The present invention relates to the use of neutral core human milk oligosaccharides such as in particular lacto-N-(neo)tetraoses as anti-ageing active ingredient as well as the use thereof in topical anti-ageing compositions.

The human skin is the biggest organ of the human body, and it is the barrier that separates the body from the outer environment. Human skin is made up of several layers that are classified as epidermis, dermis and subcutaneous tissue. The epidermis is the outermost layer of the skin and is mainly formed of keratinocytes comprising about 90% of the epidermis and melanocytes which are found at the base of the epidermis and make melanin. The main function of epidermis is to form a permeability barrier against environmental challenges, such as UV radiation, heat, chemicals, pollution, and pathogens, such as bacteria, fungi, parasites, and viruses. It also protects the body from uncontrolled water evaporation from inside out, maintaining the hydration balance and skin metabolism.

The dermis is the second layer of skin. In the dermis, the most abundant cell type are the dermal fibroblasts, which are responsible of generating the connective tissue by producing extracellular matrix (ECM). Said ECM is a three-dimensional network consisting of extracellular macromolecules and minerals, such as collagen, enzymes, glycoproteins and hydroxyapatite. The ECM imparts strength and integrity to the skin.

As a major structural protein of the extracellular matrix (ECM), type I collagen is secreted by fibroblasts, forming more than 90% of the dry weight of the dermis. It is produced as procollagen which is then processed to mature collagen.

The aging process is a dynamic and unchangeable phenomenon which affects all systems in the body. It is known that endogenous (age-related) or exogenous (chronic light exposure, heat, pollution, chemicals, toxins) aging leads to an irreversible degeneration of tissues, in particular of skin. In skin, visible signs of ageing are inter alia fine-lines, wrinkles and sagging. Modifications in collagen, the most important protein of the connective tissue, are the main factors for these anatomic changes. Even though collagen is constantly produced and recycled, with increasing age the rate of synthesis of collagen significantly reduces thus leading to the visible manifestations of skin ageing.

Thus, there is an ongoing need for preferably natural ingredients which are able to increase collagen production and are thus able to counteract skin-ageing and the visible signs associated therewith.

Surprisingly it has now been found that certain human milk oligosaccharides namely the neutral core human milk oligosaccharides composed of galactose, N-acetylglucosamine, and glucose monosaccharide such as Lacto-N-Tetraose (LNT) or Lacto-N-Neotetraose (LNnT) are able to significantly increase collagen synthesis in human dermal fibroblasts.

Thus, in a first embodiment, the present invention concerns a non-therapeutic use of a neutral core human milk oligosaccharide composed of galactose, N-acetylglucosamine, and glucose monosaccharide for preventing, reducing or treating skin aging in a subject (in need thereof).

In another aspect, the invention concerns neutral core human milk oligosaccharide composed of galactose, N-acetylglucosamine, and glucose monosaccharide for use in prevention, reduction or treatment of skin aging in a subject.

In a further aspect, the invention concerns a method of preventing, reducing or treating skin aging in a subject, said method comprising administering a neutral core human milk oligosaccharide composed of galactose, N-acetylglucosamine, and glucose monosaccharide to said subject, in particular to a skin area in need thereof such as e.g. to facial skin, skin of the hands and photoaged skin and optionally appreciating the effect.

Collagen is responsible for skin strength and structure; collagen homeostasis in the deeper part of the skin is maintained by balancing the degradative and synthetic pathways and a skewed balance between collagen production and degradation can lead to skin aging. It has been found in accordance with the present invention, that the core human milk oligosaccharides composed of galactose, N-acetylglucosamine, and glucose monosaccharide increase type I collagen in human dermal fibroblasts and are thus able to maintain skin homeostasis, in particular in ageing and/or aged skin.

Thus, the present invention also relates to the use of neutral core human milk oligosaccharides composed of galactose, N-acetylglucosamine, and glucose for inreasing (type I) collagen production in fibroblasts respectively in the skin (compared to an untreated control).

