The present invention relates to a preparation for dermatological treatment, particularly for skin diseases and for cosmetic skin treatment and to a process for producing said preparation.

The aim of the present invention is to provide a functional ingredient of great cosmetic interest, with a sure efficacy and that allows natural raw materials to be used that compose at least partially biomasses currently not used for such purposes.

With reference to the above mentioned objects, therefore the invention relates to a preparation for dermatological treatment, particularly for skin diseases and/or for cosmetic skin treatment comprising active ingredients from extract of vegetable material, wherein said vegetable material is composed of the plant called as Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean Sea.

Particularly said preparation comprises an extract from vegetable material, wherein said vegetable material is composed of biomass of Posidonia on beaches, particularly Posidonia oceanica, endemic to the Mediterranean Sea.

According to a further aspect of the present invention, the use of said preparation is not limited to the cosmetic field, but it can be used also for producing other types of preparations such as for example pharmaceutical, nutraceutical preparations, in dietary supplements, infusions, and in functional food.

The invention is based on the fact of recognizing that Posidonia seagrass not only has high concentrations of iodine and other compounds common to algae, but unlike the latter it further has a high contents of cichoric acid and other active ingredients that exhibit biological properties functional for antioxidant, anti-aging, depigmenting, anti-cellulite actions and for the treatment of other skin disorders.

Therefore it is a unique starting material, that combines the fact of being rich in elements such as iodine, typical of marine flora, also with the fact of being rich in secondary metabolites, such as phenolic compounds, among which, particularly, chicoric acid.

This is made possible by the fact that Posidonia is a superior plant living in a marine environment, similar to the one where many algae live (Fucus, Laminaria, etc) , and that therefore is characterized by high concentrations of iodine. At the same time Posidonia is able to synthesize chicoric acid, by means of biosynthetic pathways belonging only to superior plants (e.g. several Ferns, sea phanerogams, Lamiaceae and above all Asteraceae) .

Posidonia oceanica that lives in the protected marine area of Egadi Islands is particularly rich in chicoric acid.

The use of plants deriving from such area has the great advantage that water clearness and the absence of pollutants, on one hand allow the plant to perform the typical cycles of superior plants and to perform an effective synthesis of substances important for cosmetic purposes and on the other hand prevent pollutants or allergenic substances such as Nickel or other metals from being present.

Posldonla oceanlca (L.) Delile is a sea phanerogam endemic to the Mediterranean Sea. Such plant has rhizomes with tufts of 6-7 ribbon-like leaves, that can reach a length greater than one meter. The species forms large sea meadows along the coasts, from the surface up to a depth of about 40 m, and it has an important role in protecting them from marine erosion, in addition to be a key element for preserving Mediterranean ecosystems.

After sea storms, accumulations of Posidonia on beaches form banquettes, up to 2 m in height, at water- beach interface and in the first meters of emerged beach. The accumulations can also be distributed on large areas of coastline, in the order of many hundreds of meters and up to 15 towards inland. Such material has to be handled with precise rules such to guarantee the presence of enough banquettes such to oppose phenomena of coastline erosion.

At the same time, an amount of the material on beaches usually is removed in order to allow the beach to be used for entertainment purposes and touristic activities and for usual harbor and fishing activities.

Such amount of residues on beaches , up to recently, has been used as waste, or at most it has been used as amendment for composts (D.M. 217/2009) . By means of the present invention such material, that is the biomass of Posidonia on beaches that is collected such to clean beaches, is converted from waste into a resource that can be subjected to eco-compatible exploitation resulting in not negligible economic implications, when the precious metabolites present therein are suitably extracted and re-used.

Particularly the present invention provides to use Posidonia extract as specified above not only for cosmetic purposes but also as an active ingredient for dermatological treatment.

Posidonia extract, by means of its high contents of chicoric acid, is particularly efficacious in promoting biochemical processes for repairing skin cell damages. Skin is composed of keratinocytes , fibroblasts, melanocytes, immune T-cells etc. and it is known that many factors, such as strong UV radiation, pollutants dispersed in the environment and physical stress, can result not only in natural aging process but also in damages to DNA of such cells and can affect their circadian rhythm. According to what mentioned by some authors and confirmed in documents US2014/0242010A1 and US2015/0071895A1 , chicoric acid is an effective stimulator of perl gene expression in skin cells and this should led to a better synchronicity in the response thereof. When such cells exhibit same level of perl gene expression, treatment of the skin with an active ingredient able to repair cellular damage is more effective.

