US20050249691A1

US20050249691A1 - Cosmetic or dermatological preparation comprising a nutrient medium phase

Info

Publication number: US20050249691A1
Application number: US10/967,232
Authority: US
Inventors: Monika Monks; Sybille Ibanez; Carmen Evangelisti; Sven Gohla
Original assignee: Beiersdorf AG
Current assignee: Prairie Group AG
Priority date: 2003-05-24
Filing date: 2004-10-19
Publication date: 2005-11-10
Also published as: EP2340856B2; US8507276B2; EP2340856B1; WO2004103333A1; JP2007500196A; EP1635782A1; US20060182701A1; US8518422B2; EP2340856A1; US20050287182A1; EP1896092A1

Abstract

A cosmetic or dermatological preparation that comprises collagen and/or a derivative thereof, chitosan and/or a derivative thereof, having a degree of acetylation of up to 50%, and glycosaminoglycan and a derivative thereof.

Description

The present application is a continuation of International Application No. PCT/EP2004/005533, filed May 22, 2004, the entire disclosure whereof is expressly incorporated by reference herein, which claims priority under 35 U.S.C. § 119 of German Patent Application Nos. 103 23 510.8, filed May 24, 2003, 103 55 110.7, filed Nov. 24, 2003, and 102004020035.1, filed Apr. 22, 2004.
The invention comprises a cosmetic or dermatological preparation comprising at least one nutrient medium phase for skin cells or corneal cells in combination with an aerogel or hydrogel matrix, containing collagens, chitosans having a degree of acetylation of at least 50% and chondroitin sulfates. The invention further comprises cell culture media as aqueous phase in combination with the gelling matrix described above in synergistic use with polyurethanes which are used for physiological wound healing or scar reduction.
Various circulations exist within the milieu of the human body, such as the blood circulation, the lymphatic system and the intracellular and extracellular tissue fluids. The composition of the solvent “water” with its mineral and bioorganic constituents in these various “transport media” are approximately the same and are based, highly simplified, on salts, amino acids, vitamins, sugars, proteins and proteids, and trace elements. During evolution, our body has learnt to create within these fluids “communication networks” and nutritional strategies, and an equilibrium of catabolic to anabolic processes, which make the complex life of our multicellular body in fact possible. In this environment, our body has learnt to construct from its “single individuals”, the cells, a complicated but efficient network of direct and mediator-related contacts. These “communication pathways” function efficiently and harmlessly only if the natural dynamic equilibrium of our body, the so-called “homeostasis”, exists. If cells are removed from the tissue assemblage or if the homeostasis in the tissue assemblage is impaired, it is no longer possible for individual cells to exist or tissues to function healthily. The medical and biosciences have for decades looked for possibilities of cultivating tissues or individual cells in suitable environmental conditions outside the body. This succeeded only when it was possible to simulate as perfectly as possible the living conditions in the body for the single cells or tissue constituents to be cultivated.
Thus, if cells are removed from intact tissue, they must be cultivated in environments which come as close as possible to the natural living conditions in the body. Requirements for this are supply and transport away of nutrients, and the presence of vital factors.
These environments are well-defined compositions of mineral and biomaterials which are known in science as cell culture media. Cell culture media are obtainable from suitable specialist retailers as powder or liquid media and have slightly different compositions depending on the nature of the cells or tissue constituents to be cultivated. Cell culture media are used in liquid form. With a suitable composition, they make it possible to maintain or even multiply microorganisms or cells in culture, i.e., outside the body.
In the course of the findings in tissue research it was possible to find and investigate the individual needs of cells and cells in intact tissues. In this connection, the ratio of mineral and bioorganic substances of a cell culture medium is slightly variable from cell type to cell type and must be ascertained accurately for optimized survival and growth. The composition of the cell culture medium always depends on the requirements of the cells to be grown. A distinction is made between synthetic media, whose ingredients are accurately known on the basis of pure substances, and complex media, whose exact composition may vary and is in part not accurately known. Cell culture media comprise, besides water, usually a carbon source and a nitrogen source, phosphate compounds and sulfur compounds, and minerals and, optionally, growth promoters or vitamins.
If the compositions of the media are suitable, the cells are able to multiply and produce the factors necessary for survival themselves “in situ”.
In order to generate good growth of the cells, sera are frequently added to the cell culture media. The serum has a complicated composition and provides the cells with, inter alia, hormones, adhesion factors, and amino acids. Culture media which contain sera are, however, costly and do not allow thermal sterilization. It is therefore attempted to make do with media which contain no sera. Serum-free culture media make it possible to cultivate cells under controlled and defined conditions, so that unwanted effects due to variations in the serum composition are eliminated. In addition, contamination of the cell cultures with viruses and bacteria is reduced on use of serum-free media.
