WO2021069698A1

WO2021069698A1 - Healing and conductive composition comprising collagen

Info

Publication number: WO2021069698A1
Application number: PCT/EP2020/078473
Authority: WO
Inventors: Emanuele MONDERNA
Original assignee: Monderna Health & Wellness
Priority date: 2019-10-09
Filing date: 2020-10-09
Publication date: 2021-04-15

Abstract

The invention relates to a composition suitable for topical application on the skin or mucous membranes, characterised in that it comprises at least water and at least a mixture of collagens, the collagen mixture comprising at least: - native collagen, the native collagen molecules having molecular weights ranging from 150 to 300,000 daltons, and - collagen gel, the collagen gel molecules having molecular weights ranging from 100 to 150,000 daltons. The invention also relates to such a composition and the use thereof in the treatment of skin and/or mucous membranes and a method for producing such a composition.

Description

HEALING AND CONDUCTIVE COMPOSITION INCLUDING

COLLAGEN

The present invention relates to a particular composition based on collagens for use on the skin and / or mucous membranes, in particular for healing and / or as a conductive cream for the treatment of the skin and / or mucous membranes by diathermy and / or radio frequency.

State of the art

Radiofrequency or diathermy is used in therapy and in cosmetics for various objectives such as, for example, to fight against the loss of skin tone, to fight against the redness of rosacea, to fight against cellulite, to promote the disappearance of residual marks on the skin. skin, etc.

Depending on the area to be treated and the desired goal, a metal return plate is positioned and the operator holds a handle in his hand. This handle can be ceramic (capacitive mode) or metal (resistive mode). According to a variant, the metal handle can be in polar, spherical or flat resistive mode, and in this case it is not necessary to use a return plate because the two polarizations are in the handle.

Between the manipulator and the subject's skin, the operator applies a neutral conductive cream without active principle. A magnetic field is then created thanks to the two polarizations. The body plays the role of conductor through its ions, which are charged water molecules. These molecules are mobilized and create endogenous heat. It results in an increase in the temperature of the tissues creating an activation of the dermis, which produces hyaluronic acid, collagen fibers and elastin fibers. This endogenous heat also induces vascularization, therefore an improvement in all internal exchanges. Thus the radiofrequency technique induces at the same time biochemical effects, mechanical effects and thermal effects.

Currently, radio frequencies oscillate between 300 KHz and 4 MHz, each frequency corresponding to a level of tissue resistance. The frequency used is determined by the area to be treated and the desired goal.

For the implementation of this technique and the proper functioning of the devices, it is mandatory to use a cream allowing the conduction of the waves between the manipulator and the tissue.

Current conductive creams are generally neutral and contain synthetic or petrochemical ingredients. The cream is conductive because it contains a majority of aqueous components. The existing conductive creams are not satisfactory because they are neutral creams without cosmetic or therapeutic efficacy as such, and therefore have no added value. Their only effect is to allow the conduction and the sliding of the manipulates. In addition, they are made up of petrochemical components, some with parabens and therefore unnatural and with endocrine disruptors. In addition, these creams have the property of fluffing, which makes it necessary to apply a larger quantity to allow the manipulation of the handle.

The objective of the invention is to overcome these various problems of the prior art.

Summary of the invention

According to the invention, the efficiency of the radiofrequency can be increased by introducing into the conductive cream molecules of collagen, molecules of natural origin, thus also allowing the use of this same cream without the application of radiofrequency.

This is why, to meet the objective of the invention, the inventor has developed a composition comprising a mixture of at least two kinds, two forms, of different collagens.

According to one variant, a composition comprising at least one particular collagen, namely at least one collagen obtained from sturgeon skins also makes it possible to meet the objective of the present invention.

Collagen is the most abundant structural protein in the body. By “collagen” within the meaning of the invention is meant either at least one particular form of collagen chosen from native collagen, gel collagen and hydrolyzed collagen, or a mixture of collagen comprising at least two collagen chosen from native collagen , a gel collagen and a hydrolyzed collagen, or a mixture of a native collagen, a gel collagen and a hydrolyzed collagen. It makes up about 70% of the connective tissue in the body and is necessary for building skin, muscles, bones and cartilage. Collagen ensures the cohesion, elasticity and regeneration of all these tissues. This protein is widely recognized as a functional ingredient thanks to its properties well known and identified for its health effects. 75% of the dermis is made up of collagen. From around 25-35 years of age, the body loses collagen at a rate of 1.5% per year.

