Sustainable cosmetic films

Abstract

A cosmetic solid composition comprising: a. phospholipid nanoparticles comprising a product obtained by extraction of a vegetal matrix with a hydroalcoholic mixture or with biosolvents; and b. a natural film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof; and optionally c. a plasticizing agent, preferably glycerin; wherein said phospholipid nanoparticles are preferably ethosomes, phytosomes, liposomes and wherein said composition can be reconstituted into a fluid cosmetic by the addition of water at the time of application on the skin, wherein said composition is anhydrous, wherein said composition does not comprise preservatives, wherein said composition is in the form of a film.

Description

[0001] SUSTAINABLE COSMETIC FILMS

[0002] FIELD OF THE INVENTION

[0003] The present invention falls within the skincare sector and concerns solid or waterless cosmetic products. The invention is particularly directed to sustainable cosmetic films (SCF) based on ingredients capable of being reconstituted and transformed into a gel or emulsion upon application. The SCFs of the invention can be functionalised with different types of extracts depending on the desired cosmetic actions (anti-aging, moisturising, soothing, anti-spotting, antiseborrhoeic, anticomedogenic, purifying actions).

[0004] TECHNICAL BACKGROUND

[0005] The formulations called "Sustainable Cosmetic Films" (SCF) arise from the need to meet the demand of the cosmetic / nutraceutical / pharmaceutical market that is increasingly oriented towards the search for an effective but also environmentally friendly product. The innovative aspects of SCFs concern the choice of raw materials and above all the way in which cosmetics are packaged / applied. Current trends in the cosmetic market identify some consumer preferences such as the preference for ingredients of natural origin, the high safety and tolerability of products linked to the reduction of excipients and synthetic preservatives, the environmental, social and economic sustainability of the entire production chain of a cosmetic.

[0006] In this regard, an exponentially growing trend is that of the so-called "solid cosmetics" or "waterless" welcomed above all by the new generations much more sensitive to environmental protection. Waterless products are indeed free of water and this entails an advantage for the user and for the environment. In fact, the absence of water does not allow microbial growth and for this reason it is possible to predict the reduction of the use of synthetic preservatives often responsible for the appearance of sensitizations and allergies. On the other hand, a waterless product is a much lighter product than a water-containing product, taking into account that on average the water content in cosmetics is often above 70%. This implies a considerable reduction of costs and environmental impact due to the transport of products. Another reduction in the environmental impact of waterless cosmetics derives from the possibility of using much more eco-sustainable packaging since being free of water they can be packaged in paper avoiding bottles and jars and avoiding the use of plastic.

[0007] International Patent Application No. WO2014 / 165490 discloses a liposomal composition comprising phospholipids and plant extract of Lupinus albus seeds, wherein said plant extract is obtained by a process employing ethanol, and said liposomal composition is combined with a composition comprising xanthan gum and glycerol, or glycerin to obtain as a final product a skin cream in solid form which when mixed with water takes the form of a fluid.

[0008] Chinese Patent Application No. CN111773118 discloses a powder cosmetic composition containing nano-liposomes comprising grape seed oil and lecithin, wherein the plant extract is obtained by a process involving organic solvents such as ethanol. Said composition also contains carrageenan and xanthan gum as well as octyldodecanol and pentane as plasticizing agents and is in the form of a powder which takes the form of a suspension when mixed with further water.

[0009] Chinese Patent Application No. CN108904369 describes liposomes comprising lecithin, sodium hyaluronate and Gynostemma pentaphyllum extract, which are mixed with glycerin and other excipients and then lyophilized to obtain powder. Said lyophilized powder is then mixed with water to obtain a fluid that can be applied as a cosmetic mask on the face.

[0010] Chinese Patent Application No. CN113425634 discloses a face mask comprising liposomes containing Ganoderma lucidum extract, the latter obtained by cold extraction in the presence of ethanol. This mask is reconstituted in a fluid with added water at the time of use.

[0011] Although there are waterless cleansing products on the market, such as soaps, shampoos, bubble baths, toothpastes, or oils and butters for the body, or face masks, to date waterless products for facial skincare, in particular gels and emulgels, do not seem to be equally available. Therefore, in this specific sector there remains a need for a waterless product of natural origin, characterized by a high safety and tolerability profile thanks to the low content of excipients and synthetic preservatives, with a lower weight than traditional products and with a low environmental impact.

[0012] SUMMARY OF THE INVENTION

[0013] The present invention provides a solution to the aforementioned technical problem by providing specific SCF formulations. The advantages that the SCF formulations object of the invention offer to the consumer lie, for example, in the possibility of having a product obtained from natural ingredients, without preservatives, preferably synthetic, and therefore highly tolerable, easily transportable and with a small footprint; the advantages for the manufacturer lie, for example, in the possibility of having an innovative and sustainable product, easy to scale, and a considerable reduction in packaging and transport costs; the advantages for the environment lie, for example, in the reduction of the use of synthetic ingredients, water and the carbon footprint linked to transport, and the possibility of using lightweight and biodegradable materials for packaging. SCFs represent the first opportunity to have waterless cosmetic gels and emulgels available. SCFs are obtained from ingredients of natural origin, free of water and the absence of water makes it possible not to use preservatives. SCFs are quickly rehydratable with the addition of a small amount of water before use. Their formulation has been designed from natural ingredients and analysed and characterised from a chemical-physical, technological and rheological point of view.

[0014] The potential of SCFs is also linked to their high versatility; in particular, they may contain different bioactive substances depending on the desired cosmetic action. In fact, it is possible to choose bioactive substances or natural polymers to cover different cosmetic needs and different functions: anti-aging, moisturising, stain-proof, antiseborrhoeic, anticomedogenic, purifying, antioxidant, redensifying, protective, soothing, nourishing, etc. The SCF formulations object of the invention include natural extracts such as for example the extract of the fruits of Prunus spinosa L. (sloe berry, abbreviated PS) for an anticomedogenic gel and the extract of Glycyrrhiza glabra L. (liquorice, abbreviated GG) for an antioxidant emulgel. Further extracts used in the context of the invention are rosehip and rose centifolia extract, lavender, helichrysum, rosemary, grape extract.

[0015] It is therefore an object of the invention a solid cosmetic composition comprising: a. Phospholipid nanoparticles comprising a product obtained by hydroalcoholic extraction or with biosolvents of a plant matrix; and b. a natural film-forming polymer preferably selected from hyaluronic acid, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof; and optionally c. a plasticizing agent, preferably glycerin; wherein said composition is anhydrous, wherein said composition is preservative-free, wherein said composition is in film form, wherein said phospholipid nanoparticles are e.g. ethosomes, phytosomes, liposomes, wherein said composition is reconstitutable into a fluid cosmetic by addition of water upon application to the skin.

[0016] The composition of the present invention is reconstitutable into a fluid cosmetic by adding water in a volume amount of less than or equal to 0.15 mL per square cm of film .

[0017] The film composition is reconstituted in fluid cosmetic in less than 30" from the time of water addition.

[0018] Preferably the phospholipid nanoparticles comprise phospholipids in the outer coating, preferably said nanoparticles are nanovesicles whose outer coating comprises soya lecithin. Preferably the plant matrix used for the preparation of the extract consists of fruits or plants or parts thereof (leaves, flowers, fruits, seeds, bark, rhizome, root), preferably said plant matrix is selected from sloe berry, grape, liquorice, rosehip, centifolia rose, lavender, helichrysum, elderberry, pomegranate, yarrow, bergamot, mint, thyme, rosemary, oregano.

[0019] Preferably, the solid cosmetic composition of the invention further comprises a vegetable oil, preferably selected from almond oil, olive oil, grape seed oil, coconut oil, argan oil, jojoba oil, and rosehip oil.

[0020] Preferably the film has a thickness of 30 to 95 pm , more preferably 30 to 93 pm, still more preferably 30 to 60 pm, still more preferably 40 to 50 pm.

[0021] Preferably, the solid cosmetic composition of the invention can be reconstituted into a fluid cosmetic in the form of a gel, phyto-nanoemulgel, emulsion, lotion, or serum.

[0022] A further object of the invention is a method for obtaining a solid cosmetic composition comprising the following steps: a. Extracting a vegetal matrix with a hydroalcoholic mixture or with biosolvents and evaporating the volatile portion to obtain a solid residue; b. Solubilizing the solid residue from step a. in water and treating it under stirring with the solution comprising a phospholipid in water or ethanol to obtain a suspension of phytosomes or of ethosomes, preferably said phospholipid is soy lecithin; c. Adding to suspension obtained from step b. a film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof, and wherein said polymer is in a concentration comprised from 1% and 5% w / v, preferably 2% w / v; d. Optionally adding glycerin in concentration comprised from 1% and 3% w / v; e. Stirring and storing the mixture obtained from step d. for at least 12-24 hours to obtain the formation of a gel; f. Dehydrating the gel obtained from step e. to obtain a solid film.

[0023] Preferably, in the method according to the invention, the solution of soy lecithin in ethanol is added to the solution of solid residue in water at a rate comprised between 1 mL / min to 2 mL / min.

[0024] Preferably in step f. dehydration is carried out by: drying, lyophilization. More preferably in step f. the dehydration is conducted by drying at a temperature ranging from 35 to 50°C, even more preferably at 40°C.

[0025] Preferably, the drying is carried out in heated air or on a heated surface.

[0026] Preferably, the above-defined method further comprises between the step b. and the step c. the addition of a vegetable oil preferably selected from almond oil, olive oil, grapeseed oil, coconut oil, argan oil, jojoba oil, rose hip oil and mixing to obtain a phyto-nanoemulsion.

