecule was developed.
Although this page is highly technical, we have included it on our website for the benefit of those individuals who wish to ascertain the value of our cosmetic ingredients, and the efficiency with which they work.
INTRODUCTIONHydromanil is composed of galactomannan hydrating molecules arranged in a three-dimensional matrix that retains galacto-manno-oligosaccharides. Galactomannans
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This novel structure is composed of large galactomannan molecules and small oligosaccharide molecules of different sizes according to their polymerization degree (p.d.): smaller (p.d=2-5), larger (p.d.=5-10).
Qualitative analysis of size distribution was carried out by means of Thin Layer Chromatography (TLC) using a monosaccharide (glucose), a disaccharide (lactose) and a purified known galacto-manno-oligosaccharide (61-a-galactosyl-ß-1,4-mannotriose -GM3) as standards.
The quantitative composition was evaluated by HPLC. The following table shows the results.
Hydromanil contains a total of 5-7.5% oligosaccharides. The most representative ones are the trisaccharide GM2, the tetrasaccharide GM3 and the disaccharide M2.
These oligosaccharides have also been qualitatively identified. We detected the presence of disaccharides and trisaccharides, with and without galactose constituents, derived from the specific hydrolysis of the galactomannan chains.
The most important ones are:
The qualitative analysis also reveals the presence of an heptamer, 63,64-di-a-galactosylß-1,4-mannopentaose corresponding to G2M5 although the proportion of this compound is lower (<0.5%).
Hydromanil acts at two different levels to achieve a combined double effect on the stratum corneum:
Since the galactomannans in the colloidal matrix cannot penetrate the skin because of their large molecular size, they remain on the stratum corneum surface exerting an immediate filmogenic moisturizing effect that gives the skin a more homogeneous appearance.
When Hydromanil is applied onto the skin, the polysaccharides in the matrix immediately restrain the evaporation of transepidermal water (kinetic factor) and additionally retain part of the water contained in the cosmetic product.
The presence of numerous free hydroxyl groups (OH) in the polyol chemical structure of galactomannan and galacto-manno-oligosaccharides results in the formation of numerous hydrogen bonds with the water molecules. It is a simple but efficient mechanism that retains the water already present and the water contained in the product.
The film that these compounds form onto the skin also contributes to reinforce the moisturizing effect because it reduces the loss of water through perspiration.
As the galacto-manno-oligosaccharides retained in the matrix are released, they penetrate the stratum corneum – because of their small size – and get integrated into the epidermis where they exert their cumulative moisturizing effect.
Continued applications of Hydromanil maintain the three-dimensional matrix on the skin for a longer time thus enabling the retained molecules to be sequentially released depending on their size.
The colloidal matrix creates a state of moisture continuity with the stratum corneum. A gradient in the concentration of oligosaccharides is then generated from the exterior to the interior of the stratum corneum, which starts the mobilization of oligosaccharides in favor of the gradient:
Thus Hydromanil is a sequential delivery system of hydrating agents into the stratum corneum, which results in a longer lasting moisturizing effect.
These small moisturizing agents integrated into the stratum corneum act synergically with the NMF to improve the amount of retained water (physiological factor). By moisturizing from the most interior layers, the moisturizing effect can be maintained longer.
In studies carried out by Bowstra et al. on the distribution of water in the stratum corneum under different levels of relative humidity, it was observed that water tended to be accumulated within the corneocytes located in the central layers, with same distribution that NMF.
This presence of water in the central layers is responsible for the flexibility and certain biochemical activity (desquamation) in the stratum corneum. Low moisturizing levels prevent the correct activity of the hydrolytic enzymes that coordinate the desquamation of the external layers of the stratum corneum. These layers keep together although not adequately cohesive, which gives rise to an abnormal thickening of the horny area and a sensation of roughness.
On the basis of its cumulative moisturizing effects during the days of application, Hydromanil also influences this process by restoring the necessary moisture levels to correct the bad enzymatic activity in the stratum corneum.
It is a regulatory action of the cutaneous desquamative process that eliminates the layers of swollen cells and leaves the stratum corneum cohesive and flexible thus improving the appearance and feel of the skin.
In order to evaluate the moisturizing efficacy of Hydromanil on the target area – the superficial stratum corneum – an in-vivo study was carried out by applying a 10% aqueous gel formulation on the forearms of 12 volunteers.
In order to assess the moisture level of the skin, we took instrumental measurements (Corneometer® y D-Squame®) and carried out a sensory evaluation by means of a questionnaire that the volunteers fulfilled by the end of the study.
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The moisturizing kinetics was studied by Corneometry, in order to measure the moisture level at time 0 (T0), 30 min, 2 h and 4 h after a single application of the product formulated 2%, 5% and 10% in aqueous gel.
On application of Hydromanil formulated with the highest dose (10%), the concentration of active compounds was high enough to retain temporarily a part of the water contained into the cosmetic formulation. A maximum 30.8% increase in moisturizing was recorded after 30 min.
Later readings (2h, 4h) revealed that transepidermal water accumulated without dissipation. However, the water contained in the product had been partially lost. The result was a moderate decrease to 21% after 4 hours.
With medium and low concentrations (5% and 2%) the instantaneous effect at 30 min is not so marked, which reinforces the importance of the transepidermal water retention that results from the filmogenic effect of Hydromanil. Under these conditions, observations at later times revealed an increase in the moisturizing level due to the accumulation of transepidermal water without dissipation (kinetic factor).