The invention also relates to a use of a neutral core human milk oligosaccharide composed of galactose, N-acetylglucosamine, and glucose monosaccharide for increasing the expression of (type I) collagen in fibroblasts respectively in the skin (compared to an untreated control).

The invention also relates to a non-therapeutic method for stimulating collagen I synthesis in skin cells, preferably fibroblasts by topically applying a neutral core human milk oligosaccharide composed of galactose, N-acetylglucosamine, and glucose monosaccharide or a composition comprising said a neutral core human milk oligosaccharide as defined herein to a skin area in need thereof.

Preferably, in all embodiments of the present invention, the fibroblasts are human fibroblasts, most preferably dermal human fibroblasts.

The term ‘neutral core human milk oligosaccharides’ (neutral core HMOs) as used herein refers to a family of structurally diverse unconjugated glycans that are highly abundant in and unique to human milk and which are built up from galactose, N-acetylglucosamine and glucose monosaccharide units, preferably linked through 0-(1 -3) or 0-(1 -4) bonds with each other. Preferably, in all embodiments herein, the galactose is D-galactose and the glucose Is D-glucose.

In all embodiments of the present invention, preferably, the neutral core human milk oligosaccharides are tri-, tetra-, hexa-, octa- or decasaccharides, with the tri-, tetra- and hexasaccharides being particularly preferred. Most preferred are the tetrasaccharides. In all embodiments of the present invention, it is further advantageous that the oligosaccharides are linear oligosaccharides, i.e. not branched.

Exemplary oligosaccharide according to the present invention are depicted below

It is furthermore advantageous in all embodiments of the present invention, that the oligosaccharides are characterized by a terminal D-galactose linked through a (3-(1 - 3) or a (3-(1 -4) bond to a N-acetyl-d-glucosamine (GIcNAc) unit.

In addition, in all embodiments of the present invention, it is also preferred that the oligosaccharides are characterized by D-galactose linked through a (3-(1 -4) bond to a reducing end D-glucose.

Particularly suitable neutral core human oligosaccharides according to the present invention are listed in table 1

Advantageously, in all embodiments of the present invention lacto-N-triose II, LNT, LNnT, LNH, LNnH, pLNH and pLNnH as well as mixtures thereof are used. Most preferred in all embodiments of the present invention is the use of Lacto-N- (neo)tetraoses, i.e. lacto-N-tetraose (CAS No: 14116-68-8) and/ or lacto-N- neotetraose (CAS No: 13007-32-4). LNnT is the most preferred as it is most active in inducing collagen synthesis. HMOs can be isolated from breast milk or they can be produced chemically or biochemically according to methods well known in the art.

For the purpose of the present invention the source of the HMO is not essential. It is clear that HMOs from different sources can be used. The term "subject", as used herein, means an animal having a skin that separates and defines the animal inside from the outside. Preferably, the subject is a human.

In one embodiment of the present invention the subject may be female. In another embodiment the subject may be male. In a further embodiment the subject may be with a non-binary gender.

In a preferred embodiment the subject is not a child. The term ‘child’ as used herein means a human Io 7 years of age or younger. Thus, in all embodiments of the present invention, the subject is a human that is 8 years of age or older, preferably the subject is a human being 20 years or older, preferably 30 years or older, even more preferably 40 years or older such as most preferably 50 years or older.

In a further embodiment, the prevention, reduction or treatment of the skin aging in a subject is attained by increasing or maintaining the production of type I collagen or slowing down the reduction of type I collagen in the skin and/or maintaining collagen metabolism or slowing down the reduction of the collagen metabolism in the skin.

Preferably, in all embodiments of the present invention the collagen I expression (relative to untreated control) is increased by at least 10%, more preferably by at least 20%, most preferably by at least 30%, such as by at least 50 %, most preferably in normal human dermal fibroblasts.

In a further embodiment, the present invention relates to the prevention, reduction or treatment of the skin aging caused by intrinsic and/ or extrinsic factors, such as, but not limited to sun, sunburn, collagen damage, senescence, infrared radiation, heat, hormonal reasons, malnutrition, temperature, tobacco smoking, stress, sleep deprivation, pollution, or alcohol consumption.