It is known that skin cells can be kept alive particularly well preserved and for long periods of time and can even be induced to grow and differentiate in one-, two- and three-dimensional cultures by optimizing the ingredients in the culture medium. It has also been possible to demonstrate that suitable media also make possible the production of growth factors in situ.
When there are extreme changes in the skin resulting from extensive burns or chills, the integrity and the functionality of the cutaneous tissue may be so impaired that it is no longer able to regenerate of its own accord. The body responds to such severe events with hyperthermia, massive release of mediators of inflammation and irritation, and with an enormous loss of fluid, which in the past has actually always inevitably led to the death of people with severe burns. Burns and chills which have led to losses of cutaneous tissue can according to the prior art be compensated by skin transplants and thus the skin can be closed. However, this is successful only if sufficient remaining skin is available for transplantation. In cases of burns of more than 60% of the total cutaneous tissue, transplantation on its own is usually of no assistance. It is necessary to re-produce viable tissue from the remaining skin cells. In this connection, because of the rejection reactions between non-HLA-compatible tissues, it is not possible to take allogeneic skin or allogeneic skin cells. It is therefore necessary to form a new cutaneous tissue in situ from the remaining viable skin cells.
The hornified epidermis forms the protective shield of the skin. For this function to be optimally exercised it is necessary for the skin cells (keratinocytes) to pass through the process of so-called epidermal differentiation. After division of the cells in the basal layer, the keratinocytes migrate to the skin surface and undergo a number of changes during this, until they form the horny layer (stratum corneum) as dead, flat, anuclear corneocytes, and eventually are desquamated. During the epidermal differentiation there is formation of various proteins having specific functions. These include, inter alia, keratins, involucrin, filaggrin and transglutaminase. For optimal formation of the epidermis and the horny layer it is necessary for these proteins to be formed in coordinated fashion and in sufficient quantity.
Many cosmetics, skin care products or wound healing products which help to compensate or at least reduce the disorders of the skin are known in the prior art.
Thus, for example, geroderma is cosmetically treated primarily with vitamin A derivatives or hydroxy acids which lead, via stimulation of the proliferation of the basal cells in the epidermis, to a thickening of the epidermis and thus smoothing of the skin. More recent approaches consist of targeted replacement of the proteins which are absent or present in reduced quantity in dry skin or geroderma, or indirect intervention in the metabolic processes which are disturbed in dry skin or with increasing age, in order to normalize them. An example which may be mentioned here is stimulation of collagen synthesis with the aim of reducing wrinkles. In addition, for example, laminin, substances for prolonging the lifetime of skin cells and certain extracts for stimulating epidermal differentiation are employed. However, some of these are pharmacologically active substances with a high potential for side effects.
None of the preparations known from the prior art on their own enable the skin to regenerate itself without displaying unwanted side effects.
The object of the present invention is to provide a preparation with which it is possible for the skin to be able to regenerate itself without displaying unwanted side effects.
EP 296078 describes the preparation and use of a mixture of collagen, acetylated chitosan with a degree of acetylation of from 10 to a maximum of 40% and glycosylaminoglycan as artificial skin. This mixture of biomaterials is used in orthopedics and in plastic surgery, and as reconstitution matrices for the regeneration of nerve, bone and skin tissues, the latter in particular for very severe burns.
The problem is that it has not heretofore been possible to incorporate the biomaterials described in EP 296078 stably into conventional cosmetic or dermatological matrices. Nor has there been heretofore any provision as cosmetic or as wound care product.
The stated objects are achieved by a cosmetic or dermatological preparation corresponding to the main claim. The dependent claims relate to advantageous embodiments of the preparation of the invention. In addition, the invention comprises the use of such preparations.
It was surprising and could not have been foreseen by one of skill in the art that a cosmetic and/or dermatological preparation which comprises a mixture of collagen, acetylated chitosan with a degree of acetylation not exceeding 50% and glycosylaminoglycan achieves the stated objects. In particular, preparations containing in addition one or more skin cell culture media in which the media is preferably selected from DMEM/HAM's F-12 (1:1) and/or MCDB 153 are to be designated as particularly advantageous. In addition, all culture media which make it possible to culture healthy differentiated skin (3-D models), i.e., in particular, media used to culture primary fibroblasts and/or keratinocytes and moreover make complete reconstitution of the skin possible may be contemplated. Serum substitutes for serum-free cell cultures are also advantageous but not essentially necessary.