Previous chemical analyses carried out on Posidonia leaves pointed out the presence of high amounts of iodine and polyphenol derivatives of benzoic and cinnamic acid, among which chicoric acid, caffeic and ferulic acid; moreover flavonoids, steroids, essential fatty acids and sesquiterpenes have been found.

Therefore Posidonia extract is a complex cosmetic ingredient, that is a mixture of organic and inorganic compounds and substances that synergically contribute in exerting an overall cosmetic effect with several functionalities .

Among the listed elements and active ingredients, in cosmetic applications iodine is generally used as anti-cellulite ; ferulic acid (INCI name) is generally used as UV filter, antioxidant, with depigmenting effect, with anti-aging effect, antimicrobial and lenitive effect; caffeic acid (INCI name) , chicoric acid being a precursor thereof, as antioxidant, antiinflammatory, for UV protection, antimicrobial and depigmenting effect; chicoric acid as antioxidant, with depigmenting effect and as immunostimulant.

Therefore the cosmetic treatment can be performed by using the extract, that is by applying on the skin said extract as ingredient of a cosmetic preparation in the form of cream, emulsion, milk, gel, slurry, lotion.

In these cases substances typically used for the formulation of such type of preparations are added to the ingredient composed of Posidonia oceanica extract. The composition of Posidonia extract makes such extract also functional to the use in preparing drugs and nutraceutical products, such as also dietary supplements, infusions, and functional food.

Also in this case the use of the extract is provided in combination with excipients and other substances usually used in such products.

The document GOKCE ET AL: "Evaluation of antidiabetic antioxidant and vasoprotective effects of Posidonia oceanica extract" , JOURNAL OF

ETHNOPHARMACOLOGY. ELSEVIER IRELAND LTD. IE. vol 1. 115. no. 1, 28 November 2007 (2007-11-28) , ISSN : 0378- 8741 evaluates the power of Posidonia extract in restoring levels of typical cell antioxidants, such as glutathione, superoxide dismutase, catalase and glutathione peroxidase in diabetic rats. Therefore the ability of restoring body antioxidant defenses affected by oxidative stress is measured.

Biological tests described in the document have been made on alloxan diabetic rats, by measuring the ability of orally administered Posidonia extract to limit the level of plasma glucose. Moreover analyses of liver and kidney functionality have been performed in the same animals. In vitro aorta preparations obtained from the same animals have been used to evaluate the effects of the extract on vascular contraction. No studies have been made as regards cytotoxicity of the hydroalcoholic extract of Posidonia oceanica leaves, and no studies have been made for evaluating the depigmenting ability of the extract by tyrosinase inhibition and reduction of melanine in melanocytes and no evaluations have been made about the ability of the extract in promoting the growth and production of collagen as well as no evaluations have been made about the ability of the extract in stimulating the lipolysis .

In order to obtain the cosmetic ingredient and to carry out the extraction process it is possible to follow different extraction methods known in prior art.

The following example is one of the possible methods for extracting the cosmetic ingredient from Posldonla oceanlca .

A predetermined amount of fresh Posidonia oceanica, just collected from seashore, is deprived from the base sheath and cleaned from epiphytes present on the leaf blade. Leaves treated in this manner are dried for example in a ventilated oven and protected from light.

At the end of drying process, the dried leaves are ground by means of an industrial grinder to obtain a mass of finely ground material.

The grinding step is controlled by adjusting the temperature, such to keep it within a predetermined upper limit. Moreover it is necessary to reduce as much as possible the time vegetable material is exposed to light, to avoid a decrease in yield of noble products.

The ground material therefore is subjected to extraction by solvent.

It is possible to use water solutions of different solvents such as for example a ketonic solvent and carbossylic acid, for example acetone and formic acid or preferably a hydroalcoholic solvent.

Extraction step lasts several hours and it is performed under stirring and at room temperature.

Mixture is then filtered to reach a clear solution; it is then dried under reduced pressure, at a predetermined temperature.

The resulting solid can be washed with acidulated water in suitable proportions with respect to the dried substance and finally it is again vacuum dried.

From analyses made by high-performance liquid chromatography together with mass spectrometry (HPLC- MS) the more abundant signal is the one of chicoric acid, both in extracts with acetone and water and in those obtained by using methanol and water or ethanol and water. The presence of chicoric acid was ascertained by comparing the chromatography retention time and the fragmentation spectrum with those of a separately injected standard.

There is also a signal, less deep than the one of chicoric acid, that is referable to caffeic acid or to a precursor thereof.