[0027] A further object of the invention is a solid cosmetic composition comprising: phospholipid nanoparticles comprising a product obtained by hydroalcoholic extraction or with biosolvents of a plant matrix, a natural filming polymer preferably selected from hyaluronic acid, sodium alginate, carrageenan, gum arabic, xanthan gum, pectin or mixtures thereof; and optionally a plasticizing agent, preferably glycerin; wherein said composition is anhydrous, wherein said composition is preservative-free, wherein said composition is in the form of a film, wherein said phospholipid nanoparticles are e.g. ethosomes, phytosomes, liposomes, and wherein said composition is reconstitutable in a fluid cosmetic by addition of water at the time of application to the skin, obtained by a method comprising the following steps: a. Extracting a vegetal matrix with a hydroalcoholic mixture or with biosolvents and evaporating the volatile portion to obtain a solid residue; b. Solubilizing the solid residue from step a. in water and treating it under stirring with a solution of phospholipid in water or ethanol to obtain a suspension of phytosomes or of ethosomes, preferably said phospholipid is soy lecithin; c. Adding to suspension obtained from step b. a film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof, and wherein said polymer is in a concentration comprised from 1% and 5% w / v, preferably 2% w / v; d. Optionally adding glycerin in concentration comprised from 1% and 3% w / v; e. Stirring and storing the mixture obtained from step d. for at least 12-24 hours to obtain the formation of a homogeneous gel; f. Dehydrating the gel obtained from step e. at a temperature between 35 and 50°C, preferably at 40°C to obtain a solid film. A further object of the invention is the cosmetic use of the composition as defined above or obtainable by the method defined above for the care and cleansing of the skin, with a cosmetic action chosen from anti-aging, anti-stain, antioxidant, redensifying, protective, moisturizing, soothing, nourishing, anticomedogenic, antiseborrheic, purifying.

[0028] DETAILED DESCRIPTION OF THE INVENTION

[0029] The object of the present invention is a solid cosmetic composition preferably in solid and reconstitutable film format in a fluid cosmetic by addition of water at the time of application to the skin. The composition object of the invention preferably comprises substances obtained from the valorization of waste, according to a principle of eco-sustainability.

[0030] The composition object of the invention includes: a. phospholipid nanoparticles comprising a product obtained by extraction of a vegetal matrix with a hydroalcoholic mixture or with biosolvents; and b. a natural film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof; wherein said composition is anhydrous, wherein said composition is preservative-free, wherein said composition is in the form of a film wherein said phospholipid nanoparticles are nanovesicles comprising phospholipids in the outer coating, preferably said nanoparticles are ethosomes, phytosomes, liposomes or the like.

[0031] The phospholipids that can be used in the nanoparticles according to the present invention are for example one or more natural or synthetic phospholipids, pure or in a mixture, such as for example soy lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), sphingomyelin and others. Preferably, soybean lecithin was employed in the invention. The fundamental characteristics of the phospholipids for the invention are the ability to form nanovesicles, the high biological compatibility and the absence of toxicity.

[0032] The composition may further comprise a specific plasticizing agent, in a preferred embodiment said plasticizing agent is glycerin. Although not strictly necessary, glycerin is preferably added as it constitutes a common component in compositions for cosmetic use. Preferably the plant matrix used for the preparation of the extract consists of fruits or plants or parts of them, such as for example leaves, flowers, fruits, seeds, bark, rhizome, root. In a preferred embodiment of the invention the plant matrix is selected from sloe berry, grape, liquorice, rosehip, elderberry, pomegranate, bergamot, yarrow, mint, thyme, rosemary, oregano, centifolia rose, lavender, helichrysum but potentially any other plant matrix may be used.

[0033] The solid cosmetic composition of the invention may further include a vegetable oil, preferably selected from almond oil, olive oil, grape seed oil, coconut oil, argan oil, jojoba oil, or rosehip oil. The addition of a vegetable oil results in a solid composition which, when reconstituted, provides a fluid emulsion, in particular a nanoemulsion.

[0034] In a preferred embodiment the solid cosmetic composition object of the invention is in the form of a solid film preferably of thickness between 40 and 50 pm and in turn reconstitutable in a fluid cosmetic in the form of gel, phytonaneemulgel, emulsion, lotion, serum. The film formulation also has advantages in terms of packaging and transport, also responding to the objectives of sustainability that are the object of the invention.

[0035] The composition is reconstitutable into a fluid cosmetic by adding water in a volume amount of less than or equal to 0.15 mL per square cm of film , preferably the amount of water expressed in Volume (mL) per cm2of film is between 0.10 mL of H2O / cm2of film and 0.15 mL of H 2O / cm2of film, which corresponds to 2-3 drops.

[0036] The film composition is reconstituted in fluid cosmetic in a time less than 30 seconds from the time of addition of water, preferably in a time comprised between 20 and 30 seconds, preferably between 21 and 27 seconds.

[0037] The composition object of the invention, in addition to the essential components described above, may also comprise further additives known and commonly used in the field or functional ingredients, i.e. active ingredients with cosmetic action introduced in the composition at functional doses. The composition of the invention has the ability to incorporate and convey said ingredients once applied to the skin, promoting its skin absorption.

[0038] A further object of the invention is a process for the preparation of said solid cosmetic composition comprising or consisting of the following steps: a. Extracting a vegetal matrix with a hydroalcoholic mixture or with biosolvents and evaporating the volatile portion to obtain a solid residue; b. Solubilizing the solid residue from step a. in water and treating it under stirring with the solution comprising a phospholipid in water or ethanol to obtain a suspension of phytosomes or of ethosomes, preferably said phospholipid is soy lecithin; c. Adding to suspension obtained from step b. a film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof, and wherein said polymer is in a concentration comprised from 1% and 5% w / v, preferably 2% w / v, d. Optionally adding glycerin in concentration comprised from 1% and 3% w / v; e. Stirring and storing the mixture obtained from step d. for at least 12-24 hours to obtain the formation of a gel; f. Dehydrating the gel obtained from step e. to obtain a solid film.

[0039] Preferably, in the method of the invention, the solution of soy lecithin in ethanol is added in a controlled manner to the solution of the solid residue in water, preferably at a rate between 1 mL / min and 2 mL / min.

[0040] Preferably the method defined above further comprises between step b. and step c. the addition of a vegetable oil and the mixing to obtain a phytonaneemulsion. Multiple vegetable oils may be used, such as for example almond oil, olive oil, grape seed oil, coconut oil, argan oil, jojoba oil, rosehip oil.

[0041] Preferably in step of the dehydration is carried out by drying, lyophilization.

[0042] Preferably the drying is in hot air or heated plates.

[0043] In a preferred form in step f. dehydration is carried out by drying at a temperature between 35 and 50°C, preferably at 40°C to obtain a solid film.

[0044] A further object of the invention is a solid cosmetic composition comprising: phospholipid nanoparticles comprising a product obtained by hydroalcoholic extraction or with biosolvents of a plant matrix, a natural filming polymer preferably selected from hyaluronic acid, sodium alginate, carrageenan, gum arabic, xanthan gum, pectin or mixtures thereof; and optionally a plasticizing agent, preferably glycerin; wherein said composition is anhydrous, wherein said composition is preservative-free, wherein said composition is in the form of a film, wherein said phospholipid nanoparticles are e.g. ethosomes, phytosomes, liposomes, and wherein said composition is reconstitutable in a fluid cosmetic by addition of water at the time of application to the skin, obtained by a method comprising the following steps: a. Extracting a vegetal matrix with a hydroalcoholic mixture or with biosolvents and evaporating the volatile portion to obtain a solid residue; b. Solubilizing the solid residue from step a. in water and treating it under stirring with a solution of phospholipid in water or ethanol to obtain a suspension of phytosomes or of ethosomes, preferably said phospholipid is soy lecithin; c. Adding to suspension obtained from step b. a film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof, and wherein said polymer is in a concentration comprised from 1% and 5% w / v, preferably 2% w / v; d. Optionally adding glycerin in concentration comprised from 1% and 3% w / v; e. Stirring and storing the mixture obtained from step d. for at least 12-24 hours to obtain the formation of a gel; f. Dehydrating the gel obtained from step e. to obtain a solid film.

[0045] A further object of the invention is the cosmetic use of the composition as defined above or obtainable by the method defined above for the care and cleansing of the skin, with a cosmetic action chosen from anti-aging, anti-stain, antioxidant, redensifying, protective, moisturizing, soothing, nourishing, antiseborrhoeic, anticomedogenic, purifying.

[0046] Definitions

[0047] In the scope of the present invention, by "a sustainable cosmetic film" is meant a composition, preferably free of water, capable of being reconstituted and transformed at the time of application. A sustainable cosmetic film is obtained by drying a fluid formulation, e.g. a gel, a gelled emulsion, a gelled micro / nanoemulsion or in general an emulgel. As used herein, the acronym "SCF" indicates a Sustainable Cosmetic Film. Alternatively, the term "film" is used in the invention to mean the water-free composition formulated into a solid thin layer.

[0048] In the scope of the present invention, waterless product means an anhydrous product that is free of water. Said product may also be referred to as a solid cosmetic.

[0049] In the scope of the present invention, "ethosomes" means phospholipid nanovesicles, mainly composed of one or more concentric layers of phospholipids, with a relatively high concentration of ethanol (20-45%), glycols and water, for the cutaneous and transdermal delivery of bioactive substances.

[0050] In the context of the present invention, "an ethosomal suspension" means a suspension containing ethosomes dispersed in an aqueous and / or hydroalcoholic matrix.

[0051] In the context of the present invention, "phytosomes (PHY)" means vesicular systems for the release of bioactive substances consisting of phospholipids, in particular lecithin, and functional ingredients of natural origin. Phytosomes are vesicles in which the active agent / bioactive substance is anchored to the polar head of the phospholipid and becomes an integral part of the phospholipid membrane, unlike liposomes, in which the active agent / bioactive substance is generally contained either within the aqueous core or between the hydrophobic tails of the phospholipids of the liposome.

[0052] In the context of the present invention, by "nanoemulsions (NE)" are meant oil-in-water emulsions in which the average diameters of the dispersed droplets are of nanometric scale (from 50 to 1000 nm).