It is preferred to add to the preparation of the invention at least one compound, preferably all, selected from the group of L-cystine, L-glutamine, hypoxanthine, L-glutamic acid, thymidine, glycine, lipoic acid, L-histidine HCl, linoleic acid, L-isoleucine, putrescine 2HCl, NaCl, choline chloride, KCl, putrescine, sodium acetate, vitamin B₁₂, Na₂HPO₄, biotin, MgCl₂, calcium pantothenate, CaCl₂, nicotinamide, glucose, pyridoxine HCl, sodium pyruvate, thiamine HCl, NaHCO₃, adenine, phenol red, myo-inositol, HEPES, lipoic acid, thymidine, L-alanine, folic acid, L-arginine HCl, riboflavin, L-asparagine, L-aspartic acid, CuSO₄, L-cysteine HCl, FeSO₄, L-glutamine, MnSO₄, L-glutamic acid, (NH₄)₆Mo₇O₂₄, glycine, NiCl₂, L-histidine HCl, H₂SeO₃, L-isoleucine, Na₂SiO₃, L-leucine, SnCl₂, L-lysine HCl, NH₄VO₃, L-methionine, ZnSO₄, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, insulin human, ZnSO₄, CoCl₂, CuSO₄, Na₂SeO₃, AlCl₃, CrK(SO₄)₂, NiCl₃, MnCl₂, EDTA.Na₂, polysorbate 80, L-leucine, Na₂HPO₄, L-methionine, NaH₂PO₄, L-phenylalanine, MgSO₄, MgCl₂, L-serine, CaCl₂, L-threonine, Fe(NO₃)₃, L-tryptophan, L-tyrosine, α-biotin, Na pyruvate, folic acid, D-Ca pantothenate, L-alanine, L-arginine HCl and/or pyridoxal HCl.
A particularly advantageous combination comprises the linkage of cell-nourishing culture media, preferably media for cultivating skin cultures or corneal cultures of all types with a cellular matrix containing collagens, acetylated chitosans with a degree of acetylation of up to 50%, preferably up to 40%, and chondroitin sulfates. This combination alone, mixed with cosmetic preparations or incorporated into polyurethane matrices proves to be extremely efficient in relation to skin regeneration, skin care and wound healing. The invention makes it possible to regenerate skin or partial skin from individual cells (dermis and epidermis), to transfer this gel matrix precultured in vitro to damaged tissue for complete skin renewal, and the prevention or reduction of scar tissue associated with wound healing. The present invention further provides the ideal environment (matrix) for renewing the skin on topical application.
The preparation of the matrix is described in EP 296078. The disclosure of EP 296078 in its entirety is hereby a constituent of the present invention.
It is novel and was not foreseeable by one of skill in the art, and is thus according to the invention, that it is possible to obtain the matrix described in EP 296078 entirely through marine and or synthetic raw material sources and that the results are the same as in the patent EP 296078.
The preparation of the invention, also described as primary microporous or nanoporous matrix, preferably consists of marine collagens selected from the group of type 3, type 1, type 4 and/or type 5 or blends thereof, chitosans, preferably with a molecular weight of from 80 000 D to 1.5 000 000 D, and with a degree of acetylation of from 5% to 50%, blended with a mixture of chondroitin 4- and 6-sulfates, which are employed at 3 to 15% of the initial amount of the collagens. The preparation or matrix of the invention can be imagined to be in the form of a microtubular or nanotubular sponge.
On lyophilization, the composition of the described molecules generates a nano- or microsponge (matrix).
The matrix consists of an aerogel prepared by lyophilization, which is introduced into the aqueous phase(s), active ingredient phases or media phases of a finished cosmetic or dermatological preparations. In this case, it is converted into a hydrogel, or processed as aerogel into a, or together with a, for example, polyurethane matrix or silicone matrix.
It has been possible to show that the combination according to the invention of marine and/or synthetic collagens, acetylated chitosans and glycosylaminoglycans and, in particular, with the addition of cell culture media makes possible advantages in the morphology and growth rate of primary human keratinocytes and fibroblasts from young and old donors in vitro. In principle, all growth and maintenance media are possible for this, but preferably those adapted to the requirements of skin cells and further enabling the construction of “new skin” from individual dermis or epidermis cells in the described matrix. Application studies show that irritated skin is soothed on treatment with the matrix of the invention. It is particularly advantageous in this connection that the collagen, chitosan and glycosylaminoglycan ingredients of the invention are employed in a balanced ratio to one another, especially as described in EP 296078. This is particularly true because the formation of a micro- or nanotubular aerogel or the retention of this structure in the cosmtic preparation or skin covering takes place only through the specific active ingredients listed according to the invention to one another.
In this case, the stationary biopolymer phase with the disperse phase(s) composed of physiological saline solution, minimal media or complete media is converted into a hydrogel phase. The hydrogel phase with the properties according to the invention results with the matrix components of the invention. The individual or only 2 of the active ingredients alone, or blending of all active ingredients in non-advantageous proportions, do not result in the desired effect, for example, in interplay with the cell culture media as aqueous phase or in interplay with polyurethane matrices.