The term ‘prevention, reducing or treating skin aging’ includes the prevention, reduction or treatment of fine lines, wrinkles, crow’s feet, sagging, skin thinning and/ or rough skin texture as well as improving skin elasticity or skin firmness without being limited thereto.

The term ‘prevention, reducing or treating skin aging’ also encompass smoothening of wrinkles and fine lines as well as decreasing their volume and depth in a person in need thereof. In all embodiments of the present invention preferably the use is non-therapeutic, i.e. a cosmetic use intended for beautifying the skin.

In all embodiments of the present invention, the neutral core HMOs are applied to an external surface of the human body.

The term ‘external surface of the human body’ as used herein encompasses the skin as well as the scalp. Preferably, in all embodiments of the present invention the external surface of the human body treated according to the present invention is the face, neck and/or body skin, most preferably the face (including the (lateral) cheek, forehead, nose, chin).

In all embodiments of the present invention, the neutral core HMOs are preferably administered in the form of a cosmetic or dermatological composition.

The term ‘cosmetic or dermatological composition’ as used herein refers to cosmetic or dermatological compositions which are used to treat, care for or improve the appearance of the skin and/or the scalp, preferably the skin, most preferably facial skin and/ or the skin of the hands, i.e. which are intended to beautify the skin and/ or scalp.

The term ‘dermatological composition’ as used herein refers to compositions which are used to be applied (topically) to skin.

The total amount of the neutral core HMO(s) according to the present invention in the compositions according to the present invention is preferably selected in the range from 0.01 to 10 wt.-%, more preferably in the range from 0.1 to 7.5 wt.-%, most preferably in the range from 0.2 to 5 wt.-%, based on the total weight of the composition. Further suitable ranges are from 0.25 to 2.5 wt.-%, from 0.5 to 2 wt.-%, from 0.1 to 1 wt.-%, from 0.25 to 0.75 wt.-% and from 0.3 to 0.6 wt.-%. Particularly preferred ranges according to the present invention are from 0.1 to 5 wt.-%, more preferably from 0.25 to 5 wt.-%, such as from 0.3 to 5 wt.-%.

If nothing else is stated in this specification any given parts and percentages are per weight and based on the total weight of the composition. In a particular embodiment, the compositions according to the present invention are cosmetic or dermatological compositions intended to be topically applied to mammalian keratinous tissue such as in particular to human skin or the human scalp. Such compositions are also called dermatological compositions. Thus, preferably in all embodiments of the present invention the cosmetic or dermatological compositions are topical cosmetic (i.e. dermatological) compositions with all the definitions and preferences as given herein.

It is well understood that said compositions may comprise only one or several HMOs as defined herein.

The topical cosmetic or dermatological compositions according to the present invention may be leave-on or rinse-off compositions, and include any product applied to a human body, primarily for improving appearance or general aesthetics. Preferably the cosmetic or dermatological compositions of the present invention are leave-on compositions.

It is well understood that the cosmetic or dermatological compositions according to the invention intended for topical application comprise a physiologically acceptable medium, i.e. a medium compatible with keratinous substances, such as the skin, mucous membranes, and keratinous fibers. In particular, the physiologically acceptable medium is a cosmetically acceptable carrier. In all embodiments of the present invention, it is preferred that the carrier comprises water.

The term ‘cosmetically acceptable carrier’ (also referred to herein as carrier) refers to all vehicles/ carriers conventionally used in cosmetic or dermatological compositions, i.e. which are suitable for topical application to the keratinous tissue, have good aesthetic properties, are compatible with the actives present in the composition, and will not cause any unreasonable safety or toxicity concerns. Such carriers are well- known to one of ordinary skill in the art and can include one or more compatible liquid(s) or solid filler diluent(s), excipient(s), additive(s) or vehicle(s) which are suitable for application to skin.