In order to assess the long lasting moisturizing effects, the study was extended to 21 days. The volunteers applied the formulations (Active 10% and Placebo) twice a day. The figure below shows the averaged results recorded on the day following to the end of the treatment (Day 22).
The moisturizing level of the Hydromanil treated area on day 22 was clearly higher (+12.5%) as compared with the initial values of each volunteer before the start of the study (D0/T0). These results demonstrate that repeated applications of this product have cumulative moisturizing effects as compared with the placebo.
The desquamation index was calculated by means of the D-Squame®. The figure below shows the results.
Hydromanil produced an important and statistically significant 60% reduction of desquamation. These results demonstrate its ability to regulate the desquamation process and to restore cellular cohesion in the outer layers of the stratum corneum.
The figure below shows that all of the volunteers treated with Hydromanil experienced a reduction in desquamation. On the contrary, the Placebo treated area showed a general worsening of desquamation.
Besides the instrumentally recorded results, the volunteers carried out a sensory evaluation of the efficacy of Hydromanil.
Remarkably, 83% of the volunteers reported smoother and finer skin on the Hydromanil treated area and 67% of them reported better moisturized skin.
Their evaluations agreed with the desquamation index results, which evidenced the change from dry rough skin with a thick poorly cohesive stratum corneum to smoother better-moisturized skin with a cohesive stratum corneum and normal desquamation.
Hydromanil is a novel moisturizing active compound that, because of its innovative structure and production technology, acts as a Sequential Delivery System of small molecules, resulting in a final combined moisturizing effect: immediate moisturizing and long-lasting moisturizing.
Its immediate moisturizing effects make it suitable for basic moisturizing formulations aimed at body or facial care such as shower gels, body milks and hand creams.
Its cumulative and long-lasting moisturizing effects make it suitable for specific products to treat dry, damaged or aged skin.
Its desquamation regulatory activity and cellular cohesion restoring properties make it suitable for formulations aimed at improving the skin appearance (smoother, finer, more flexible).
Its high moisturizing capacity also makes it suitable for hair products, like shampoo or hair lotions, to improve some physical properties of hair, such as fragility.
High Performance Liquid Chromatography (HPLC) using a Beckman System Gold chromatograph equipped with a refractive index detector (Beckman, Model 166). The mobile phase was acetonitrile:water (60:40, v/v); the stationary phase was an Spherisorb5-NH2 column (250 x 4,5 mm ø). The flux was 1.0 mL/min and the sample injection volume was 20µL. We used standards of every sugar to be analyzed. Calibration curves were constructed with different water solutions of these sugars within a 0-5 mg/mL range.
Official method nº 920.52 of the AOAC (Association of Analytical Chemists). Briefly, 25g of sample were hydrolyzed with 15mL HCl (37%) for 2 hours in reflux boiling and subsequently refrigerated and neutralized with a 10% NaOH solution. Inversion and quantification of sugars was then carried out according to the Luff-Schoorl method, using a 25 mL sample aliquot. The results were expressed as % glucose.
Sample preparation: 4g sample were dried and resuspended in 3 ml water. The mobile phase was a mixture of butanol:ethanol:water (50:50:30), the stationary phase was a silica gel 20cm x 20 cm plate. Analyzed sample: 10 µ. Detection: diphenylamine reagent (2.4g diphenylamine + 2.4aniline-HCL were dissolved in 200mL methanol and 20mL 86% phosphoric acid were added). 10mL reagent were sprayed on the plate, heated to 105ºC for 5min and observed under visible light.
Cytotoxicity was evaluated on cells of the line SIRC and the concentration causing 50% mortality (CI50) was determined by means of the Neutral Red Release assay. On the basis of the average of two CI50 determinations, the product can be classified according to its cytotoxicity (Journal Officiel de la République Française, December 30th, 1999). The CI50 was >50%. The results recorded under the present experimental conditions indicate that Hydromanil did not exert cytotoxic effects.
This test evaluated the compatibility of the product with the skin after a single 48 hours application on the skin of the back of 10 adult volunteers. The irritant effect was clinically evaluated 30 min after removing the patches. A 0.50 irritation index was recorded. The results obtained under the present experimental conditions indicate that Hydromanil is well tolerated by the skin.
This assay evaluated the absence of potential skin irritant agents by repeated applications of occlusive patches on the skin of 50 healthy adult volunteers of both sexes. Clinical observations were carried out after removing the patches during the induction phase and the challenge phase. Neither pathological irritation, nor sensitization reaction significant of a cutaneous intolerance was noted. From the results obtained under the experimental conditions, Hydromanil can be considered non sensitizing.
Visual observation of the effect of a single five minutes application on the chorioallantoic membrane according to the Journal Officiel de la République Française, December 26th, 1996. The irritation index of Hydromanil was 3.0, which classifies this product as a mild irritant agent.
The potential ability of this product to induce reverse mutations in Salmonella typhimurium with and without a metabolic activation system (S9 mix) was assessed. The salmonella strains TA-1535, TA-1537, TA-98, TA-100 and TA-102 were treated with six different doses of the product (Directives OECD, nº 471 and CEE B13/B14, June 8th, 2000). No mutagenic response was observed for any of the bacterial strains, at the concentrations tested, with or without the addition of S-9. Under the experimental conditions, Hydromanil did not show mutagenic activity in the bacterial reverse mutation test.
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