[0053] In the scope of the present invention, by "a phyto-derivative" is meant a pharmaceutical or nutraceutical product composed of ingredients based on plant extracts.

[0054] In the context of the present invention, "phyto-nanoemulsions (PNE)" means nanoemulsions composed of phyto-derivatives.

[0055] In the context of the present invention, "phyto-nanoemulgel (PNEG)" means nanoemulsions obtained from phyto-derivatives and gelled with the addition of natural polymers.

[0056] In the context of the present invention, "a natural polymer" means a material widely present in nature obtained from living cells, resulting for this reason eco-sustainable, biodegradable, biotolerable. Said polymer is chosen e.g. from sodium hyaluronate (HA), Solagum-AX ® (SG; commercial product consisting of a mixture of gum arabic and xanthan gum), sodium alginate (AL), pectin (PA), carrageenan-l (CA). Said polymer upon drying imparts mechanical strength to the formulation, thus it is also referred to as a filming polymer.

[0057] In the context of this invention, preservatives are preferably synthetic preservatives or "substances intended exclusively or mainly to inhibit the development of the microorganism in the cosmetic product" (Article 2 of EU Regulation no. 1223 / 2009). Among the most used in cosmetics are: phenoxyethanol, benzoic acid derivatives, sorbic acid, benzyl alcohol, methyl paraben and propyl paraben listed in Annex V of EU Regulation no.1223 / 2009. These preservatives are used in cosmetology in compositions that contain water; in fact, the more a cosmetic is rich in water, the more it is subject to contamination by microorganisms and said contamination can occur from the production phases to the phase of use of the cosmetic.

[0058] Within the scope of the present invention, "a plasticizing agent" means a substance that, when added to a polymeric material, implements certain characteristics, such as flexibility, workability and deformability. Said plasticizing agent may be e.g. glycerin (gly) and lecithin (LC).

[0059] In the context of the present invention, "dry substance" means the residual part of the ethosomal gel or phyto-nanoemulgel after removal of water at 40°C until a constant weight is reached.

[0060] In the context of the present invention, "biosolvents" are understood to be derived from renewable raw materials, for example from biomass; therefore, they are products of natural origin derived from non-toxic and biodegradable raw materials.

[0061] Within the scope of the present invention, "a plant extract" or "phyto-extract" means a preparation obtained by the evaporation, e.g. partially or totally, of plant juices or solutions resulting from the treatment of a plant substance with a liquid (e.g. water, alcohol, ether) capable of dissolving, and then extracting, the active ingredients. The parts used may be leaves, flowers, fruits, seeds, bark, rhizome, root, or whole plant.

[0062] Within the scope of the present invention, by dehydration stage is meant any process aimed at removing water from the composition such as for example hot air or heated plate drying, freeze- drying. .

[0063] The invention will now be illustrated by means of examples referring to the following figures, given by way of non-limiting example, in which:

[0064] Figure 1. Reduction of reactive oxygen species (ROS) by sloe berry extract solution at different concentrations after A. H2O2 exposure of HaCaT cells and B. UVB radiation exposure of HaCaT cells. Data are expressed as the means of at least nine replicate experiments ± SD. Significance: (*) p <0.05 as compared to positive control cells (ascorbic acid).

[0065] Figure 2. A. FT-IR spectrum of the sloe berry extract. B. Spectrum1H-NMR of the sloe berry extract.

[0066] Figure 3. A. Size of the loaded and unloaded ethosomes during 20 weeks of storage at 4°C. B. The release profile of the polyphenols from the sloe berry extract. (PS) and the encapsulated sloe berry extract. (ETLCPS). C. MTT assays on WS1 cells treated for 24 hours with increasing concentrations of ETLC and ETLCPS. The effects were tested in comparison to control cells. Data are expressed as the means of at least six replicate experiments ± SD

[0067] Figure 4. Viscosity as a function of angular deformation rate for ethosomal gels.

[0068] Figure 5. A. Ethosomal gels before dehydration. B. SCF obtained from ethosomal gels. C. HA2% ETLCPS f. D. SG2%ETLCPS f. E. AL2%ETLCPS f. F. CA1%ETLCPS f.

[0069] Figure 6. Laser scanning confocal microscope images of loaded and unloaded films. A. HA2% ETLCPS f. B. HA2% f.

[0070] Figure 7. Viscosity as a function of the angular deformation rate of the re-dispersed loaded and unloaded gels.

[0071] Figure 8. Modules G' and G" as a function of the frequency of the redispersed gels.

[0072] Figure 9. Antioxidant activity of GG extract and ascorbic acid (AA). Data are expressed as the means of three replicate experiments ± SD.

[0073] Figure 10. Reduction of reactive oxygen species (ROS) by liquorice extract solution at different concentrations after A. H2O2 exposure of HaCaT cells and B. UVB radiation exposure of HaCaT cells. Data are expressed as the means of at least eight replicate experiments ± SD. Significance: (*)p <0.05 as compared to positive control cells (ascorbic acid).

[0074] Figure 11. Viscosity as a function of the angular deformation rate of the PNEGs.

[0075] Figure 12. A. Modulus G' and G " as a function of frequency for the loaded PNEG formulation. B. Modulus G' and G" as a function of the frequency for the unloaded PNEG formulation.

[0076] Figure 13. Flow-curve of the loaded and unloaded PNEGs.

[0077] Figure 14. In Vitro Release of Total Phenol Content (TPC) in PBS: EtOH (7:3 v / v) from extract solution GG (1 mg / mL) and PNEG. Data expressed as the average ± SD (n = 3)

[0078] Figure 15. SCF obtained from PNEG PA loa (phyto-nanoemulgel containing liquorice extract).

[0079] Figure 16. MTT assays on WS1 cells treated for 24 h with increasing concentration of PHYu and PHY. Six replicates were performed for each concentration.

[0080] EXAMPLES EXAMPLE 1: Sustainable cosmetic films for gel products

[0081] 1. Materials

[0082] A hydroalcoholic extract obtained from frozen sloe berries was used. Methanol (MeOH) and ethanol (EtOH) were purchased from Sigma-Aldrich (Milan, Italy). Sodium hyaluronate (CAS: 9067- 32-7) and sodium alginate (CAS: 9005-38-3) were purchased from Farmalabor (Canosa di Puglia, Italy). The Solagum AX® was supplied free of charge by Seppic SA, La Garenne-Colombes (France). Folin-Ciocalteu reagent was kindly provided by Titolchimica (Pontecchio Polesine, Italy). Soybean lecithin (LC) was purchased from Carlo Erba (Milan, Italy). All other chemicals were purchased from Sigma-Aldrich (Milan, Italy). Ultrapure water (18.2 MO cm) was obtained with a MilliQ. apparatus from Millipore (Milford, MA, USA).

[0083] 2. Methods

[0084] 2.1. Preparation and Characterization of Sloe Berry Extract.

[0085] For the hydroalcoholic extraction of the sloe berries the fruits were stripped of seeds and their pulp was homogenized; 10 g of pulp was placed in a 50:50 v / v ethanol / water mixture (150 mL) and sonicated (Transonic TP690 from Elma, Singen, Germany) for 40 minutes at room temperature. The mixture was centrifuged at 4000 rpm for 20 minutes and the supernatants were filtered with filter paper using a Buchner (Rotofix 32A from Hettich, Tuttlingen, Germany). The residue was further extracted by repeating the above-mentioned procedure. Supernatants were collected and evaporated under vacuum at 40°C (Rotavapor Heating Bath B-490) to remove ethanol and most of the water. The concentrated extracts were lyophilized at -48°C for 48 hours and stored at +4.0 ± 1.0°C in the dark until used. Phytochemical characterization tests were performed on the obtained sloe berry extract (PS), including total phenolic content (TPC) determination using the Folin- Ciocalteu reagent, total flavonoid content (TFC) determination using aluminum chloride, antioxidant activity determination with the DPPH assay, LC-DAD-MS / MS analysis of major polyphenols, and the recording of the FT-IR and1H-NMR spectra (for the detailed method of these analysis methods see Sallustio, V.; Chiocchio, I.; Mandrone, M.; Cirrincione, M.; Protti, M.; Farruggia, G.; Abruzzo, A.; Luppi, B.; Bigucci, F.; Mercolini, L.; et al. Extraction, Encapsulation into Lipid Vesicular Systems, and Biological Activity of Rosa canina L. Bioactive Compounds for Dermocosmetic Use. Molecules 2022, 27, 3025. https: / / doi.org / 10.3390 / molecules27093025.).

[0086] A similar procedure can be applied for the preparation of extracts from different natural matrices, both fruits and leaves and roots, such as, for example, rosehip, grape, liquorice, elderberry, pomegranate, bergamot, yarrow, mint, thyme, rosemary, oregano, etc. 2.2. Preparation Of The Ethosomal Suspension

[0087] The ethosomes were prepared using the ethanol injection-sonication method. In particular, soy lecithin (100 mg) was solubilized in ethanol (3 mL) in a covered beaker to avoid ethanol evaporation. The sloe berry extract (10 mg) was solubilized in bidistilled water (7 mL) and mixed uniformly with a magnetic stirrer. Ethanol lecithin solution was added slowly (1 mL / min) to the aqueous solution with a syringe under constant stirring at 700 rpm. The resulting vesicle suspension was homogenized with an ultrasonic bath (Transonic T310, Elma, Germany) for 15 minutes. Unloaded ethosomes, without the addition of sloe berry extract, were prepared as a control.

[0088] The ethosomal suspension containing sloe berry extract (ETLCPS) and the unloaded ethosomal suspension (ETLC) were characterized from a chemical-physical point of view; in addition, their stability and in vitro release were evaluated and cell viability tests were performed to test their safety (see Sallustio, V.; Chiocchio, I.; Mandrone, M.; Cirrincione, M.; Protti, M.; Farruggia, G.; Abruzzo, A.; Luppi, B.; Bigucci, F.; Mercolini, L.; et al. Extraction, Encapsulation into Lipid Vesicular Systems, and Biological Activity of Rosa canina L. Bioactive Compounds for Dermocosmetic Use. Molecules 2022, 27, 3025. https: / / doi.org / 10.3390 / molecules27093025).