According to a preferred embodiment the extract mainly comprises the presence of chicoric acid and caffeic acid in a chicoric acid/caffeic acid ratio of 20/1.

According to one embodiment the extraction provides to use water and ethanol in 40/60 ratio, and brought to pH=4 with formic acid. Dried extract is obtained by evaporation of solvents . Analyses show a very high production of chicoric acid, in the extent within the range 10-100 mg/lOOg by weight of dry extract.

50 grams of Posidonia oceanica leads to a volume of filtered final extract of about 900ml and to a concentration of chicoric acid of 0.55mg/ml.

Iodine content is in the order of some dozens of ppm, that is from about 40 to 80 ppm.

Analyses about the power of Posidonia extract showed an antioxidant action in the order to 500 to 600 (mM) expressed in gallic acid equivalents.

According to a further aspect the invention relates to an antioxidant preparation based on ethanol extract of the plant called as Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean sea.

Said extract particularly as an antioxidant action of about 500 to 600 (mM) expressed in gallic acid equivalents .

A further experimentally ascertained power of extract of Posidonia Oceanica seagrass is depigmenting activity.

A further aspect of the present invention therefore is about a preparation having depigmenting effect by tyrosinase inhibition based on extract of the plant called as Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean Sea.

Said extract has particularly a dose-dependent inhibition action of mushroom tyrosinase, with 20% inhibition for a dose of 5 micrograms/ml and about a 45% inhibition with 1000 micrograms/ml .

A further experimentally ascertained action is melanogenesis inhibition.

A further aspect of the present invention therefore is about a preparation with functions for treating disorders in skin pigmentation, based on extract of a plant called as Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean Sea.

Typical disorders of pigmentation that can be treated by the preparation of the present invention for example are disorders such as melasma and senescence staining, freckled skins, postinflammatory hyperpigmentation and actinic damage sites.

The study about the functionalities of the preparation based on extract of Posidonia oceanica plant further showed an activity stimulating fibroblasts and collagen production.

Fibroblasts are the main cells of the derma and responsible for the production of dermal matrix. Such cells are essential for keeping the skin tone, elasticity and firmness, and to prevent defects such as fine lines, wrinkles, skin sags and stretch marks.

Collagen is the main component of dermal matrix, where type 1 collagen forming bundles of fibers is the more abundant isoform. Collagen is produced by fibroblasts and it is fundamental for skin tone and firmness. Under normal conditions, collagen is subjected to a renewal process consisting in the continuous degradation by matrix metalloproteases and the production by fibroblasts. During aging, degradation processes of collagen tend to dominate the renewal, causing wrinkles and other skin alterations. Therefore the fact of maintaining the function of fibroblasts is a pre-requirement for delaying skin aging .

A further aspect of the present invention is therefore about a preparation with functions for stimulating fibroblasts and increasing collagen production, based on extract of the plant called Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean Sea.

The increase of collagen production is higher than 10% with doses of 10 and 20 micrograms/ml .

Still another action exerted by the preparation of the present invention is about the lipolytic action.

Dose-dependent lipolytic effect of the hydroalcoholic extract of Posidonia oceanica leaves suggests its possible use in anti-cellulite cosmetic products .

Therefore still another aspect of the invention is to provide a preparation with functions for stimulating adipocytes and increasing adipolysis induction for anti-cellulite treatment, based on extract of the plant called Posidonia, especially Posidonia oceanica, endemic, that is typical, to the Mediterranean Sea.

The invention will be shown below with reference to some experiments whose results are shown in the annexed figures wherein: Fig.l is a graph about verification of antioxidant action of the preparation according to the present invention .

Fig.2 is a graph about tyrosinase inhibition action of the preparation according to the present invention .

Fig.3 is a graph about verification of melanogenesis inhibition action of the preparation according to the present invention, where arbutin is used as positive control.

Fig.4 is a graph about stimulation of fibroblast growth rates obtained with different doses of the preparation according to the present invention.

Fig.5 is a graph about the collagen production obtained by the treatment with different doses of the preparation according to the present invention.

Fig.6a is a graph about glycerol standards used to quantify the action of the extract on lipolysis.

Fig.6b is the effect of increasing doses of the preparation according to the present invention on adipocytes in terms of lipolysis measured as the release of glycerol with respect to the control.

An extract of Mediterranean Posidonia Oceanica endemic to the Mediterranean Sea was used in tests below.

The extract was obtained by a process in hydroalcoholic solution.