Oral or topical collagen intake has been widely studied. Oral ingestion of collagen polypeptides has been shown to have positive effects on various body functions: bones, joints, skin, and tissues and ligaments.

The first uses of collagen in a beauty cream are quite old, and traces of its use can be found as early as 1936. The intensification of its use by the cosmetics industry is from the 1970s. This comes from studies carried out. on skin aging. Its role in the skin was known for a long time, but many teams then showed the modifications of the extracellular matrix during skin aging, and, in particular, the role and involvement of collagen in aging.

Collagen is characterized by its triple helix shape. There are traditionally three kinds or three forms of collagen:

Undenatured or native high molecular weight collagen. Native collagen has kept its native three-dimensional structure and the epitopes that allow it to interact with the immune system. Collagen in native form has not undergone any transformation and therefore corresponds to the entire protein. The native form is therefore difficult to assimilate by the organism. It is generally extracted gently at low temperature in an acidic medium because it is acid-soluble.

Collagen gel or collagen gel or medium molecular weight collagen gelatin. Collagen gelatin is generally obtained by hot aqueous extraction or chemical processes (strong acids or alkali) from skin, cartilage and other tissues containing collagen. Widely used for the food and pharmaceutical industry (capsule), it contains 84 to 90% of proteins and 1 to 2% of mineral salts. Hot extraction causes depolymerization of the collagen triple helix, it is denatured. A protein is thus obtained in gelled form with a more flexible format called collagen gel or collagen gel. Collagen gelatin thus differs from certain products on the market and called gelatin which are mainly products based on agar-agar, pectin and do not include collagen gelatin. By this transformation, the protein is more assimilated by the body than the native collagen.

Collagen hydrolyzate or low molecular weight hydrolyzed collagen. It is completely denatured collagen made up of all the amino acids that make up collagen. It is obtained by hydrolysis (enzymatic or acid) from raw materials containing collagen such as fish skins. It can also be obtained by hydrolysis of gel collagen. This hydrolysis splits the long collagen molecules into peptides and polypeptides (short chains of amino acids) which are bioavailable and therefore highly assimilable by the body.

There are also several sources of collagen. Bovine collagen is extracted from the skin of dead cows, sterilized and liquefied. It can be injected into the lips to swell them or under the wrinkles to fade them. But the bovine collagen ends up being absorbed by the body; this is the reason why its effects only last about three months. Marine collagen is also used in wrinkle creams. Marine collagen is extracted from residues from the food industry, most often fish or shrimp waste.

Collagen of marine origin is increasingly used and sought after as an alternative to collagen of mammalian origin, for religious and health questions (eg BSE crisis).

Marine collagen is used in cosmetics and cosmetics in many products with different properties highlighted, such as a moisturizing, humectant and / or a film-forming effect and / or an anti-aging, cellular restructuring effect.

However, collagen of marine origin, like collagen of bovine origin, is not entirely satisfactory and may contain heavy metals and residues which are not acceptable for use on the skin.

According to the invention, to meet the desired objectives, it is necessary to use at least gel collagen and / or native collagen and / or collagen from freshwater fish skins and in particular sturgeon skins. . Preferably, the species of sturgeon is Acipenser baerii and / or Acipenser gueldenstaedtii.

The collagens of sturgeon skins have a particular aminogram, since it contains about a third of Glycine and a high percentage of hydroxyproline and proline, and are of a higher purity than that of the collagens of marine fish.

Advantageously, the collagen obtained from the sturgeon skin makes it possible to avoid the presence of heavy metals in the collagens and therefore in the composition according to the invention. The sturgeons used are preferably reared in closed tanks supplied with excellent quality spring water and can receive a so-called organic diet. This helps to eliminate the entry of contaminants that are usually found in the skin of fish.

Thus, a subject of the invention is a composition suitable for topical application to the skin or the mucous membranes, comprising at least water and at least one mixture of collagens, said mixture of collagen comprising at least: native collagen, native collagen molecules having molecular weights between 150 and 300,000 Daltons, and gel collagen, the gel collagen molecules having molecular weights between 100 and 150,000 Daltons.