The exact amount of carrier will depend upon the actual level of the active ingredients and of any other optional ingredients that one of ordinary skill in the art would classify as distinct from the carrier (e.g., other active ingredients). In an advantageous embodiment, the cosmetic or dermatological compositions according to the present invention comprise from about 50% to about 99%, preferably from about 60% to about 98%, more preferably from about 70% to about 98%, such as in particular from about 80% to about 95% of a carrier, based on the total weight of the cosmetic or dermatological composition.

In a particular advantageous embodiment, the carrier in the the cosmetic or dermatological compositions according to the present invention consists of at least 30 wt. %, more preferably of at least 40 wt.-%, most preferably of at least 45 wt.-% of water, such as in particular of 50 to 90 wt.-% of water.

The cosmetic or dermatological compositions in accordance with the invention can be in the form of a liquid, lotion, a thickened lotion, a gel, a cream, a milk, an ointment, a paste, a powder, a make-up, or a solid tube stick and can optionally be packaged as an aerosol and can be provided in the form of a mousse such as a aerosol mousse, a foam or a spray foam, a spray, a stick.

Preferably the one or more neutral core HMOs according to the present invention are formulated into lotions, creams, gels, and tonics. These product forms may be used for a number of applications, including, but not limited to, hand and body lotions, (facial) anti-ageing preparations, make-ups including foundations, sunscreens, sunless tanners and the like. Any additional components required to formulate such products vary with product type and can be routinely chosen by one skilled in the art.

If the cosmetic or dermatological compositions of the present invention are formulated as an aerosol and applied to the skin as a spray-on product, a propellant is added to the composition.

The cosmetic or dermatological compositions according to the present invention can be prepared by conventional methods in the art such as e.g. by admixing the respective neutral core HMO(s) with all the definitions and preferences given herein with the cosmetically acceptable carrier.

The cosmetic or dermatological composition may comprise further ingredients, which may form part of the carrier. Such ingredients are particularly surfactants, emulsifiers, thickeners, and oils. Such suitable surfactants, emulsifiers, thickeners, and oils are well known to a person skilled in the art. The cosmetic or dermatological compositions of the invention (including the carrier) may comprise further conventional (cosmetic) adjuvants and additives, such as preservatives/antioxidants, fatty substances/oils, water, organic solvents, silicones, thickeners, softeners, emulsifiers, antifoaming agents, aesthetic components such as fragrances, surfactants, fillers, anionic, cationic, non-ionic or amphoteric polymers or mixtures thereof, propellants, acidifying or basifying agents, dyes, colourings/colorants, abrasives, absorbents, chelating agents and/ or sequestering agents, essential oils, skin sensates, astringents, pigments or any other ingredients usually formulated into such compositions.

In accordance with the present invention, the compositions according to the invention may also comprise further cosmetically active ingredients conventionally used in cosmetic or dermatological compositions. Exemplary active ingredients encompass skin lightening agents; UV-filters, agents for the treatment of hyperpigmentation; agents for the prevention or reduction of inflammation; firming, moisturizing, soothing, and/ or energizing agents as well as agents to improve elasticity and skin barrier.

If nothing else is stated, the excipients, additives, diluents, etc. mentioned in the following are suitable for the compositions according to the present invention. The necessary amounts of the cosmetic and dermatological adjuvants and additives can, based on the desired product, easily be determined by the skilled person.

The additional ingredients can either be added to the oily phase, the aqueous phase or separately as deemed appropriate. The mode of addition can easily be adapted by a person skilled in the art.

Examples of cosmetic excipients, diluents, adjuvants, additives as well as active ingredients commonly used in the skin care industry which are suitable for use in the cosmetic or dermatological compositions of the present invention are for example described in the International Cosmetic Ingredient Dictionary & Handbook by Personal Care Product Council (http://www.personalcarecouncil.org/), accessible by the online INFO BASE (http://online.personalcarecouncil.org/jsp/Home.jsp), without being limited thereto.

The cosmetically active ingredients useful herein can in some instances provide more than one benefit or operate via more than one mode of action. Of course, one skilled in this art will take care to select the above mentioned optional additional ingredients, adjuvants, diluents and additives and/or their amounts such that the advantageous properties intrinsically associated with the combination in accordance with the invention are not, or not substantially, detrimentally affected by the envisaged addition or additions.