The process provided the following steps:

plants were dried for about 36h at a temperature of 42°C. once drying ended the material was ground such to obtain a pulverized material with grain size of 1-2 mm; a mixture with 60% of bio-ethanol, 39.7% of H2O and 0.3% of H-COOH was prepared by means of which extraction was carried out;

extraction was carried out by using 1 liter of solution for lOOg of dry material at room temperature or under a very slight heating (30 °C) for 120 minutes under stirring and while maintaining pH=4 by using H- COOH.

At the end of 120 minutes the supernatant was removed and a second extraction was carried out as described above.

Supernatants obtained from the two extractions were gathered together, subjected to rough filtering to recover the solvent and to proceed with vacuum filtration with sintered glass filter. The organic solvent was taken to dryness under vacuum maintaining the temperature lower than 45° in a rotary evaporator.

The precipitate was recovered, weighed and dried under vacuum (lyophilizer) for removing solvent residues and then it was finely pulverized obtaining the dry preparation in powdery form of the precipitate.

By using the hydroalcoholic extraction process described above it was noted that pH remained constant during the treatment without the need of being regulated by formic acid. It is probable that the presence of ions and acids in the biological material allowed a buffer solution to be obtained.

The extract obtained in this manner was subjected to biological tests to verify pharmaceutical and cosmetic properties particularly for dermatologic and cosmetic treatment of the skin.

Test 1 Antioxidant power

Reactive oxygen species (ROS) are extremely reactive compounds generated from metabolism of oxygen in human beings, due to chain reactions of free radicals. Such chemical agents, when produced in excess, are considered as performing an important role in tissue degenerative processes, for example in pathologic conditions and during ageing. On the basis of such assumptions interest has been focused on compounds present in the diet or anyway biocompatible compounds, preferably of natural origin, able to inactivate free radicals and to act as antioxidants.

The antioxidant power of hydroalcoholic extract of Posidonia oceanica leaves was evaluated by the usually used DPPH (2 ,2-diphenyl-l-picrylhydrazyl) assay.

An hydroalcoholic extract of goji barriers (Lycium barbarum) , a natural product with a high antioxidant power, was used as positive control.

The test was performed as it follows:

Extracts were diluted in 1:10 in ultrapure water, mixed 1:1 with 60 μΜ DPPH in absolute ethanol , incubated at RT for 15 min in the dark, and then absorbance of reaction mixture was measured at 520 nm in a microplate reader VMax (Molecular Devices, Sunnyvale, CA) . Absorbances of serial dilutions of Gallic acid were used to obtain a standard calibration curve, and antioxidant power of extracts was therefore expressed as Gallic acid equivalents (mM) . Data are shown in the graph of figure 1 and show that Posidonia extract has antioxidant power comparable to that of Goji berries, showing a high antioxidant power of Posidonia .

Toxicity tests by cell viability assay of cell cultures in vitro

Cell cultures in vitro were made composed of stabilized human cell lines of keratinocytes (HaCaT) and skin fibroblasts (46BR.1N). Cells were cultured in vitro in DMEM, supplemented with 10% fetal bovine serum (FBS, Sigma), 1% glutamine, and 1% penicillin- streptomycin solution (Sigma) at 37 °C in humid atmosphere containing 5% C02.

Cell viability assay was performed as it follows:

The first step for studying biological properties of a substance is evaluating its toxicity, in order to select subtoxic doses to be used in following analyses.

Cytotoxicity of hydroalcoholic extract of Posidonia oceanica leaves was determined in keratinocytes and fibroblasts using the standard MTT assay, based on the reduction of tetrazolium reactive MTT, (3- ( ,5-Dimethylthiazol-2-yl) -2,5-

Diphenyltetrazolium Bromide, in insoluble formazan operated by mitochondrial enzymes. Cells were settled in 96-well plates for 24 h, 10,000 cells for well, and therefore exposed for 48 h to increasing logarithmic series of concentrations of the extract in DMEM, ranging between 1÷1000 g/mL. Then cells were incubated for 3 hours at 37 °C with a solution composed of 100 microliters MTT (5 mg/ml in PBS) per ml of cell culture medium without serum, and then treated with IN HC1- isopropanol solution (1:24, v/v) followed by mixing to dissolve the generated dark blue crystals of formazan. After few minutes at room temperature, plates were read at 550 nm in microplate reader.

Dose-response curves obtained from MTT data were analysed by a logistic regression model, allowing the following cytotoxicity parameters for the extract to be estimated: IC50 = 115 g/ml and IC05 = 25 g/ml on fibroblasts; IC50 = 160 g/ml and IC05 = 35 g/ml on keratinocytes . Where IC50 = dose causing 50% inhibition of cell viability; IC05 = dose causing 5% inhibition of cell viability. IC05 is usually taken as toxicity threshold in toxicological studies.