According to one variant, the invention relates to a composition suitable for topical application to the skin or the mucous membranes comprising at least one collagen obtained from sturgeon skins. Preferably, the composition according to the invention comprises at least water and at least said mixture of collagens, said collagen being obtained from sturgeon skins. Advantageously, such a composition in the form of a cream contains original components. natural, has good hold, has good conductivity, and can be used on the skin and / or mucous membranes for cosmetic or therapeutic applications, as a drug or medical device, alone or with the application of radiofrequencies

A subject of the invention is therefore also a composition for its use for the treatment of the skin and / or the mucous membranes, in particular as a medicament or a medical device, or alternatively as a cosmetic composition when it is applied to a skin and / or mucous membranes healthy (s) that is to say not diseased. In particular, the invention relates to the composition for its use in topical application to the skin and / or the mucous membranes for a healing effect or for its use in topical application to the skin and / or the mucous membranes as a conductive composition for the treatment of skin and / or mucous membranes by diathermy or radiofrequency. Brief Description of Figures

[Figure la]: Figure la is a photograph of a skin explant showing the general morphology of an injured area, without OJ treatment.

[Figure lb]: Figure lb is a photograph of a skin explant showing the general morphology of an injured area, without treatment on D4.

[Figure le]: Figure le is a photograph of a skin explant showing the general morphology of an injured area, with treatment with the composition according to the invention on D4.

[Figure 2a]: Figure 2a is a photograph of a skin explant exhibiting the general morphology without OJ treatment.

[Figure 2b]: Figure 2b is a photograph of a skin explant exhibiting the general morphology without treatment on D4.

[Figure 2c]: Figure 2c is a photograph of a skin explant exhibiting the general morphology with treatment with the composition according to the invention on D4.

[Figure 2d]: Figure 2d is a photograph of a skin explant showing the general morphology of an injured area, without treatment on D10.

[Figure 2e]: Figure 2e is a photograph of a skin explant exhibiting the general morphology with treatment with the composition according to the invention on D10.

[Figure 3a]: Figure 3a is a photograph of the abdomen of a 46 year old woman with postpartum stretch marks and scars following a tummy tuck.

[Figure 3b]: Figure 3b is a control photograph of the abdomen one month after the end of the treatment (3 radiofrequency sessions with application of a composition according to the invention - 3 sessions spread over 45 days).

[Figure 4]: Figure 4 is a graphical representation of the different forms of collagen (native, gel or hydrolyzed collagen); LMW meaning “Low Molecular Weight” that is to say low molecular weight; MMH meaning “Medium Molecular Weight”, that is to say medium molecular weight; HMW meaning "High Molecular Weight" that is to say high molecular weight.

Detailed description Composition according to the invention

A subject of the invention is therefore a composition suitable for topical application to the skin or the mucous membranes, comprising: at least water, and at least one mixture of collagens, said mixture of collagens comprising at least:

• native collagen, native collagen molecules with molecular weights between 150 and 300,000 Daltons, and

• gel collagen, the gel collagen molecules having molecular weights between 100 and 150,000 Daltons.

Preferably, the native collagen according to the invention has molecular weights of between 250,000 and 300,000 Daltons. Preferably, the gel collagen according to the invention has molecular weights of between 100,000 and 150,000 Daltons.

Another subject of the invention is a composition suitable for topical application to the skin or the mucous membranes, comprising at least one collagen obtained from sturgeon skin, preferably from sturgeon ventral skin. In particular, the sturgeon is I ’Acipenser baerii and / or I’ Acipenser gueldenstaedtii. In the present application, the singular and the plural are used interchangeably for the terms “skin” and “sturgeon”, the collagen (s) which may be derived from one or more skin (s) of one or more sturgeon (s). .

Preferably, the collagen (s) obtained from sturgeon skin comprises (comprise): a glycine content of between 21 and 26%, a hydroxyproline content of between 8 and 18%, a proline content of between 9 and 14%.

Preferably, the composition comprising at least one collagen obtained from sturgeon skin, also comprises at least water.

Preferably, the collagen (s) present in the composition according to the invention are type I collagens.

Preferably, the composition according to the invention comprises at least one mixture of collagens, preferably obtained from sturgeon skin, said mixture of collagens comprising at least :

• gel collagen, the gel collagen molecules having molecular weights between 100 and 150,000 Daltons, and possibly water.

According to one variant, the composition according to the invention comprises a mixture of at least two kinds or two forms of collagens, preferably obtained from sturgeon skin, chosen from: native collagen, native collagen molecules having weights molecular weight between 150 and 300,000 Daltons, preferably between 250,000 and 300,000 Daltons of collagen in gel, collagen molecules in gel having molecular weights between 100 and 150,000 Daltons, preferably between 100,000 and 150,000 Daltons, a collagen hydrolyzate, molecules of the collagen hydrolyzate having molecular weights between 500 and 5000 Daltons, and possibly water.