[0089] 2.3. Preparation of Natural Polymer-Based Gels

[0090] Different percentages of natural polymers, i.e., sodium hyaluronate (HA), Solagum-AX ® (SG), sodium alginate (AL), carrageenan-l (CA), and glycerin (gly) as a plasticizing agent, were added to 20 mL of ethosomal suspension of natural extract (ETLCPS). Preliminary studies were performed to select the optimal percentage of polymer and glycerin for film preparation. Table 1 shows the composition of the mixtures of ethosomal gels and polymers used for the preparation of the films. Gels obtained from unloaded films (not containing ethosomes and extract) were also prepared as a control using distilled water (20 mL) in place of the ethosomal suspensions and adding the same percentages of polymers and glycerin.

[0091] Table 1. Composition of the mixtures used for the preparation of the films The polymers were slowly added to the ethosomal suspensions. The mixtures were stirred for 24 hours at 300 rpm and stored for 24 hours at room temperature to facilitate gel formation.

[0092] 2.4. Chemical-Physical Characterization of Gels

[0093] 2.4.1. Measurement of pH and viscosity

[0094] The pH of the gels was measured by a digital pH meter, and the viscosity by a rotational viscometer using spindle # 10 for all gels except the alginate gels for which spindle # 11 was used. The measurements were carried out at room temperature.

[0095] 2.4.2. Size determination, polydispersity index (PDI) and zeta potential of vesicular systems

[0096] Size and PDIs were measured by PCS (photon correlation spectroscopy) using the Brookhaven 90- PLUS instrument (Brookhaven Instruments Corp., Holtsville, NY, USA) with a He-Ne laser beam at a wave length of 532 nm (90° diffusion angle). The samples were diluted (1:800 v / v) in ultrapure water. The measurements were performed at room temperature with five runs for each determination. Zeta potential measurements were performed at 25°C with the Malvern Zetasizer 3000 HS instrument (Malvern Panalytical Ltd., Malvern, UK), using the same dilution conditions as above.

[0097] 2.5. Film Preparation from Gels

[0098] Approximately 5 g of each mixture was weighed in a Petri dish (diameter 57 mm and height 10 mm) and oven-dried at 40°C for 4 hours (FD series oven, Binder, Tuttlingen, Germany). Preliminary tests were performed to determine the optimal drying time by weighing the blends in the Petri dishes until a constant weight was reached. After 4 hours drying in the oven, the Petri dishes were weighed again to determine the dry substance. The films were then gently removed from the supports and stored in a polyethylene bag and maintained in a silica gel desiccator to prevent moisture absorption until further analysis.

[0099] 2.6. Chemical-Physical Characterization of the Films

[0100] 2.6.1. Determination of dry matter

[0101] The dry matter values were obtained from the difference between the weight of the gel in the Petri dishes and the weight of the film after drying. The measurements were performed in triplicate.

[0102] 2.6.2. Thickness Determination The thickness of the films was measured at five different positions taken randomly with a digital gauge with an accuracy of 0.01 mm. Three films were measured by formulation.

[0103] 2.6.3. Macroscopic Appearance and Determination of Film Transparency by Uv-Vis Spectroscopy

[0104] The macroscopic appearance of the film was observed by visual and tactile inspection to evaluate the homogeneity of the appearance, the consistency and the ease of removal from the support. The transparency of the film was observed spectrophotometrically with UV-Vis 1601 spectrophotometer (Shimadzu). In particular, 1x2 cm2film strips were placed in a cell of the spectrophotometer and the transmittance spectrum was recorded between 200-800 nm. The transmittance was related to the area under the curve, the greater the area, the greater the transmittance. Measurements were made on three film samples for each formulation.

[0105] 2.6.4. Laser Scanning Confocal Microscopy

[0106] Confocal laser scanning microscopy (CLSM) is a technique for imaging samples containing fluorophores. High-resolution images are generated by the superposition of photons emitted by the fluorophore that reach the detector during an exposure period. A 1 cm2square of each type of film was analysed using a Nikon Cl Laser Scanning Confocal Microscope, equipped with Nikon PlanApo 40, 1.4-NA Oil Immersion Lens. Excitement was performed at 405-488-543 nm with an argon laser and emission was recorded at 450 / 35-515 / 30-650 nm. Images were analysed by Image J Software (version 1.53a, U.S. National Institutes of Health, Bethesda, MD, USA).

[0107] 2.6.5. Determination of Total Phenolic Content (TPC) and Antioxidant Activity (AA%)

[0108] From each film were cut out squares with a surface area of 1.20 cm2, to which was added 2 mL of distilled water and mixed until gel formation. Subsequently, 0.2 mL of gel was analysed for TPC by the Folin Ciocalteu reagent using the procedure described by Singleton et al. (Singleton et al., 1999). Specifically, 0.2 mL of the gelled solution was added to 1 mL of 1:10 diluted Folin-Ciocalteu reagent, followed by addition of 0.8 mL of sodium carbonate solution (7.5% w / v). After 25 minutes in the dark at 40.0±1.0°C, samples were centrifuged at 12000 rpm for 5 minutes and absorbance at 750 nm was measured by spectrophotometry (UV-Vis 1601 spectrophotometer, Shimadzu). Distilled water was used as white. The TPC was calculated from a gallic acid standard curve (R2=0.999). All measurements were performed in triplicate and the results were expressed as micrograms of TPC per square cm of film (pg / cm2).

[0109] 2.6.6. Determination of Antioxidant Activity A 1 mL aliquot of the redispersed gels (obtained as in paragraph 2.6.5.) was taken and added with 1 mL of a solution of 2,2'-di-phenyl-l-picrylhydrazyl (DPPH) (0.1 mM in methanol) to determine the residual antioxidant activity of the 2,2' -di-phenyl-l-picrylhydrazyl radical (DPPH), as described by Brand-Williams et al. (Brand-Williams et al., 1995) with minor modifications. Specifically, 1 mL of each gel was mixed with 1 mL of DPPH solution (0.1 mM in methanol) at room temperature. The mixtures were kept in the dark for 25 minutes, centrifuged at 1200 rpm for 5 minutes and the absorbance was measured at 517 nm. The methanol was used as blank and the DPPH solution was used as control. The results were expressed as a inhibition percentage of the DPPH radical according to the following equation: Inhibition (%) = [(Ao -A) / Ao] where Ao is the absorbance of the DPPH control and A is the absorbance of the sample spiked with DPPH. The test was done in triplicate for each type of film.

[0110] 2.7. Determination of Film Dissolution Time and Minimum Amount of Water Required For Rehydration

[0111] A 1.2 cm2square of each film type was placed in a 10 mL beaker and 2 mL of distilled water at 32°C was added to simulate skin temperature. The suspension was stirred at 100 rpm and the time to complete dissolution was measured by visual inspection. Then 5 pL of the gels, obtained from the films thus redispersed, were diluted 1:800 (v / v) with ultrapure water and the size and polydispersity index of the vesicular systems were measured by pcs (photon correlation spectroscopy) as reported in paragraph 2.4.2.

[0112] Since rapid gel formation with a minimum amount of water is crucial for this type of film formulation, the following additional tests were performed for the most promising films. A sample of the selected films of square shape and surface of 1 cm2was placed on a glass surface and distilled water was added dropwise until a homogeneous transparent or translucent gel was obtained, thereby determining the minimum amount of water necessary for gelation.

[0113] Subsequently a similar sample was placed on a glass surface, the minimum amount of water necessary for gelation was added and the time for complete rehydration of the film and the formation of the homogeneous gel was timed. Verification of the formation of a homogeneous gel was obtained by macroscopic visual observation of transparency / translucency. Tests were performed at 20±l°C and in triplicate.

[0114] 2.8. Rheological Measurements of Redispersed Films (fr)

[0115] Rheological studies were carried out on the redispersed films. Analyses were performed with the Malvern Kinexus Rheometer Lab+ rheometer (Malvern Instruments, UK) using cone-plate geometry. Viscosity measurements were made between 0.1 and 10 s’1. All measurements were performed at 25.0 ± 0.2°C. Viscosity values (Pa-s) were reported as a function of angular deformation rate.

[0116] The viscoelastic properties of the fluid gels produced were measured by small amplitude oscillatory experiments with a fixed deformation of 2%, value within the linear region, using a frequency range between 0.1 and 10 Hz. The measurements were performed at 25.0 ± 0.2°C.

[0117] The stickiness (responsible for the unpleasant sticky feeling of a cosmetic) and the adhesiveness (the ability of a cosmetic to adhere to the skin over time) were measured for gels using the same equipment (Malvern Kinexus Rheometer Lab+) with the plate-plate geometry (pull-away test). An instrument kit with conditions of 0.1 mm / s, 5 mm and 0.15 gap was selected. In this test, the peak negative normal force required to separate the two parallel plates holding the gel can be attributed to stickiness. The area under the force-time curve is instead related to adhesiveness. Six measurements were performed for each sample.

[0118] 2.6. Measurement of the Spreadability of Redispersed Films

[0119] A parallel plate method was used to measure the spreadability of the redispersed films. Gel samples obtained from the redispersed films (300 pL) were distributed between two sheets of Sil-Tec 304 mm x 304 mm x 0.127 mm medical grade solid silicone rubber (Technical Products Inc. of GA, USA). A weight of 200 g (20 cm diameter) was applied to it for 30 seconds. The diameter of the gel after application of the weight was measured. Measurements were performed at room temperature (25 ± 2°C). Spreadability in mm2was calculated with formula: =2x / 4, where S is the area (mm2) and d is the diameter (mm) of the spreadability area.