Preferred is a ratio of collagen to chitosan ingredients of 60 to 90% to 40 to 10%, in particular 75-85% to 25-15%.
Collagens is a designation for a family of long-fiber, linear-colloidal, high molecular weight scleroproteins of the extracellular matrix, which occur in connective tissue (e.g. skin, cartilage, tendons, ligaments, blood vessels), in osseine (the protein-containing base substance of bone) and in dentin together with proteoglycans. They are regarded as the most common animal proteins in terms of quantity, with a proportion of 25-30%. A mutual anchoring of the collagen fibers and of the cells is produced by fibronectin, which is able to bind collagen and other constituents of the extracellular matrix, but also becomes attached to receptors on cell surfaces. The composition of the collagens may vary depending on the origin. Collagens of types 1 to 14 are known, but only types I-III, V and XI have the described fiber structure.
The matrix of the invention may be an ingredient of aqueous gels, emulsions of the O/W, W/O/W or W/O type, microemulsions or cosmetic stick products and can thus be marketed for the first time in conventional cosmetic application forms.
In addition, the invention also relates to the preparation in skin coverings, patches, pads, tissues or bandages. In this regard, particularly in polyurethane-based wound coverings.
One possibility in this connection is a professional wound covering used for the therapy of very severe burn injuries. In-home-use application is likewise possible. In the form of an aerogel, the matrix can be placed on the wound or the part of skin to be treated. The constituents of the matrix of the invention are biological polymers which can, through a specific mixing ratio, be converted into a stable aerogel and even be reconstituted as stable hydrogel. It was possible to show in numerous experiments that this gel matrix produces complete healing skin again from individual skin cells when it is placed appropriately on the wound. Advantages were found and demonstrated experimentally for the formulations of the invention for cell regeneration and proliferation of primary skin cells of the keratinocyte and fibroblast type. It was also possible through the chondroitin sulfate, chitosan and collagen matrix interacting with the skin cells, in particular in the 3-D skin models, to induce the production of elastin, fibrillin and further biomarkers which are responsible for the quality of a healthy skin. It further is possible, through the interplay of matrix molecules, as described above, and the cell culture media, to markedly improve the reticular interlocking of the epidermis in the dermis. It is thus possible, through the matrix model of the invention in interplay with the culture media, to achieve ideal regeneration of complete skin from only a few skin cells, and supply a pre-existing skin with the ideal healthy growth environment and nutrient factors. In interplay with polyurethane components, skin regeneration here can be optimized under semi- or occlusive conditions, which helps to normalize in particular, keloids and other scars.
The process for recovering the constituents of the described matrix consists of adding acetylated chitosan to a collagen/water/optionally cell culture medium solution and subsequently adding glycosylaminoglycan, in particular chondroitin 4- or 6-sulfate.
The proportions according to the invention of the at least 3 active ingredients of the matrix allow sustained or controlled release of active ingredients such as, for example, Q10, retinol, AHA, etc. and, in addition, optionally reduces the side effects of these agents.
One of skill in the art understands by the term skin cell culture medium all liquid, powdered or solid media in or on which individual cells can multiply or be cultivated. One of skill in the art makes a distinction in this connection between purifying media such as, for example, phosphate-buffered saline solution, minimal maintenance media and so-called complete media, in which cells are healthy and metabolically active. Complete media may on the one hand be provided with growth factors from animal sera or so-called synthetic sera substitutes in order to improve the growth of specific cells or make same possible at all. For every cell type, but especially the culture of primary cells, there are media and media blends which support the growth, differentiation or metabolism of the specific cells particularly well. The composition according to the invention of collagens, chitosans and chondroitin sulfates comprises blending with all purifying, minimal or complete media, but especially complete media which have been composed for culturing skin cells and serve in particular as nutrient media for primary human fibroblasts and keratinocytes and which, as nutrient media, make it possible for the dermis to be regenerated from individual fibroblasts, or for the epidermis to be regenerated from individual keratinocytes. The skin cell culture media of the invention are thus particularly suitable for cultivating skin cells. In particular, the skin cell culture media of the invention which are suitable, in the composition of their individual ingredients, for the following purposes are described: culture of fibroblast cells