The mixture of collagens preferably comprises between 10 and 40% of native collagen and between 60 and 90% of collagen in gel, the percentages being given by weight relative to the total weight of dry matter of the mixture of collagens. According to one embodiment, the mixture of collagens comprises between 15 and 35% of native collagen and between 65 and 85% of collagen in gel, the percentages being given by weight relative to the total weight of dry matter of the mixture of collagens.

In the composition according to the invention, the collagen (s) obtained from sturgeon skin and / or the mixture of collagens comprising at least native collagen and gel collagen represents at least 0 , 1% by weight relative to the total weight of the composition, preferably between 0.1 and 10%, more preferably between 0.1 and 5%.

According to the invention, it is preferred for the native collagen to be obtained by carrying out a process at a temperature below 35 ° C., so as to preserve the entire structure of the collagen without denaturing.

The mixture of collagens used in the compositions according to the invention can be of any origin. They can be, for example, collagens of mammalian origin, such as bovine collagens. extracts from the skin of dead cows.

It can also be marine collagens. Marine collagen can be extracted from food industry residues, such as fish or shrimp waste, or directly from fish skins. Advantageously, marine collagen has a greater absorption capacity than collagen of mammalian origin. Furthermore, marine collagen has a higher average proline and hydroxyproline content than animal collagen, which gives it less mechanical strength and lower viscosity.

It can also be collagens obtained from the skins of freshwater fish.

According to a preferred embodiment, the composition according to the invention comprises collagens obtained from fish skins. Preferably, the collagens are obtained from freshwater fish skins and even more preferably from sturgeon skins, and in particular from sturgeon ventral skins.

Advantageously, the collagen obtained from the skin of sturgeons makes it possible to avoid the presence of heavy metals in the collagens and therefore in the composition according to the invention. The sturgeons used are preferably reared in closed tanks supplied with excellent quality spring water and can receive a so-called organic diet. This helps to eliminate the entry of contaminants that are usually found in the skin of fish.

According to a particularly suitable embodiment, the collagen (s) used in the compositions according to the invention exhibit an aminogram exhibiting at least the following characteristics: a glycine content of between 21 and 26%, a content in hydroxyproline between 8 and 18%, a proline content between 9 and 14%.

In addition to the mixture of collagens and / or of the sturgeon skin collagen (s), the composition according to the invention comprises at least water. Preferably, the composition according to the invention comprises between 50 and 95% water by weight relative to the total weight of the composition, very preferably between 65 and 80% water.

The water present in the composition according to the invention can be of any origin.

Preferably at least part of the water present in the composition according to the invention is source water. Preferably at least 3% of the water present in the composition according to the invention is spring water, the percentage being given by weight relative to the total weight of water present in the composition.

Preferably at least part of the water present in the composition according to the invention is water of marine origin.

According to one embodiment, at least part of the water present in the composition is water from a marine source. Preferably at least 3% of the water present in the composition according to the invention is marine spring water, the percentage being given by weight relative to the total weight of water present in the composition.

In addition to the water and the mixture of collagens, the composition according to the invention can comprise other constituents, these having to be dermatologically and / or cosmetically acceptable. They may in particular be excipients or active ingredients. Preferably, the composition according to the invention comprises: at least one vegetable oil, such as for example a sunflower or pomegranate or grape seed oil, and / or at least one active principle which heals and / or moisturizes and / or antibacterial and / or soothing and / or anti-irritant, such as for example squalane or aloe vera gel, and / or at least one plant extract, such as for example extract of punica granatum, or extract of flower of sophora japonica, or leaf extract rosemary, and / or at least one emulsifier, such as for example Sorbitan isostearate or polyacrylamide or polysorbate 20, and / or at least one emollient, such as for example octyldodecanol or propanediol or caprylic, and / or at least one preservative, such as for example potassium sorbate, sodium benzoate, benzyl alcohol, dehydroacetic acid, and / or at least one stabilizer, such as for example tocopherol, and / or glycerin.

The composition according to the invention can be provided in various forms suitable for topical application to the skin and / or the mucous membranes. It may in particular be a cream, a serum or a gel or a solution.

Process for manufacturing the composition according to the invention

The composition according to the invention can be obtained by any suitable manufacturing process.