Test for depigmentation activity by measuring in vitro tyrosinase inhibition and melanogenesis inhibition

In vitro tyrosinase inhibition

The activity of tyrosinase enzyme is an important step in melanin synthesis in melanocytes. Depigmenting properties of hydroalcoholic extract of Posidonia oceanica leaves were evaluated using an in vitro test of mushroom tyrosinase inhibition (Kobayashi et al 1995. Biosci Biotechnol Biochem 59:. 1745-1746), a common screening of tyrosinase inhibitors.

Aliquots of 10 mL of a solution composed of 125 U/mL of mushroom tyrosinase (Sigma-Aldrich) in phosphate buffer (pH 6.8) were added to 96-well plates, followed by 70 ml of phosphate buffer and 60 ml of ultrapure water or extract dissolved in ultrapure water, to obtain a series of final concentrations ranging between 5÷1000 g/ml of extract. For the positive control kojic acid was used instead of the extract. Then 70 L of L-tyrosine (Sigma-Aldrich) were added at a concentration of 0.3 g/ml in ultrapure water. Blanks without enzyme were included for all the conditions. Plates were incubated at 30°C for 30 min and the absorbance was read at 505 nm in microplate reader. The inhibitory activity (IA%) percentage was calculated according to the formula:

IA%=(1- (Abs (ex+en) -ABS (ex) ) / (Abs (en) -ABS (bk) ) ) *100 where :

Abs (ex+en) = absorbance of mixture with extract and enzyme ;

Abs (ex) = absorbance of mixture with extract and without enzyme

Abs (en) = absorbance of mixture with enzyme and without extract;

Abs(BK)= absorbance of mixture without enzyme and without extract (blank) .

The kojic acid positive control exhibited a dose- dependent inhibition of tyrosinase activity with IC50 of 4.7 g/ml . Data about hydroalcholic extract of Posidonia oceanica leaves, summarized in figure 2, show a dose-dependent inhibition, from about 20% inhibition at 5 g/ml up to about 45% inhibition at 1000 g/ml . For all the groups, test t with Bonferroni's correction provided values of p<0.01 (n = 6) in the comparison with the control (null inhibition) . Melanogenesis inhibition

Cells of human melanoma cell line MEWO were cultured in RPMI medium supplemented with 10% (v/v) FBS, 1% glutamine and 1% of penicillin-streptomycin solution (Sigma) , at 37 °C at humid atmosphere with 5% C02. Cells at confluence were resuspended and plated in 96-well plates to evaluate the extract cytotoxicity as described above.

MTT assay performed after 72h exposure allowed IC50>500 g/ml and IC05XL00 g/ml to be evaluated. Then cells were settled in 24-well plates (100,000 eelIs/well) for 24 h, and exposed to the extract at the dose of 50 g/ml for 72 h.

A series of samples treated with Arbutin (1 mg/ml) as positive control were used. After treatment, the contents of melanine of the cells was determined by removing the culture medium, by washing the cells with PBS, by trypsinization , centrifugation , by removing the supernatant and collecting the cell pellet subjected to freeze-thaw cycle.

Therefore the cell pellet was dissolved in 100 L of 1 N NaOH and the raw cell extract was analysed with the microplate reader at 600 nm to determine melanine contents .

Results of figure 3 are expressed as percentage amount of melanine with respect to control cells (* = p <0.01, t test) . All tests were performed in triplicate.

These results suggest a possible use of the extract in depigmenting formulations for the skin.

The above tests made it possible to show how as a whole the depigmenting activities of the plant suggest a possible use of the extract in formulations for treating skin pigmentation disorders such as melasma and senescence spotting, freckled skin, postinflammatory hyperpigmentation and actinic damage sites .

Test of fibroblast stimulation and collagen production

Fibroblast proliferation

Fibroblasts are the main cells of derma and are responsible in the production of dermal matrix. These cells are fundamental for maintaining tone, elasticity and firmness of the skin, and to prevent defects such as fine lines, wrinkles, skin sagging and stretch marks .