The cosmetic or dermatological compositions according to the present invention are in particular skin care preparations, or functional (anti-ageing) preparations.

Examples of skin care preparations are, in particular, light protective preparations (sun care preparations), anti-ageing preparations, preparations for the treatment of photo-ageing, body oils, body lotions, body gels, treatment creams, skin protection ointments, moisturizing preparations such as moisturizing gels or moisturizing sprays, face and/or body moisturizers, as well as skin lightening preparations.

Examples of functional preparations are cosmetic or dermatological compositions containing active ingredients such as hormone preparations, vitamin preparations, vegetable extract preparations, anti-ageing preparations, and/or antimicrobial (antibacterial or antifungal) preparations without being limited thereto.

The cosmetic or dermatological compositions according to the present invention can be in a wide variety of forms. Non limiting examples include simple solutions (e.g. aqueous, organic solvent, or oil based), emulsion or micro emulsion (in particular of oil-in-water (O/W) or water-in-oil (W/O) type, silicone-in-water (Si/W) or water-in- silicone (W/Si) type, PIT-emulsion, multiple emulsion (e.g. of oil-in-water-in oil (O/W/O) or water-in-oil-in-water (W/O/W) type) or pickering emulsions), as well as solid forms (e.g. hydrogels, alcoholic gels, lipogels, sticks, flowable solids, or amorphous materials).

These product forms may be used for a number of applications, including, but not limited to, gels, creams, ointments, lotions, serums, powder, aerosol sprays or two component dispensing systems.

In a preferred embodiment, the cosmetic or dermatological compositions according to the present invention are emulsions and/or gels. Even more preferably, the cosmetic or dermatological compositions are emulsions which contain an oily phase and an aqueous phase such as in particular O/W, W/O, Si/W, W/Si, O/W/O, W/O/W multiple or a pickering emulsions.

The amount of the oily phase (i.e. the phase containing all oils and fats including the polar oils) present in such emulsions such as in particular O/W, W/O, Si/W, W/Si, O/W/O, W/O/W multiple or a pickering emulsions is preferably at least 10 wt.-%, such as in the range from 10 to 60 wt.-%, preferably in the range from 15 to 50 wt.-%, most preferably in the range from 15 to 40 wt.-%, based on the total weight of the composition.

The oil phase according to the invention preferably comprises oils selected from butylenglykoldicaprylatAdicaprat, propylenglykoldicaprylatZ-dicaprat, dicaprylylether, Ci2-15-Alkylbenzoat, Cis-38-fatty acid triglyceride, dibutyladipate, cyclomethicone, dimethicone, 2-phenylethylbenzoat, isopropyl lauroyl sarkosinate, caprylic/ capric triglyceride as well as mixtures thereof.

The amount of the aqueous phase present in such emulsions is preferably at least 20 wt.-%, such as in the range from 20 to 90 wt.-%, preferably in the range from 30 to 80 wt.-%, most preferably in the range from 30 to 70 wt.-%, based on the total weight of the composition.

Advantageously in all emulsions of the present invention the ratio of oily phase to aqueous phase is selected in the range of 40:60 to 30 to 70.

In one particular advantageous embodiment, the cosmetic or dermatological compositions according to the present invention are in the form of an oil-in-water (O/W) emulsion comprising an oily phase dispersed in an aqueous phase in the presence of an O/W emulsifier. The preparation of such O/W emulsions is well known to a person skilled in the art.