- culture of keratinocyte cells
- co-cultures of keratinocytes and fibroblasts
- co-cultures of fibroblasts/keratinocytes and further skin-relevant cells such as immune cells, melanocytes etc.
- culture media for generating three-dimensional skin models

The media of the invention act on keratinocyte/fibroblast mixed cultures and 3-D skin models.
It was possible to show, surprisingly, that cosmetic or dermatological preparations containing the mixture according to the invention of biomolecules and skin cell culture media are able in or on the human skin itself to activate or simulate those mechanisms which the skin uses for homeostasis and healthy autopoiesis. It is possible in this connection for mixtures of fibroblast- or keratinocyte-relevant growth media to be employed directly or in suitable vesicle technologies and be used for medical/pharmaceutical purposes and cosmetic purposes.
In particular, so-called serum-free media have proved to be advantageous when the cell fraction of the primary keratinocytes and primary fibroblasts is to be positively influenced in the sense of optimized homeostasis.
The use of the skin-relevant culture media in interplay with the matrix biomolecules brings about autologous, healthy and individual regeneration of deficient skin functions in vitro, ex vivo and in vivo. It is thus possible for regeneration of the skin, skin tautness or else simply only the contribution to skin care to be significantly improved.
In principle, all skin cell culture media are suitable for use in cosmetic preparations. Particularly suitable skin cell culture media are those employed in the literature for cultivating skin cells or skin-relevant cells, for treating skin irritations and burns. In particular, media for cultivating remaining cells after extensive burns show an extremely advantageous effect after application of the topical preparations.
Skin cell culture media advantageous according to the invention are media which permit neogenesis of fibroblasts or keratinocytes alone or in mixed cultures or reduce the formation and passaging of non-benign cells.
The skin cell culture media DMEM/HAM's F-12 (1:1) and MCDB 153 are particularly suitable for the use according to the invention in cosmetic preparations.
According to the reference in the literature from Barnes D. and Sato G.; Anal. Biochem 102, 255 [1980], DMEM/HAM's F-12 (1:1) is a 1:1 mixture where the nutrient content of Ham's F-12 medium is increased through addition of Dulbecco's MEM (DMEM=Dulbesccos Modified Eagles medium). This medium is the basis for cultivating cell lines for human proteins such as, for example, erythropoetin.