[0120] 3. Results

[0121] Based on previous studies on ethosomes containing natural extracts, it was thought to use these formulations as a cosmetic ingredient to prepare a new waterless cosmetic formulation or natural polymer films containing natural extracts encapsulated in phospholipid vesicles called sustainable cosmetic films (SCF). The formulation was developed starting from a sloe berry hydroalcoholic extract encapsulated in ethosomes. Subsequently the ethosomal suspension was gelled using natural polymers; following low temperature dehydration of the ethosomal gels the films named SCF were obtained.

[0122] 3.1. Phytochemical Characterization of Sloe Berry Extract The sloe berry extract has a total polyphenol content determined by the Folin-Ciocalteu method equal to 13.99±0.04 pg GAE / mg of extract where GAE indicates the equivalents of gallic acid.

[0123] The total content of flavonoids obtained by the aluminium chloride method is 3.14±0.15 pg QE / mg extract where with QE the equivalents of quercetin are indicated.

[0124] The antioxidant activity of a lmg / mL extract solution measured as DPPH-lowering capacity was 91.78±0.80%.

[0125] This value is in agreement with the values of antioxidant activity measured as reduction of intracellular ROS stimulated by H2O2 or by UV in HaCaT cells as reported respectively in Figure 1A and IB towards ascorbic acid (AA) control.

[0126] In the FT-IR spectrum (Figure 2A) it is possible to recognize the main functional groups of the substances present in the extract. This FT-IR spectrum of the sloe berry extract is similar to those already reported in the literature (Andronie et al., 2019; Magiera et al., 2022).

[0127] 1H-NMR analysis of the sloe berry extract (Figure 2B) revealed a high amount of sugars, particularly glucose as evidenced by the presence of the 6 5.2 and 6 4.6 duplexes attributable to the anomeric protons of glucose a and p, respectively. Another primary metabolite detected is aspartic acid. In the anomeric region (between about 6 8 and 6) the presence of signals characteristic of caffeic acid is evident (Figure 2B enlarged region).

[0128] Finally, LC-DAD-MS / MS analysis determined the amounts of some bioactive substances belonging to different classes of polyphenols and ascorbic acid. The values obtained are reported in Table 2.

[0129] Table 2. Content of polyphenols and ascorbic acid in sloe berry extract *

[0130] *Values expressed as the average ± standard deviation (n=3)

[0131] 3.2. Chemical-Physical Characterization og the Sloe Berry Ethosomes

[0132] The results on the extraction of hookworm show a high amount of bioactive compounds. However, their biological properties could be lost due to many factors, such as time, oxidation and light exposure. For this reason, to overcome these drawbacks, the sloe berry extract was encapsulated in phospholipid nanovesicles. Several types of vesicles have been tested in preliminary studies (liposomes, hyalurosomes, ethosomes). The formulation in which the sloe berry extract was encapsulated in lecithin ethosomes (ETLCPS) was the most promising for the preparation of a redispersible film for a topical gel. The chemical-physical characterization of the ethosomes containing sloe berry extracts and of the drained ethosomes prepared as a control is reported in Table 3.

[0133] Table 3. Size (nm), polydispersity index (PDI), zeta potential (mV), and encapsulation efficiency (EE%) of ethosomes loaded with sloe berry extract (ETLCPS) and unloaded (ETLC).*

[0134] *Values expressed as the average ± standard deviation (n=3)

[0135] Stability studies have shown that both loaded and unloaded ethosomes maintain similar size over time to the initial ones over a period of 20 weeks at 4°C as evidenced by Figure 3A.

[0136] In vitro release studies (Figure 3B) show that ethosome-encapsulated extract is released more slowly than free extract and this may favour gradual absorption into the skin.

[0137] Finally, cell viability studies were conducted with the MTT assay to verify the safety of the ethosomal suspension and, as shown in Figure 3C, the sloe berry ethosomal suspension was found to maintain cell viability even when the highest concentrations were used.

[0138] 3.3. Characterization of Ethosomal Gel for SCF

[0139] The pH of the ethosomal gels, the size, the PDI and the zeta potential of the ethosomal vesicles were evaluated after 24 hours from their preparation. The data are reported in Table 4.

[0140] Table 4. PH value, size (nm) and PDI and zeta potential of ethosomal gels with sloe berry extract and unloaded gels

[0141] *Values are expressed as the average ± SD, (n = 3) All loaded gels, i.e. containing sloe berry extract, have a pH of 5.5, that is optimal for topical use. The pH of the ethosomal gels with the extract was lower than that of the drained gels, where the values range from 6.3 to 7. The lowering of the pH is due to the organic acids in the sloe berry extract, mainly malic, ascorbic, aspartic, coumaric and caffeic acid. Regarding the size, the loaded gels maintained nanometric values for the vesicles, except for the gel with carrageenan. At the same time, the unloaded gels had larger nanoparticles, probably due to an inhomogeneous aggregation of the polymer chains, as also confirmed by the higher PDI values. Among the loaded gels, the alginate ethosomal gel showed a lower value for size and PDI (313.75 ±1.20 nm and 0.264±0.003, respectively). In contrast, Solagum AX® ethosomal gel showed higher values in the following parameters: 322.93±24.48 nm for size and 0.358±0.030 for PDI. Finally, the zeta potential values of the loaded and unloaded formulations are all negative, highlighting that all the polymers considered have negative charges. However, a further decrease in zeta potential can be observed for the filled formulations, ranging from -58.39 ±13.53 SG2% ETLCPS to -67.37±3.57 HA2% ETLCPS. These results indicate that the lecithin and the extracted compounds contribute to the negative final value of the zeta potentials and help prevent the aggregation of the nanoparticles by increasing their stability over time. Importantly, the zeta potential of ETLCPS (-30.92 ±1.02) is strongly decreased by the added polymers used in ethosomal gels, and this could be due to the fact that the natural polymer forms a coating on the surface of the vesicles, as also observed by Xie et al. (Xie et al., 2018).

[0142] 3.4. Viscosity as a function of the angular deformation rates of the ethosomal gels for SCFs

[0143] The rheological behaviour of the prepared mixtures was evaluated by plotting the viscosity value as a function of the rate of angular deformation ("shear rate"), as shown in Figure 4. All mixtures can be classified as non-Newtonian fluids because the viscosity decreases as the angular deformation rate increases.

[0144] It can also be seen from Figure 4 that the alginate ethosomal gels have high values of viscosity. Another consideration is that generally the loaded ethosomal gels showed higher viscosity values than the corresponding unloaded gel. The increase in viscosity is probably attributable to the presence of lecithin ethosomes.

[0145] 3.5. Chemical-Physical Characterization of the Films

[0146] 3.5.1. Macroscopic appearance

[0147] The films were obtained from the ethosomal gels following the procedure described in paragraph 2.3. After preparation, the films were removed from the Petri dishes and observed prior to storage in the dryer. The films were observed macroscopically by visual and tactile inspection and their macroscopic appearance is shown in Figure 5. All loaded films exhibit optimal macroscopic characteristics, i.e. homogeneity, absence of rips and are relatively transparent, indicating that the vesicles are uniformly incorporated into the polymer matrix. Regarding the loaded films, HA2% ETLCPS f is easy to remove from the support and the appearance is translucent; SG2% ETLCPS f requires a gentle removal procedure and is almost transparent; AL2% ETLCPS f is easier to remove and translucent in appearance; CA1% ETLCPS f is too fragile and thin and it was impossible to obtain an intact film by removal; for this reason it was not considered in some of the subsequent characterizations. Likewise, the unloaded films of SG and CA were sticky, difficult to detach from the Petri dishes, and form folds with ease, highlighting that the presence of the phospholipid vesicles confers further strength and flexibility to the film.

[0148] 3.5.2. Determination of transparency by UV-Vis transmittance

[0149] The transparency of the films was also determined spectrophotometrically from the values of the areas under the % transmittance curves, as reported in Table 5.

[0150] Table 5. Area-under-the-curve values for Transmittance % of Films

[0151] *Values are expressed as the average ± SD, (n = 6)

[0152] The highest transmittance value was observed for the HA2% unloaded film; the ability of hyaluronic acid to form highly transparent films is known. In contrast, the film loaded with HA2% has a greater opacity than the unloaded film, highlighting the homogeneous distribution of ethosomes. Surprisingly, for the Solagum-AX ® polymer, the transparency of the loaded film is greater than that of the unloaded film.

[0153] 3.5.3. Determination of the dry substance and thickness of the loaded and unloaded films

[0154] The dry substance was determined by the difference between the weight of the empty Petri dishes and the weight after film formation. The values are reported in Table 6. The difference in dry matter values could be attributed to the different composition of the ethosomal gels before their evaporation to obtain the films. Overall, the loaded films showed a higher dry substance than the unloaded films. Table 6. Dry matter value (%) and thickness (pm) for loaded and unloaded films

[0155] *Values are expressed as the average ± SD, (n = 3)

[0156] The thickness of the film depends on the method of preparation, the amount of gel poured and the flatness of the drying surface (Rezvanian et al., 2016). The average thicknesses of the films are reported in Table 6. As with the dry matter values, the thickness is greater for the filled formulations. In particular, the loaded alginate showed the highest thickness value (93.3±2.19 pm). The thickness of the loaded films follows the order AL2% ETLCPS f > HA2% ETLCPS f > SG2% ETLCPS f > CA1% ETLCPS f. The order listed for the thickness reflects the ease of detaching the films from the Petri dish. Thus, this is a significant parameter even for the selection of the most easily scalable formulation.

[0157] 3.4.4. Determination of TPC and AA of the Films Loaded with Extract

[0158] The TPC content and AA of the loaded films were examined and the results are tabulated in Table 7.

[0159] The TPC and AA values of the films follow the following order AL2% ETLCPS f > SG2% ETLCPS f> HA2% ETLCPS f, confirming that the antioxidant activity is mainly due to polyphenols as bioactive compounds. Importantly, this method of preparation allows films containing encapsulated extracts to be obtained while retaining their antioxidant activity; SCFs are therefore effective formulations for preserving the benefits of bioactive compounds.