If the cosmetic or dermatological composition according to the invention is an O/W emulsion, then it contains advantageously at least one O/W- or Si/W-emulsifier selected from the list of, glyceryl stearate citrate, glyceryl stearate SE (selfemulsifying), stearic acid, salts of stearic acid, polyglyceryl-3- methylglycosedistearate. Further suitable emulsifiers are phosphate esters and the salts thereof such as cetyl phosphate (e.g. as Amphisol® A from DSM Nutritional Products Ltd.), diethanolamine cetyl phosphate (e.g. as Amphisol® DEA from DSM Nutritional Products Ltd.), potassium cetyl phosphate (e.g. as Amphisol® K from DSM Nutritional Products Ltd.), sodium cetearylsulfate, sodium glyceryl oleate phosphate, hydrogenated vegetable glycerides phosphate and mixtures thereof. Further suitable emulsifiers are sorbitan oleate, sorbitan sesquioleate, sorbitan isostearate, sorbitan trioleate, cetearyl glucoside, lauryl glucoside, decyl glucoside, sodium stearoyl glutamate, sucrose polystearate and hydrated polyisobutene. Furthermore, one or more synthetic polymers may be used as an emulsifier. For example, PVP eicosene copolymer, aery lates/C 10-30 alkyl acrylate crosspolymer, and mixtures thereof.

The at least one O/W, respectively Si/W emulsifier is preferably used in an amount of 0.5 to 10 wt. %, in particular in the range of 0.5 to 6 wt.-%, such as more in particular in the range of 0.5 to 5 wt.-%, such as most in particular in the range of 1 to 4 wt.-%, based on the total weight of the cosmetic or dermatological composition.

Particular suitable O/W emulsifiers to be used in the cosmetic or dermatological compositions according to the invention encompass phosphate ester emulsifiers such as advantageously 8-10 alkyl ethyl phosphate, C9-15 alkyl phosphate, ceteareth-2 phosphate, ceteareth-5 phosphate, ceteth-8 phosphate, ceteth-10 phosphate, cetyl phosphate, C6-10 pareth-4 phosphate, C12-15 pareth-2 phosphate, C12-15 pareth-3 phosphate, DEA-ceteareth-2 phosphate, DEA-cetyl phosphate, DEA-oleth-3 phosphate, potassium cetyl phosphate, deceth-4 phosphate, deceth-6 phosphate and trilaureth-4 phosphate.

A particular suitable O/W emulsifier to be used in the cosmetic or dermatological compositions according to the invention is potassium cetyl phosphate e.g. commercially available as Amphisol® K at DSM Nutritional Products Ltd Kaiseraugst.

Another particular suitable class of O/W emulsifiers are non-ionic self-emulsifying systems derived from olive oil e.g. known as (INCI Name) cetearyl olivate and sorbitan olivate (chemical composition: sorbitan ester and cetearyl ester of olive oil fatty acids) sold under the tradename OLIVEM 1000.

In one particular embodiment, the invention relates to cosmetic or dermatological compositions with all the definitions and preferences given herein in the form of O/W emulsions comprising an oily phase dispersed in an aqueous phase in the presence of an O/W emulsifier wherein the O/W emulsifier is potassium cetyl phosphate. The amount of oily phase in such O/W emulsions is preferably at least 10 wt.-%, more preferably in the range of 10 to 60 wt.-%, most preferably in the range of 15 to 50 wt.- %, such as in the range of 15 to 40 wt.-%, based on the total weight of the composition.

Preferably, the cosmetic or dermatological compositions according to the invention further comprise at least one fatty alcohol (co-emulsifier), such as in particular cetyl alcohol, cetearyl alcohol and/ or behenyl alcohol. The total amount of one or several fatty alcohols on the topical compositions according to the invention is preferably selected in the range of about 0.1 to 10.0 wt.-%, in particular in the range of about 0.5 to 6.0 wt.-% with respect to the total weight of the topical composition.

Preferably, the topical compositions according to the invention comprise a thickener in particular if the topical composition is in the form of an emulsion to assist in making the consistency of a product suitable. Preferred thickeners are aluminiumsilicates, xanthan gum, hydroxypropylmethylcellulose, hydroxyethylcellulose, polyacrylates such as carbopole® (e.g. Carbopole 980, 981 , 1382, 2984, 5984) or mixtures thereof. Further preferred thickeners encompass acrylate/Cw-30 alkyl acrylate copolymers (such as e.g. Pemulen TR 1 , Pemulen TR 2, Carbopol 1328 by. NOVEON) as well as Aristoflex AVC (INCI: Ammonium Acryloyldimethyltaurate/VP Copolymer).