DMEM/HAM's F-12 (1:1) has the following composition (data in mg/L):



NaCl	6999.5	L-Leucine	59
KCl	311.8	L-Lysine HCl	91.25
Na₂HPO₄	71	L-Methionine	17.24
NaH₂PO₄—H₂O	62.5	L-Phenylalanine	35.5
MgSO₄-7H₂O	100	L-Proline	17.25
MgCl₂-6H₂O	61	L-Serine	26.25
CaCl₂	116.61	L-Threonine	53.5
Fe(NO₃)₃-9H₂O	0.05	L-Tryptophan	9
FeSO₄-7H₂O	0.417	L-Tyrosine	38.7
CuSO₄-5H₂O	0.00125	L-Valine	52.85
ZnSO₄-7H₂O	0.432	Choline chloride	9
D-Glucose	3151	α-Biotin	0.00365
NaHCO₃	2438	Folic acid	2.65
Na Pyruvate	55	D-Ca pantothenate	2.24
Phenol red	12.5	Nicotinamide	2.02
myo-Inositol	12.6	Pyridoxcal HCl	2
L-Alanine	4.5	Pyridoxine HCl	0.031
L-Arginine HCl	147.5	Riboflavin	0.22
L-Asparagine-H₂O	7.5	Thiamine HCl	2.17
L-Aspartic acid	6.65	Vitamin B₁₂	0.68
L-Cysteine HCl	15.75	Hypoxanthin	2.05
L-Cystine	24	Thymidine	0.37
L-Glutamine	365.3	Lipoic acid	0.11
L-Glutamic acid	7.35	Linoleic acid	0.042
Glycine	18.75	Putrescine 2HCl	0.081
L-Histidine HCl—H₂O	31.5
L-Isoleucine	54.5

MCDB 153 is employed, according to the reference in the literature Barnes D. and Sato G.; Anal. Biochem 102, 255 [1980], for cultivating human keratinocytes. Further, as minimal medium PBS, phosphate-buffered saline, with pH values of from 3.5 to 8.

MCDB 153 has the following composition (mg/L):



NaCl	7599	Choline chloride	13.96
KCl	111.83	Putrescine	0.1611
Sodium acetate-3H₂O	500	Vitamin B₁₂	4.07
Na₂HPO₄-7H₂O	536.2	Biotin	0.0146
MgCl₂-6H₂O	122	Calcium pantothenate	0.258
CaCl₂-2H₂O	4.411	Nicotinamide	0.03663
Glucose	1081	Pyridoxine HCl	0.06171
Sodium pyruvate	55	Thiamine HCl	0.3373
NaHCO₃	1176	Adenine	24.32
Phenol red	1.317	myo-Inositol	18.02
HEPES	6600	Lipoic acid	0.2063
Thymidine	0.7266	Folic acid	0.79
L-Alanine	8.91	Riboflavin	0.03764
L-Arginine-HCl	210.7	CuSO₄-5H₂O	0.0002496
L-Asparagine	15.01	FeSO₄-7H₂O	1.39
L-Aspartic acid	3.99	MnSO₄-5H₂O	0.000241
L-Cysteine HCl—H₂O	42.04	(NH₄)₆Mo₇O₂₄-4H₂O	0.001236
L-Glutamine	877.2	NiCl₂-6H₂O	0.0001188
L-Glutamic acid	14.71	H₂SeO₃	0.003869
Glycine	7.51	Na₂SiO₃-9H₂O	0.1421
L-Histidine HCl—H₂O	16.77	SnCl₂-2H₂O	0.0001128
L-Isoleucine	1.968	NH₄VO₃	0.000585
L-Leucine	65.6	ZnSO₄-7H₂O	0.144
L-Lysine-HCl	18.27
L-Methionine	4.476
L-Phenylalanine	4.956
L-Proline	34.53
L-Serine	63.06
L-Threonine	11.91
L-Tryptophan	3.06
L-Tyrosine	2.718
L-Valine	35.13

The advantage of the DMEM/HAM's F-12 (1:1) and MCDB 153 media is that they are particularly selected and suitable in cosmetic or dermatological preparations for the cultivation of monolayer, two-dimensional and organotypical skin models, and permit the in vitro and ex vivo stimulation or retention of skin-specific biofunctions.