[0160] Table 7. Determination of TPC (pg / cm2) and AA% of loaded films

[0161] *Values are expressed as the average ± SD, (n = 3)

[0162] 3.4.5. Analysis of Mechanical Properties of Loaded Films

[0163] The tensile test results of the loaded films are shown in Table 8.

[0164] Table 8. Tensile Test Results of Loaded Films.

[0165] *Values are expressed as the average ± SD, (n = 3)

[0166] Tensile test values are influenced by the presence of lecithin and glycerin as plasticizers that promote the formation of hydrogen bonds. However, natural polymers also play a role in the tensile stress response and, for the parameters considered, the order is the following HA2% ETLCPS f > SG2% ETLCPS f > AL2% ETLCPS f.

[0167] 3.4.6. Laser Scanning Confocal Microscopy

[0168] Laser scanning confocal microscopy was used to highlight the presence and distribution of ethosomes in the loaded films. As shown by way of example in Figure 6, the HA2% loaded film shows the presence of a homogeneous dispersion of ethosomes with respect to the corresponding unloaded film.

[0169] 3.5. Film Redispersion and Determination of Minimum Amount of Water for Rehydration

[0170] SCFs are designed as a waterless formulation to be rehydrated when applied to the skin. For this reason, the times necessary to obtain homogeneous gels from the films redispersed in water (fr) under conditions similar to those of the final use were measured and the results are shown in Table 9a

[0171] Table 9a Redispersion time (in seconds) of films in water

[0172] *Values are expressed as the average ± SD, (n = 3)

[0173] For loaded films, the redispersion in water follows the orderHA2% ETLCPS fr < AL2% ETLCPS fr < SG2% ETLCPS fr < CA1% ETLCPS fr. A similar order was found for the unloaded films, indicating that this parameter depends mainly on the type of polymer used. In conclusion, considering that a rapid redispersion could be beneficial for users, HA2% ETLCPS f has optimal characteristics for this sustainable cosmetic approach.

[0174] In particular, for these films that showed the best redispersibility, the minimum amount of water and the minimum value of time necessary for rehydration were determined to obtain a homogeneous gel.

[0175] The results are shown in Table 9b.

[0176] Table 9.b Minimum amount of water and minimum rehydration time

[0177] *Minimum amount of water for the formation of a homogeneous gel

[0178] ** Minimum time required for rehydration of 1 cm2of film with 0.15mL of H20

[0179] 3.6. Physicochemical Characterization of Redispersed Films

[0180] After redispersion of the films, the size and PDI of the nanoparticles were measured to verify that their nanometric structures and homogeneity were maintained. The results are shown in Table 10.

[0181] Table 10. Vesicle size (nm) and PDI of loaded and unloaded films after redispersion in water

[0182] *Values are expressed as the average ± SD, (n = 3)

[0183] As reported in Table 10, the HA2% ETLCPS fr formulation shows nanoparticles with smaller sizes (707.6±104.2 nm) than the other formulations. Overall all loaded films (except CA1% ETLCPS fr) have nano-size particles after their redispersion in water. 3.7. Rheological Characterization of Redispersed Films

[0184] Figure 7 shows the viscosity trend as a function of the angular deformation rates of the redispersed films. As can be seen in Figure 7 , HA2% ETLCPS fr has a Newtonian behaviour, while SG2% ETLCPS fr shows a non-Newtonian behaviour with a shear-thinning effect, i.e. the viscosity decreases with increasing angular deformation rate. The shear-thinning effect is considered favourable for facilitating the topical application of a product. AL2% ETLCPS fr has viscosity values too low to be represented in the graph.

[0185] Figure 8 shows the trend of elastic modulus G' and viscous modulus G" for the different redispersed SCFs.

[0186] In this oscillation test carried out to evaluate the viscous and elastic modulus of the redispersed films, it can be observed that modulus G' is prevalent for Solagum ®AX formulations while modulus G" has higher values for formulations containing hyaluronic acid and alginate. This result depends on the chemical-physical characteristics of the polymers

[0187] 3.8. Adhesion test

[0188] The adhesion test was performed to evaluate the stickiness and adhesiveness of the redispersed gels and the results are reported in Table 11.

[0189] Table 11. Peak Normal-Force / Normal-Force(N) and Area under curve Force-Time (N-s) for redispersed gels

[0190] *Values are expressed as the average ± SD, (n = 6)

[0191] In Table 11 for Peak Normal-Force / Normal-Force means the peak of normal force necessary to separate the two plates that include the gel and represents an index of the stickiness of the formulation; for Area under curve Force-Time means the overall force required for the detachment of the two plates that include the gel and are therefore an index of the adhesion capacity of the gel. It can be seen that the values obtained are lower for the loaded films. This favourable reduction in stickiness is attributable to the presence of the ethosomes and components of the extract.

[0192] 3.9. Determination of the Spreadability of Redispersed Films

[0193] Spreadability assessment is a useful parameter as it could influence consumer appreciation of the product. Considering the loaded films, Table 12 shows that HA2% ETLCPS fr and SG2% ETLCPS fr have a higher spreadability than the respective unloaded formulation according to the results of the adhesion test. The lower feeling of stickiness and greater spreadability could be due to the presence of lecithin and the substances present in the extract.

[0194] Table 12. Spreadability (mm2) of the re-dispersed loaded and unloaded films.

[0195] *Values are expressed as the average ± SD, (n = 3)

[0196] EXAMPLE 2: Sustainable Cosmetic Films For Phyto-Nanoemulgels

[0197] 1. Materials

[0198] Plant material: the roots of Glycyrrhiza glabra L. were donated by Naturemed s.r.l. (Castrovillari, CS). Methanol (MeOH) and ethanol (EtOH) were purchased from Sigma-Aldrich (Milan, Italy). Sodium hyaluronate (CAS 9067-32-7) and sodium alginate (CAS: 9005-38-3) were purchased from Farmalabor (Canosa di Puglia, Italy). Solagum AX® was kindly provided by Seppic SA, La Garenne- Colombes (France). Folin-Ciocalteu reagent was purchased from Titolchimica (Pontecchio Polesine, Italy). Soybean lecithin (LC) was purchased from Carlo Erba (Milan, Italy). All other chemicals were purchased from Sigma-Aldrich (Milan, Italy). The Phosphate Buffer Solution at pH 7.4 (PBS) was prepared with the following composition: 2.98 g / L NazHPC xl2 H2O, 0.19 g / L KH2PO4, 8 g / L NaCI. Ultrapure water (18.2 MO cm) was obtained with a MilliQ. apparatus from Millipore (Milford, MA, USA).

[0199] 2. Methods 2.1. Extraction and Characterization of Liquorice Extract

[0200] For the extraction of liquorice roots 50 g of raw material were weighed into a flask and added with a 50:50 v / v ethanol / water mixture up to the volume of 500 mL. The mixture was sonicated for 1 h and stirred for three days in the dark. Then, the mixture was filtered and dried in a rotary evaporator. Two additional extractions were performed on the same plant material by repeating the same procedure twice.

[0201] The extract was characterized by determination of total polyphenol content (TPC), total flavonoid content (TFC), antioxidant activity (AA%) by DPPH. The reduction of ROS induced in keratinocytes by F Ozor UVB radiation was also evaluated.

[0202] 2.2. Phytosome preparation (PHY), phyto-nanoemulsion (PNE) and phyto-nanoemulgel (PNEG)

[0203] Preliminary studies were carried out to select the type of vegetable oil and the percentages of the different substances to obtain a nanoemulsion of liquorice extract (GG), reaching the following preparation method. The first step was the preparation of the phytosomes (PHY): in a beaker the GG extract (20 mg) was mixed with soy lecithin (400 mg) and distilled water (20 mL) and stirred at 300 rpm for 2 hours. Then, the phytosome suspension was sonicated for 10 minutes to reduce the size of the phytosomes. After sonication, almond oil (2 mL) was added, followed by ethanol (2 mL) and glycerin (0.2 mL). Subsequently, the mixture was homogenised for 10 minutes to obtain the phyto-nanoemulsion (PNE). After homogenization, several polymers (hyaluronic acid, Solagum AX or a pectin / alginate mixture) were added to the phyto-nanoemulsion as gelling agents to obtain the different types of phyto-nanoemulgels (PNEGs). The prepared formulations were stirred for 24 hours and stored for a further 24 hours to complete the gel preparation before their chemicalphysical and rheological characterization. The composition of the prepared formulations is shown in Table 13.

[0204] Table 13. Preparation of phytosomes (PHY), phyto-nanoemulsions (PNE) and phyto-nanoemulgels (PNEG) *GG-liquorice extract; AO- almond oil; LC-soya lecithin; Gly-Glycerin; HA-hyaluronic acid; SG- Solagum AX; P / A-pectin / alginate mixture;

[0205] 2.3. Chemical-Physical Characterization of PHY, PNE and PNEG

[0206] 2.3.1. Size and Dimensional Distribution

[0207] Nanoparticle sizes of the different preparations were measured by DLS (Dynamic Light Scattering) using Brookhaven 90-PLUS instrument (Brookhaven Instruments Corp., Holtsville, NY, USA). The sample was diluted to 1:2000 (v / v) with ultrapure water. The results were reported as the mean of five consecutive readings.

[0208] 2.3.2. Determination of Zeta Potential

[0209] Zeta potential measurements were performed at 25°C using a Malvern Zetasizer 3000 HS (Malvern Panalytical Ltd., Malvern, UK), employing the same 1:2000 (v / v) dilution with ultrapure water.

[0210] 2.3.3. pH Determination

[0211] The pH values of the formulations were determined using a digital pH meter (Crison Instruments, S.A. Barcelona, Spain) at room temperature and the measurements were performed in triplicate.