The cosmetic or dermatological compositions according to the present invention advantageously comprise a preservative. When present, the preservative is preferably used in an amount of 0.1 to 2 wt.-%, more preferably in an amount of 0.5 to 1.5 wt.-%, based on the total weight of the composition.

The cosmetic or dermatological compositions according to the invention in general have a pH in the range of 3 to 10, preferably a pH in the range of 4 to 8 and most preferably a pH in the range of 4 to 7.5 such as in the range of 5 to 6.5. The pH can easily be adjusted as desired with suitable acids, such as e.g. citric acid, or bases, such as sodium hydroxide (e.g. as aqueous solution), triethanolamine (TEA Care), Tromethamine (Trizma Base) and Aminomethyl Propanol (AMP-Ultra PC 2000), according to standard methods in the art.

The amount of the cosmetic or dermatological composition to be applied to the skin is not critical and can easily be adjusted by a person skilled in the art. Preferably the amount is selected in the range of 0.1 to 3 mg/cm ² skin, such as preferably in the range of 0.1 to 2 mg/cm ² skin and most preferably in the range of 0.5 to 2 mg/cm ² skin.

Alternatively, the daily dosage of the composition with all the preferences and definitions as given herein to be topically applied is preferably selected in the range of 1 ml, preferably 2 ml, most preferably 5 ml/day.

In all embodiments of the present invention, the cosmetic or dermatological composition according to the present invention is topically administered to the skin of a subject in need thereof for at least 7 days, preferably at least 28 days.

In another preferred embodiment the cosmetic or dermatological composition according to the present invention is applied once or, preferably, twice per day.

The following examples are provided to further illustrate the compositions and effects of the present invention. These examples are illustrative only and are not intended to limit the scope of the invention in any way.

Examples

Short description

The neutral core human milk oligosaccharides LNT and LNnT were evaluated for their effect on collagen type 1. This was done using an in vitro dermal fibroblast collagen assay. It was found that both LNT and LNnT increase collagen expression. For comparative reasons, also another human milk oligosaccharide was tested, i.e. 6’-silalyllactose (6’SL).

Methodology

Normal human dermal fibroblasts were seeded in cell culture medium (Dulbecco's Modified Eagle Medium supplemented with 10% serum and 1% antibiotics) and allowed to adhere in a humidified cell incubator maintaining 37°C and 5% CO2. Afterwards, the cells were starved with culture medium without serum for 16-24 hours to synchronize the cells their cell cycle. The starvation medium was removed and cell culture medium + LNT or LNnT respectively'6’SL at various concentrations was added to the cells. The cells were treated for 72 hours. The cells were then fixated and immunolabeled with antibodies specific for collagen 1. These labeled cells were analyzed using flow cytometry and compared to the untreated ones (medium without LNT respectively LNnT or 6’SL). Table Results: LNT

Table 1. Normal human dermal fibroblasts were treated for 72 hours with several concentrations of LNT, which resulted in the increase of collagen type 1 proteins. Data are normalized to the untreated control (100%) and expressed as the mean ± SD (n = 4).

Table 2: Results LNnT

Table 2. Normal human dermal fibroblasts were treated for 72 hours with several concentrations of LNnT, which resulted in the increase of collagen type 1 proteins. Data are normalized to the untreated control (100%) and expressed as the mean ± SD (n = 4). Table 3: Results 6’SL

Table 3. Normal human dermal fibroblasts were treated for 72 hours with several concentrations of 6’SL, which resulted in a small increase of collagen type 1 proteins at lower concentration but lost significance at concentrations around and above 1 pM. Data are normalized to the untreated control (100%) and expressed as the mean ± SD (n = 4). In comparison to the human milk oligosaccharide according to the present invention, 6’SL has a much smaller effect. At higher concentrations and in contrast to the human milk oligosaccharide according to the present invention this effect also loses its statistical significance (P-value < 0.05) compared to LNT and LNnT.