In particular, the composition according to the invention can be obtained by a manufacturing process comprising at least the implementation of the following steps: obtaining at least one solution of collagen (s) from sturgeon skins, in the case of where there are several solutions of collagen (s), mixture of solutions of collagen (s), mixture of solution obtained with the possible other constituents of the composition. The composition according to the invention can also be obtained by a manufacturing process comprising at least the implementation of the following steps: a. obtaining a native collagen solution, preferably from sturgeon skins, b. obtaining a gel collagen solution, preferably from sturgeon skins c. mixing the two solutions to obtain a solution of native collagen and gel collagen, d. mixing of the constituents of the composition at room temperature.

The native collagen solution can be obtained anywhere suitable medium.

According to one embodiment, step a. includes the implementation of the following steps:

Washing of the raw material containing collagen, preferably fish skins, even more preferably sturgeon skins and in particular sturgeon ventral skins; the washing is preferably carried out in two sub-steps: o a basic washing with sodium hydroxide NaOH, to clean the skins o then with water to neutralize extraction with acid, filtration and centrifugation, precipitation of the collagen by NaCl and centrifugation, solubilization by addition of acid, dialysis to remove the salt (NaCl), tangential concentration.

The process for obtaining native collagen is preferably carried out at a temperature below 35 ° C. It thus keeps its triple helix and its native collagen properties.

The gel collagen solution can be obtained by any suitable means. According to one embodiment, step b. includes the implementation of the following steps:

Washing of the raw material containing collagen, preferably fish skins, even more preferably sturgeon skins and in particular sturgeon ventral skins; the washing is preferably carried out in two sub-steps: o a basic washing with sodium hydroxide NaOH, to clean the skins o then with water to neutralize extraction with acid filtration and centrifugation precipitation of collagen by NaCl and centrifugation, solubilization by addition of dialysis acid to remove the salt (NaCl) tangential concentration, cooking at 60 ° C., preferably with stirring, sieving, filtration through activated carbon.

The two solutions (native collagen and gel collagen) are then mixed to form a collagen solution.

The collagen solution is then mixed with water and with the other optional constituents of the composition, preferably at room temperature in order to preserve the original qualities of the active agents and in particular of the ingredients of natural origin constituting the composition according to the invention. Advantageously, the implementation of the process at ambient temperature is of economic and environmental interest.

Uses of the composition

The composition according to the invention has an effectiveness on the skin and / or the mucous membranes, in particular a healing effectiveness.

A subject of the invention is therefore also the composition for its use in the treatment of the skin and / or the mucous membranes. The composition can be used as a medicament, essentially as a dermatological medicament, or as a medical device or else as a cosmetic or dermocosmetic composition.

In particular, a subject of the invention is the composition for its use in topical application to the skin and / or the mucous membranes for a healing effect.

The invention also relates to the use of the composition on healthy skin or mucous membranes in topical application for a cosmetic effect, in particular for an anti-aging cosmetic effect, for preventing and / or combating skin aging.

Thus, the invention is also aimed at cosmetic use on healthy skin or mucous membranes in topical application for an anti-aging cosmetic effect, to prevent and / or fight against normal aging of the skin.

Advantageously, the composition according to the invention can also be used as a conductive cream for the treatment of the skin and / or the mucous membranes by diathermy and / or radiofrequency. The therapeutic or cosmetic use of the composition according to the invention can therefore consist of this. A subject of the invention is also the composition for its use. in topical application to the skin and / or the mucous membranes as a conductive composition for the treatment of the skin and / or the mucous membranes by diathermy and / or radiofrequency.

Also the composition according to the invention can be used with or without application of radiofrequencies. In application with radiofrequencies, the composition is preferably applied in a circular manner to the area to be treated with the radiofrequency device so as to cover the entire surface and approximately 2 mm in thickness. The practitioner then passes the manipulation over the area to be treated by following a protocol adapted to each pathological or cosmetic problem to be treated. If necessary, it is possible to add more cream while the protocol is being carried out. The conductivity of the composition according to the invention is high.

Advantageously, at the end of the session, it is not necessary to wipe off the composition as is the case with current conductive creams or conductive gels. Indeed, thanks to its dual use, the composition according to the invention is absorbed by the skin and acts as such through its intrinsic properties, thus supporting the recognized cosmetic or therapeutic regenerating properties of radiofrequency.