Effects of the hydroalcoholic extract of Posidonia oceanica leaves on fibroblast growth rate were evaluated by settling cells in 96-well plates as above, except that 5,000 cells per well were used. Fibroblasts were treated for a period of 10 days with 10 or 20 g/ml of extract. At the end of exposure, MTT assay was used to evaluate the density of viable cells, obtaining cellular growth curves. Data showed a dose-dependent increase of cellular growth of fibroblasts exposed to the extract. Figure 4 shows curves of growth rates of fibroblasts expressed as mean ± S.D. of cell density in arbitrary units (n = 8) as a function of time (* = p <0.01, t test) .

Collagen production

Collagen is the main component of dermal matrix, where type 1 collagen forming bundles of fibers is the most abundant isoform. Collagen is produced by fibroblasts and it is fundamental for skin tone and firmness .

Under normal conditions, collagen is subjected to a renewal process that is the continuous degradation by matrix metalloproteases and the production by fibroblasts. During aging, degradation processes of collagen tend to dominate the renewal, causing wrinkles and other skin alterations. Therefore the fact of maintaining the function of fibroblasts is a pre- requirement for delaying skin aging.

The effect of the extract on the production of type I collagen by fibroblasts was evaluated by using ELISA technique.

Fibroblasts were settled in 96-well plates (15,000 eelIs/well) for 24 h, and exposed to 5 , 10 or 20 g/ml of extract for 72 h. After the treatment, the medium was removed and cells were washed with PBS (100 L/well) , fixed with 3.7% paraformaldehyde for 10 minutes, washed 3 times with washing buffer (0.5 mM CaC12, 1 mM MgC12 , 0,1% Triton in PBS) (100 L/well) , incubated with blocking buffer (3% BSA in washing buffer) for 30 minutes, washed with washing buffer, incubated with human anti-collagen mouse primary antibody (ab6308, Abeam) diluted 1:300 in washing buffer containing 1% BSA (50 L/well) for 2 h under stirring, washed 3 times with washing buffer, incubated with rabbit anti-mouse IgG HRP-conjugated secondary antibody (ab97046, Abacm) diluted 1:1000 in washing buffer containing 1% BSA (50 pL/well) for 60 minutes under stirring, and washed 3 times with washing buffer.

Then the solution was accurately removed from the wells, plates were incubated with IStep™ Ultra TMB ELISA substrate solution (Thermo Scientific Pierce, 50 ml / well) for 15 minutes, blocked with 2 M sulphuric acid, and read at 420 nm in microplate reader.

All the procedure was performed at RT.

Data of figure 5 show a considerable increase higher than 10% of collagen production in fibroblasts exposed to 10 and 20 g/ml of extract, with respect to cells maintained under control conditions (* = p <0,01, t test) .

The whole effects on fibroblast growth rate and collagen synthesis denote that hydroalcoholic extract of Posidonia oceanica leaves exerts a deep stimulating effect on fibroblast activity, suggesting it to be used in anti-wrinkle and anti-aging formulations for skin treatment .

Lipolysis test

Cellulite is a skin condition associated with localized fat deposits, on the surface having the characteristic orange peel aspect. Adipocytes are cells storing fats of the adipose tissue that can be subjected to an excessive deposit of lipid material associated to the appearance of cellulite. Lipolytic agents topically applied can be able to limit fat depositing in adipocytes and to reduce cellulite flaws.

Lipolytic activity can be evaluated in vitro by measuring glycerol released in the medium by adipocytes after triglyceride degradation, the main lipidic component of cellular deposits.

Therefore the lipolytic effect of the hydroalcoholic extract of Posidonia oceanica leaves was evaluated on human primary adipocytes by using ZenBio Cellulite Treatment Screening Kit Human Adipocyte Lipolysis Assay (Zen-Bio Inc., Research Triangle Park, NC, Cat # LIP-12) for finding free glycerol.

Following the supplier's protocol, cells were incubated for 3 h with the extract, and samples of conditioned culture medium were then analysed for the glycerol contents .

Data of figure 6a show a curve of glycerol standard while figure 6b show the effect of increasing doses of extract on adipocytes, expressed in terms of lipolysis fold induction calculated as ratio of mols/l of glycerol released by treated cells and by controls (* = p < 0.05) .

The dose-dependent lipolytic effect of hydroalcoholic extract of Posidonia oceanica leaves suggests its possible use in anti-cellulite cosmetic products .

Whole data obtained from the above disclosed tests are an absolute novelty in studies about Posidonia oceanica and set forth a series of unknown biological properties and that make the plant very interesting for developing pharmaceutical preparations for dermatological treatments and for skin diseases as well as for skin cosmetic applications, as antioxidant, anti-aging, depigmenting and anti-cellulite, suitable for treating skin and for treating skin disorders .