[0212] 2.3.4. Determination of TPC and AA content %

[0213] For the determination of TPC content, about 1 g of each type of PNEG formulation was weighed in a beaker and dispersed in 10 mL of methanol, sonicated for 60 minutes and centrifuged at 12,000 rpm for 15 minutes. TPC and AA% of PNEGs were determined by Folin-Ciocalteu method and DPPH test as reported in paragraph 2.6.5.

[0214] 2.3.5. Density Determination

[0215] The density was calculated as the weight to volume ratio of the PNEGs according to the following formula PNEG density = m / V and expressed in g / mL.

[0216] 2.4. Rheological Characterization of PNEG

[0217] Rheological studies were conducted on phyto-nanoemulgels stored at room temperature for 2 days after their production. Analyses were performed with the Malvern Kinexus Rheometer Lab+ instrument (Malvern Instruments, UK) using cone-plate geometry. Viscosity measurements were made between 0.1 and 10 s’1. All measurements were performed at 25.0 ± 0.2°C. Viscosity values (Pa-s) were reported as a function of angular deformation rate.

[0218] The viscoelastic properties of the fluid gels produced were measured by small amplitude oscillatory experiments with a fixed deformation of 2%, within the linear region, using a frequency range between 0.1 and 10 Hz. The measurements were performed at 25.0 ± 0.2°C.

[0219] The stickiness (or tackiness) and adhesiveness of the gels were measured using the same equipment with a plate-plate geometry (pull-away test). An instrument kit with conditions of 0.1 mm / s, 5 mm and 0.15 gap was selected. In this test, the peak negative normal force required to separate the two parallel plates holding the gel can be attributed to stickiness. The area under the force-time curve represents the adhesive force. Six measurements were performed for each sample.

[0220] To measure spreadability, a parallel plate method was used. Samples of phyto-nanoemulgel (300 pL) were spread between two sheets of medical-grade solid silicone rubber Sil-Tec 304 mm x 304 mm x 0.127 mm (Technical Products Inc., GA, USA). A circular weight of 200 g (20 cm in diameter) was applied to it for 30 seconds. Thereafter, the change in diameter was measured. Measurements were performed at room temperature (25 ± 2°C). The spreadability, expressed in mm2, was calculated using the formula: =2x / 4, where S is the area (mm2) and d is a diameter (mm). The measurements were performed in triplicate for each sample.

[0221] 2.5. Stability Studies of PNEG

[0222] All formulations were analysed for macroscopic appearance (visual inspection) after preparation and after 4 and 8 weeks of storage at room temperature (25 ± 2 °C). The nanoparticle size, size distribution and pH of the formulations were determined as described above.

[0223] 2.6. In vitro release studies of PNEG

[0224] In vitro release studies were performed to assess the release of bioactive substances from the matrix. Release through a cellulose membrane was evaluated for these studies and Franz cells were used. In particular, a cellulose membrane was positioned between the donor and recipient compartments of Franz cells to assess the in vitro release of polyphenols from PNEG formulations. 0.5 mL of each sample was poured into a donor chamber and 12 mL of 70 / 30 (v / v) PBS / EtOH mixture was used as delivery medium. 200pL of release medium were withdrawn at predetermined time intervals (1, 2, 3, 4, 5 and 6 hrs) and replaced with the fresh medium. The TPC released in each sample was determined by the Folin-Ciocalteu assay as previously reported. The results were expressed as a percentage of TPC released over time. All experiments were performed in triplicate. 2.7. Preparing films from PNEG

[0225] About 4 g of each mixture was weighed in a Petri dish (diameter 3.5 mm and height 10 mm) and oven-dried (FD series, Binder, Tuttlingen, Germany) at 40°C for 4 hours. Preliminary tests were performed to determine the optimal drying time by weighing the blends in the Petri dishes until a constant weight was reached. After 4 hours drying in the oven, the Petri dishes were weighed again to determine the dry substance. Subsequently the films were gently removed from the media; the thickness of the films was measured using a digital gauge. The films were stored in a polyethylene bag and kept in a desiccator with silica gel to prevent moisture absorption until analysis.

[0226] 3. Results

[0227] 3.1. Extraction of the GG roots and characterization of the liquorice extract.

[0228] GG roots supplied by Naturemed srl (Italy) were extracted from a 50:50 (v / v) ethanol / water mixture followed by vacuum drying. The extraction yield of liquorice root was 28.08%. An extract solution of 1 mg / mL was prepared and analysed; the same concentration was used to prepare vesicle systems for encapsulation of the extract. The pH of this solution measured by a digital pH meter is 6.40±0.02.

[0229] TPC was determined spectrophotometrically using the Folin-Ciocalteu method, and was found to be equal to 62.11±0.41 pgGAE / mg extract (micrograms gallic acid equivalent / mg extract).

[0230] The TFC was determined by a UV-vis spectrophotometric test using aluminum chloride. The TFC value of the GG extract is 6.39±0.13 pg QE / mg extract. In the literature it is reported that almost 300 types of flavonoids are present in the GG extract, in particular liquiritigenin, quercetin and glabridin (Wahab et al., 2021).

[0231] The phenolic compounds present in the GG extract determine its antioxidant activity. In the present study, AA% was determined at different extract concentrations (0.1, 0.25, 0.5, 1 mg / mL) by DPPH assay compared to ascorbic acid used as control. Figure 9 shows that the AA% increases with the concentration of the GG extract and, at 1 mg / mL, reaches the value of 87.38±2.64%. This result is in accordance with the data of the literature and it seems that 1 mg / mL is a suitable concentration for preparing antioxidant cosmetic formulations.

[0232] This result was confirmed by the ROS reduction test which measures the ability to reduce intracellular ROS generation induced by a chemical compound (H2O2) or by UV light in vitro on the human keratinocyte HaCaT cell line. The results are shown in Figures 10A and 10B. Based on these results, it can be concluded that the phenolic compounds in the GG extract have an antioxidant activity applicable in the cosmetic field.

[0233] 3.2. Preparation and Characterization of Phytosomes and Phyto-Nanoemulsion

[0234] Optimized formulations determined from preliminary studies were prepared and characterized as follows:

[0235] Table 14. Size (nm), PDI, ( potential, and pH of loaded phytosomes (PHY); unloaded phytosomes (PHYu), loaded phyto-nanoemulsion (PNE), and unloaded phyto-nanoemulsion (PNEu).*

[0236] *Values are expressed as the average ± SD, (n = 3)

[0237] In Table 14 the main characteristics of phytosomes and phyto-nanoemulsion particles are shown. The phytosome sizes measured are 674.0±34.5 nm. The addition of almond oil, ethanol and glycerin led to the formation of nanoemulsions with particle sizes of 288.0 ± 13.6 nm. All filled formulations are larger than unloaded, probably due to the encapsulation of bioactive compounds in the phospholipid vesicles. The PDI value increased from 0.223±0.040 to 0.263±0.013 for PHY and PNE respectively. However, the PDI remains less than 0.3, indicating a narrow size distribution of the nanoparticles. A decrease in zeta potential value from -51.77 ±0.25 to -58.83±1.27 is observed between PHY and PNE. These values are in both cases highly negative, highlighting a potential stability of the system due to the electrostatic repulsion between the vesicles that could prevent their aggregation. Finally, the pH value of the PNE is slightly acidic (5.33), and is suitable for topical use of the formulation.

[0238] 3.3. Formulation and Characterization of PNEGs

[0239] Table 15. Size (nm), PDI, ( potential, pH, TPC (pg / mL) and AA% of the phyto-nanoemulgels of hyaluronic acid loaded (PNEG HA loa) and unloaded (PNEG HA UNL), Solagum-AX loaded (PNEG SG loa) and unloaded (PNEG SG UNL) and pectin / alginate loaded (PNEG PA loa) and unloaded (PNEG PA UNL).*

[0240] *Values are expressed as the average ± SD, (n = 3)

[0241] Several gelling polymers were added to the nanoemulsion formulation. In fact, the addition of the gelling agent facilitates the application to the skin and the absorption of the bioactive compounds and imparts additional properties to the final formulation due to the specific benefits of the polymer. In detail, hyaluronic acid has anti-aging properties, and Solagum AX® has moisturising properties, properties also common to pectin and alginate. In Table 15 are reported the main parameters for nanoparticles. In particular, with regard to size, the addition of the polymer increases the size of the nanoparticles following the order PNEG PA loa> PNEG SG loa> PNEG HA loa. This increase is probably due to the interaction of the polymer with the surface of the nanoparticles. The PDI value of less than 0.3 for all formulations confirms that the systems are homogeneous. The potentials are decreased in their values, becoming more negative, and this could further help to prevent the aggregation of the nanoparticles. As regards the TPC, the values follow the order PNEG HA loa> PNEG PA loa> PNEG SG loa. These different values for the different phyto-nanoemulgels may likely be due to the different density values. Finally, the antioxidant activity, due to the polyphenol content of the GG extract, is preserved for all formulations even if there are some differences in their values that follow the order PNEG PA loa> PNEG HA loa> PNEG SG loa. In summary, as is conceivable, the order followed by the various polymers in relation to the AA% reflects the sequence of TPC values.

[0242] 3.4. Rheological Characterization of PNEG

[0243] 3.4.1. Viscosity as a Function of the Angular Deformation Rate of the PNEGs.

[0244] As can be seen in Figure 11, PNEGs show non-Newtonian behaviour with a shear-thinning effect in particular for the SG polymer. Loading the phyto-nanoemulgel with liquorice extract increases the viscosity for all formulations.

[0245] 3.4.2. A. Modulus G' and G "as a function of frequency for the loaded PNEG formulation.