The radio frequency causes vasodilation in the area of application, increasing blood flow and raising the temperature locally causing better supply of nutrients and oxygen, and the elimination of catabolites. The effects of radiofrequency temperature cause the denaturation of collagen in the dermis, promoting immediate and effective contraction of its fibers, activating fibroblasts and leading to neo-collagenization leading to the reorganization of collagen fibers and tissue remodeling. During the treatment, the temperature on the surface of the skin reaches 40 to 42 ° C, and 50 to 60 ° C in depth. The native collagen present in the composition according to the invention is denatured during the use of radiofrequency. The triple helix chain is undone under the effect of heat (from 35 ° C); it penetrates the dermis more easily because its molecular weight will be lower and the vasodilation of the pores promotes its penetration into the dermis. This reinforces the effect of the collagen gel. Radio frequency can take collagen molecules deeper into the layers of the dermis.

The invention is now illustrated by examples and test results.

EXAMPLES

Example of composition according to the invention

An example of a composition according to the invention, which is in the form of a cream, is obtained by mixing constituents at room temperature, which in the finished composition consists of (the contents being given in percentage by weight relative to the weight of the composition ): native collagen of sturgeon skins: 0.01 to 1 collagen gel of sturgeon skins: 0.01 to 1 glycerin: 0.2 to B squalane: 3 to 6 pomegranate oil: 1 to 2 flower extract of sophora japonica: 0.001 to 0.05 pomegranate extract: 0.001 to 0.05 essential oil of niaouli: 0.01 to 0.1 sunflower oil: 0.1 to 1 marine spring water: 2 to 5 - Water: 75 to 88 caprylic / capric triglyceride: 5 to 8 emulsifier: 1 to 3 preservatives: QS

Demonstration of the effectiveness of the composition according to the invention

I. Study on skin explants of the healing and anti-aging effect of the composition according to the invention Various products were tested containing different concentrations of collagen and different forms of collagen.

The different compositions tested are described in Table 1 (collagen content values, percentage by weight relative to the total weight of the composition; the remainder of the composition is that of the example of composition according to the invention).

[Table 1]

The healing and anti-aging effect tests of these compositions were carried out on skin explants. For the two tests, the results were obtained after histological treatment and study of the general morphology:

Histological treatments: After 24 hours in buffered formalin, the samples were dehydrated and impregnated with paraffin using a Leica PEARL automatic dehydration machine. they have was en bloc using a Leica EG 1160 embedding station. 5 µm sections were made using a Minot type microtome, Leica RM 2125 and mounted on Superfrost histological glass slides. ^® . The frozen samples were cut into sections 7 μm thick using a Leica CM 3050 cryostat. The sections were then mounted on Superfrost ^® Plus glass slides. The microscopic observations were carried out by optical microscopy, using a Leica DMLB or Olympus BX43 type microscope. The shots were taken with an Olympus DP72 camera and Cell ^A D software.

General morphology: The general morphology was observed on paraffin sections after staining with Masson's trichrome variant of Goldner. It was assessed by microscopic examination.

1.1. Healing effect of the composition according to the invention

In order to test the healing properties of the composition according to the invention (P3) and of the other products P1 to P5, a study was carried out on explants of skin.

The operating protocol for the test is as follows:

Preparation of explants: 6 rectangular explants of 10x15 mm, with wound per batch, were prepared from an abdominoplasty of a 60-year-old Caucasian woman. The explants were left to survive in a culture medium at 37 ° C. in a humid atmosphere, enriched with 5% CO 2.

Creation of lesions by mechanical injury: On D0 on the explants, two wounds by explant were performed using a 3.5 mm diameter biopsy punch. The lesions were produced just before the application of the tested products.

Application of the products: On D0 (after injuries) and D3, D5 and D7, the tested products were applied topically, at a rate of 3 μl for the rectangular explants in order to obtain a concentration of 2 mg per cm ² . The products were successively spread using a spatula.

The explants of the control batches received no treatment except for the renewal of the culture medium, half (1mL / well) on D3.

Samples: On D0, the 3 explants of batches T0 and Bo were taken and cut in half. One part was fixed in buffered formalin solution, and the other was frozen at -80 ° C. On D4 and D10, 3 explants from each batch were taken and treated in the same way as on D10.

The results obtained with composition P3 are shown in Figures la (general morphology), lb (control at D4) and (invention at D4).

Figure la shows the general morphology of the injured area. There is no growth bud.

FIG. 1b represents the general morphology of the injured area of the untreated control on D4. The growth bud has a length of 150 µm.

Figure 1c shows the general morphology of the injured area after 4 days of treatment with the composition according to the invention (PB). The general morphology is good in the epidermis and in the dermis. The stratum corneum has a thin and slightly laminated appearance, the relief of the dermo-epidermal junction (DEJ) is low and the density of the papillary dermis is clear.