[0246] Figure 12a shows that for the formulations of the polymers HA and SG the viscous modulus G" is prevalent. Elastic modulus G' tends to increase at higher frequencies. Similar to the loaded formulations, in Figure 12b, it can be seen that the viscous modulus G" is prevalent. The elastic modulus G' increases at higher frequencies, particularly for HA and PA polymers. In summary, the spectra are generally similar for PNEG HA and PNEG PA, with the viscous modulus always higher than the elastic modulus (G" > G') in the range of frequencies studied, highlighting a behaviour similar to that of a liquid. The elastic modulus is prevalent for PNEG SG as Solagum AX is composed of different types of polymers, such as gum arabic and xanthan gum, from which this different rheological characteristic derives, therefore the polymer influences the viscoelastic behaviour.

[0247] 3.4.3. Adhesion test for loaded and unloaded PNEGs

[0248] Table 16 shows the influence of different polymers on adhesiveness. In particular, the Peak Normal- Force / Normal Force (N) is related to stickiness and the Area under curve Force-Time (N-s) is a measure of adhesive force.

[0249] The results demonstrated that PNEG HA loa (loaded) has the highest value for tackiness and adhesive strength (-0.857 ±0.034N and 1.294±0.065 N.s, respectively). This could be explained by the chemical-physical characteristics of hyaluronic acid, a known gelling agent in cosmetic products. The formulations PNEG SG loa (-0.28410.015N and 0.693±0.045N.s), and PNEG PA loa (- 0.376±0.024N and 0.765±0.092N.s) have values lower than those of hyaluronic acid.

[0250] Table 16. Adhesion test for loaded and unloaded PNEGs

[0251] *Values are expressed as the average ± SD, (n = 6)

[0252] 3.4.5. Flow-curve of the loaded and unloaded PNEGs

[0253] The flow curves of the phyto-nanoemulgels are described in Figure 13 and Table 17. As can be seen from the profile of the curves, the shear-thinning effect and the pseudoplastic behaviour were confirmed for all formulations. Table 17. Flow-curve of loaded and unloaded PNEGs.

[0254] *Values are expressed as the average ± SD, (n = 3)

[0255] 3.4.6. Spreadability of loaded and unloaded PNEGs.

[0256] The measure of spreadability is a useful parameter in cosmetic formulation as it could favour the pleasantness of the product for users. As set forth in Table 18, PNEGs have similar spreadability values.

[0257] Table 18. Spreadability of loaded and unloaded PNEGs

[0258] *Values are expressed as the average ± SD, (n = 3)

[0259] 3.5. PNEG Stability Studies (Size, PDI, pH) All formulations were analysed for macroscopic appearance (visual inspection) after preparation and after 4 and 8 weeks of storage at room temperature (25 ± 2 °C), as shown in Table 19.

[0260] Table 19. Nanoparticle size, size distribution, and pH of formulations

[0261] Week O

[0262] Week 4

[0263] Week 8

[0264] 3.6. In vitro release studies

[0265] The in vitro release of TPC from phyto-nanoemulgels is shown in Figure 14. All formulations release the polyphenols more slowly than the extract solution GG. In particular, after 6 hours, the percentage of TPC released is 72.88% for the extract solution, while for PNEG HA loa, PNEG SG loa and PNEG PA Io it is 34.81±2.31, 50.14±0.05, 51.08±0.13%, respectively. The lower TPC value released by PNEG HA could be attributed to the higher viscosity of this formulation. Lowering the TPC released by phyto-nanoemulgels could improve topical absorption compared to DD extract solution.

[0266] 3.7. Physicochemical Characterization of Films Obtained from PNEG.

[0267] 3.7.1. Macroscopic appearance

[0268] Films from PNEGs were obtained following the procedure described in paragraph 2.6. After preparation, the films were removed from the Petri dishes and characterised prior to storage in the dryer. The films were observed macroscopically by visual and tactile inspection. The films obtained from PNEG PA showed good homogeneity , as shown in Figure 15. This film in particular has no rips and wrinkles and is relatively transparent, indicating that the vesicles are uniformly incorporated into the polymer matrix. Moreover, it is easy to remove and translucent.

[0269] 3.7.2. Determination of dry matter and thickness for PNEG PA loa films. The dry substance was determined by the difference between the weight of the empty Petri dishes and the weight after film formation. For PNEG PA loa it is 0.302±0.055 g while the film thickness is equal to 350 ± 0.04 pm.

[0270] 3.8. Cell viability studies

[0271] The biocompatibility of phytosomes loaded and unloaded with GG extract was tested by MTT assay on WS1 fibroblasts, i.e. connective tissue cells (Sal lustio et al., 2022). WS1 fibroblasts were treated for 24 h with the GG extract solution and loaded or unloaded phytosomes. The data (Figure 16) obtained demonstrate that the formulations are biocompatible and therefore safe for cells.

[0272] Further experiments, similar to those previously presented (Examples 1 and 2) were conducted on gels in which the ethosomal suspension contained extracts from fruits, leaves and roots of grapes, rosehip, centifolia rose, lavender, helichrysum, elderberry, pomegranate, yarrow, bergamot, mint, thyme, rosemary, oregano.

[0273] Bibliography

[0274] Brand-Williams, W., Cuvelier, M.E., Berset, C., 1995. Use of a free radical method to evaluate antioxidant activity. LWT - Food Sci. Technol. 28, 25-30. https: / / doi.org / 10.1016 / S0023- 6438(95)80008-5

[0275] Galassi, L., Rossi, M., Lodeserto, P., Lenzi, M., Borsetti, F., Voltattorni, M., Farruggia, G., Blasi, P., Orienti, I., 2023. Naxitamab Activity in Neuroblastoma Cells Is Enhanced by Nanofenretinide and Nanospermidine. Pharmaceutics 15, 648. https: / / doi.org / 10.3390 / pharmaceuticsl5020648

[0276] Rezvanian, M., Amin, M.C.I.M., Ng, S.-F., 2016. Development and physicochemical characterization of alginate composite film loaded with simvastatin as a potential wound dressing. Carbohydr. Polym. 137, 295-304. https: / / doi.Org / 10.1016 / j.carbpol.2015.10.091

[0277] Singleton, V.L., Orthofer, R., Lamuela-Raventos, R.M., 1999.

[0014] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent, in: Oxidants and Antioxidants Part A, Methods in Enzymology. Academic Press, pp. 152-178. https: / / doi.org / 10.1016 / S0076-6879(99)99017-l

[0278] Xie, J., Ji, Y., Xue, W., Ma, D., Hu, Y., 2018. Hyaluronic acid-containing ethosomes as a potential carrier for transdermal drug delivery. Colloids Surf. B Biointerfaces 172, 323-329. https: / / doi.Org / 10.1016 / j.colsurfb.2

Claims

CLAIMS1. A cosmetic solid composition comprising: a. phospholipid nanoparticles comprising a product obtained by extraction of a vegetal matrix with a hydroalcoholic mixture or with biosolvents; and b. a natural film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthan gum, pectin or mixtures thereof; and optionally c. a plasticizing agent, preferably glycerin; wherein said phospholipid nanoparticles are preferably ethosomes, phytosomes, liposomes and wherein said composition can be reconstituted into a fluid cosmetic by the addition of water at the time of application on the skin, wherein said composition is anhydrous, wherein said composition does not comprise preservatives,2. wherein said composition is in the form of a film. The cosmetic solid composition according to claim 1 wherein the phospholipid nanoparticles comprise soy lecithin.

3. The cosmetic solid composition according to any one of previous claims wherein said vegetal matrix is made of fruits or parts of plants, preferably said vegetable matrix is selected from blackthorn, grape, liquorice, rose hip, elderberry, pomegranate, yarrow, bergamot, mint, thyme, rosemary, oregano, centifolia rose, lavender, helichrysum.

4. The cosmetic solid composition according to any one of previous claims further comprising a vegetable oil, preferably selected from almond oil, olive oil, grapeseed oil, coconut oil, argan oil, jojoba oil, rose hip oil.

5. The cosmetic solid composition according to any one of previous claims wherein the film has a thickness of between 30 and 95 pm.

6. The cosmetic solid composition according to any one of previous claims reconstitutable in a fluid cosmetic in the form of gel, phytonanoemulgel, emulsion, lotion, serum.

7. A method for obtaining a solid cosmetic composition comprising the following steps: a. Extracting a vegetal matrix with a hydroalcoholic mixture or with biosolvents and evaporating the volatile portion to obtain a solid residue; b. Solubilizing the solid residue from step a. in water and treating it under stirring with a solution of phospholipid in water or ethanol to obtain a suspension of phytosomes or of ethosomes, preferably said phospholipid is soy lecithin; c. Adding to suspension obtained from step b. a film-forming polymer preferably selected from hyaluronic acid and its salts, sodium alginate, carrageenan, Arabic gum, xanthangum, pectin or mixtures thereof, and wherein said polymer is in a concentration comprised from 1% and 5% w / v, preferably 2% w / v; d. Optionally adding glycerin in concentration comprised from 1% and 3% w / v; e. Stirring and storing the mixture obtained from step d. for at least 12-24 hours to obtain the formation of a gel; f. Dehydrating the gel obtained from step e. to obtain a solid film.

8. The method according to claim 7 wherein the solution of soy lecithin in ethanol is added to the solution of solid residue in water at a rate comprised between 1 mL / min to 2 mL / min.

9. The method according to any one of claims 7 and 8 further comprising between the step b. and the step c. the addition of a vegetable oil preferably selected from almond oil, olive oil, grapeseed oil, coconut oil, argan oil, jojoba oil, rose hip oil and mixing to obtain a phyto- nanoemulsion.

10. The method according to any one of claims 7-9 wherein in step f. dehydration is carried out by a process selected from the group consisting of: drying, lyophilization.

11. The method according to claim 10 wherein dehydration is carried out by dehydration at a temperature comprised between 35 and 50°C, preferably 40 °C.

12. Cosmetic use of the composition according to any one of claims 1 to 6 or obtainable according to any one of claims 7 to 11 for skin care and cleansing, with a cosmetic action chosen from anti-aging, anti-dark spots, antioxidant, redensifying, protective, moisturizing, soothing, nourishing, anti-seborrheic, non-comedogenic, purifying action.

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