It is observed that the composition according to the invention, after 4 days of treatment, exhibits greater healing activity than the control, mainly by promoting both the proliferation and the migration of the keratinocytes of the growth bud.

In addition, the results obtained for the products PI to P5 are presented below:

The PI product, after 4 days of treatment, exhibits healing activity mainly by promoting the migration of keratinocytes and therefore the advancement of the growth bud. This activity is no longer observed after 10 days.

The product P2 after 4 days of treatment exhibits healing activity, mainly by promoting the proliferation of keratinocytes in the growth bud.

The product P3, after 4 days of treatment, exhibits healing activity, mainly by promoting both the proliferation and the migration of keratinocytes from the growth bud. After 10 days of treatment, it only promotes the proliferation of keratinocytes in the growth bud.

The product P4, after 4 days of treatment, does not exhibit any healing activity. But after 10 days of treatment, it promotes the proliferation of keratinocytes in the growth bud.

The product P5, after 4 or 10 days of treatment, does not exhibit any healing activity.

Thus the association of native collagen and gel collagen is important to obtain the healing effect.

1.2. Anti-aging effect of the composition according to the invention

In order to test the anti-aging properties of the composition according to the invention (P3) and of the other products P1 to P5, a study was carried out on skin explants.

The operating protocol for the test is as follows:

- Preparation of explants: 45 circulars with a diameter of 12 ± 1 mm, were prepared from an abdominoplasty of a 60-year-old Caucasian woman (reference: P2124-AB60). The explants were left to survive in culture medium at 37 ° C in a humid atmosphere, enriched with 5% C02.

- Application of the products: On D0, D3, D5 and D7, the products tested were applied topically, at a rate of 2 mg per cm ² . The products were successively spread using a spatula.

The explants of the control batches received no treatment except for the renewal of the medium. of culture, for half (lmL / well) on JB, D5 and D7.

Samples: At OJ, the 3 explants of the T0 batches were taken and cut in two. One part was fixed in buffered formalin solution, and the other was frozen at -80 ° C. On D4 and D10, 3 explants from each batch were taken and treated in the same way as in OJ. According to the provisions provided for in the study plan, the days of treatments and samples were modified to adapt the planning of the study according to the working days.

The results are presented in FIGS. 2a (general morphology), 2b (control on D4), 2c (invention on D4), 2d (control on D10) and 2e (invention on D10).

FIG. 2a represents the morphology of the explant on normal skin. The general morphology is good in the epidermis and in the dermis. The stratum corneum has a thin and slightly laminated appearance, the relief of the dermo-epidermal junction (DEJ) is low and the density of the papillary dermis is clear.

FIG. 2b represents the morphology of the explant without treatment on D4: The general morphology is quite good in the epidermis and good in the dermis. The stratum corneum presents a moderately thick and laminated appearance, the relief of the dermo-epidermal junction (DEJ) is moderate and the density of the papillary dermis is moderate.

FIG. 2c represents the morphology of the explant after 4 days of application of the composition P3: the invention induces an increase in the density of the papillary dermis.

FIG. 2d represents the morphology of the explant without treatment after 10 days: the general morphology is very slightly altered in the epidermis and good in the dermis. The stratum corneum presents a rather thick and moderately laminated appearance, the relief of the dermo-epidermal junction (DEJ) is moderate and the density of the papillary dermis is low.

FIG. 2e represents the morphology of the explant after 10 days of application of the composition P3: the invention induces an increase in the density of the papillary dermis.

It is observed that the composition according to the invention is well tolerated by the skin and does not induce any deterioration in cell viability in the epidermis or in the dermis. On D4, the composition according to the invention exhibits anti-aging activity by improving the density of the dermis which is clearly visible, and which continues on D10.

- The PI product, after 4 days of treatment, does not induce an increase in the density of the papillary dermis. An increase is seen after 10 days.

- The product P2, after 4 days of treatment, does not induce an increase in the papillary dermis. - The PB product, after 4 days of treatment and after 10 days of treatment, induces an increase in the density of the papillary dermis.

- The product P4, after 4 days of treatment, does not induce an increase in the density of the papillary dermis. An increase is seen after 10 days.

- The product P5, after 4 days of treatment does not induce an increase in the density of the papillary dermis.

II. Effect of the composition according to the invention with the application of radiofrequencies

Tests carried out with the application of radiofrequencies show significant conductivity.

In addition, an example of efficiency is shown below.

- Subject: 46-year-old woman with 15-year-old postpartum stretch marks and the scar from a 2.5-year-old tummy tuck.

- Equipment used: Fisioline brand radiofrequency device, Radiant model

- Protocol used: Preparation of the patient (installation of the return plate). Application of the composition of the example to the treated area then passage of the manipulator in circular motion for 7 to 15 min according to a specific program:

- ^1st switch to multifrequency in capacitive mode

- ^2nd passage in resistive mode

- last switch to bipolar mode.

Two other sessions take place following the same process at 15 day (2 week) intervals, ie 3 sessions over a period of treatment carried out within 45 days.

The results are shown in Figures 3a and 3b.

Figure 3a is a photograph of the abdomen of a 46 year old woman with postpartum stretch marks and scarring following a tummy tuck. We see marked stretch marks with a "scale skin" appearance, and adherent vertical and horizontal scars.

FIG. 3b is a control photograph of the abdomen one month after the end of the treatment (3 radiofrequency sessions with application of the composition according to the invention spread over 45 days). The skin is smoother, toned and elastic. The horizontal and vertical scar is reduced. Stretch marks are extremely reduced. The adhesion of the vertical scar has disappeared.

Claims

[Claim 1] Composition suitable for topical application to the skin or the mucous membranes, characterized in that it comprises at least water and at least one mixture of collagens, said mixture of collagens comprising at least:

- native collagen, native collagen molecules having molecular weights between 150 and 300,000 Daltons, and

- gel collagen, the gel collagen molecules having molecular weights between 100 and 150,000 Daltons.

[Claim 2] Composition according to the preceding claim, characterized in that the collagen is obtained from sturgeon skin.

[Claim 3] Composition according to one of the preceding claims, characterized in that the collagen (s) comprises (comprise):

- a Glycine content of between 21 and 26%,

- a hydroxyproline content of between 8 and 18%,

- a proline content of between 9 and 14%.

[Claim 4] Composition according to one of the preceding claims, characterized in that it comprises at least 0.1% of collagens by weight relative to the total weight of the composition.

[Claim 5] Composition according to one of the preceding claims, characterized in that it comprises between 0.1 and 10% of collagens by weight relative to the total weight of the composition.

[Claim 6] Composition according to one of the preceding claims, characterized in that it comprises between 50 and 95% water by weight relative to the total weight of the composition.

[Claim 7] Composition according to one of the preceding claims, characterized in that the mixture of collagens comprises between 10 and 40% of native collagen and between 60 and 90% of collagen in gel, the percentages being given by weight relative to the total weight of dry matter of the collagen mixture.

[Claim 8] Composition according to one of the preceding claims, characterized in that the native collagen has been obtained by a process carried out at a temperature below 35 ° C.

[Claim 9] Composition according to one of the preceding claims, characterized in that at least part of the water is spring water.

[Claim 10] Composition according to one of the preceding claims, characterized in that at least part of the water is of marine origin.

[Claim 11] Composition according to one of the preceding claims, characterized in that it also comprises:

- at least one vegetable oil, and / or

- at least one healing and / or moisturizing and / or antibacterial and / or soothing and / or anti-irritation active principle,

- at least one plant extract, and / or

- at least one emulsifier, and / or

- at least one emollient, and / or

- at least one preservative, and / or

- at least one stabilizer, and / or

- glycerin.

[Claim 12] Composition according to one of the preceding claims, characterized in that it is provided in the form of a cream, a serum, a solution or a gel.

[Claim 13] Composition according to one of the preceding claims, for its use in the treatment of the skin and / or mucous membranes.

[Claim 14] Composition according to one of claims 1 to 12, for its use in topical application to the skin and / or the mucous membranes for a healing effect.

[Claim 15] Composition according to one of claims 1 to 12, for its use in topical application to the skin and / or the mucous membranes as a conductive composition for the treatment of the skin and / or the mucous membranes by diathermy and / or radiofrequency.

[Claim 16] Composition according to one of Claims 1 to 12, for its cosmetic use in topical application to healthy skin and / or to healthy mucous membranes.

[Claim 17] A composition for its use according to the preceding claim, for preventing and / or combating skin aging.

[Claim 18] A method of manufacturing a composition according to one of claims 1 to 12, characterized in that it comprises at least the implementation of the following steps: a. obtaining a solution of native collagen, b. obtaining a gel collagen solution, vs. mixing the two solutions to obtain a mixture of native collagen and gel collagen, d. mixing of the constituents of the composition at room temperature.