It is additionally advantageously possible for the media to have solutions of the following composition A or B added thereto as serum substitutes.



	Solution A	Solution B
	Components (1000x)	μM

	FeSO₄-7H₂O	3000
	ZnSO₄-7H₂O	3000
	CoCl₂-6H₂O	1000
	CuSO₄-5H₂O	10
	Na₂SeO₃	10
	AlCl₃-6H₂O	5
	CrK(SO₄)₂-12H₂O	1.4
	NiCl₃-6H₂O	1
	MnCl₂-4H₂O	1
	EDTA.Na₂-2H₂O	30000
	Polysorbate 80 VG	3820
	Insulin human in 0.01 M HCl	86

The liquid media are usually, according to statements in the literature, prepared by using high-purity, pyrogen-free water, this complies with the WFI quality (water for injection) of Pharmacopeia Europa. The liquid media are sterilized by filtration and bottled, the systems and methods of manufacture being such that entry of endotoxins and microbes is substantially precluded.
The media of the invention show advantageous properties in relation to skin regeneration even if the media compositions are altered, such as, for example, with or without choline chloride, with or without H₂SeO₃.
The skin cell culture media and the mixture of biomolecules (collagen/chitosan/-glycosylaminoglycan) and, optionally, additives are mixed into cosmetic or dermatological preparations to give a proportion of up to 99.9% by weight based on the total mass of the preparation.
By cosmetic or dermatological preparations or matrices are meant topical preparations which are suitable for applying said media to the skin in fine distribution and preferably in a form which can be absorbed through the skin. Examples suitable for this purpose are aqueous and hydroalcoholic solutions, sprays, foams, foam aerosols, ointments, aqueous gels, emulsions of the O/W or W/O type, microemulsions, hydrophilic or lipophilic patches or cosmetic stick products. Particularly preferably suitable as carrier is an aqueous gel, an O/W emulsion, a W/O/W emulsion or a microemulsion. The preparation can also be used for the purposes of the invention in body-cleansing compositions such as, for example, soaps, shower baths, shampoos and the like.
The cosmetic formulations are in particular hydrogels or emulsions of any type, preferably O/W emulsions.
All lipids known in cosmetics can be employed as oily or lipid phase.
Preparations of the invention in the form of an emulsion contain one or more emulsifiers. These emulsifiers may advantageously be chosen from the group of nonionic, anionic, cationic or amphoteric emulsifiers.
Besides water and physiologically suitable solvents, it is possible to use, inter alia, care constituents, oils, waxes, fats, refatting substances, thickeners, antioxidants, emulsifiers, substances suitable as sunscreen filters, enzymes, amino acids, proteins, polysaccharides and/or fragrances. According to the invention, apart from the aforementioned substances the preparations contain optionally the additives customary in cosmetics, for example perfume, dyes, antimicrobial substances, refatting agents, complexing and sequestering agents, pearlescent agents, plant extracts, vitamins, active ingredients, preservatives, bactericides, pigments which have a coloring effect, thickeners, emollient, moisturizing and/or humectant substances, or other usual ingredients of a cosmetic or dermatological formulation such as alcohols, polyols, polymers, foam stabilizers, electrolytes, organic solvents or silicone derivatives.
Suitable preparations are also those which can be employed for professional wound management and wound healing or reduction of surgical scars or the like, such as, for example, polyurethane preparations in combination with chitosan/collagen/chondroitin 6-sulfate sponges or solutions.
Additions which prove to be advantageous are specific active ingredients such as, for example, of antioxidants. The added antioxidants are advantageously selected from the group of amino acids (e.g. glycine, lysine, arginine, cysteine, histidine, tyrosine, tryptophan) and derivatives thereof (as salt, ester, ether, sugar, nucleotide, nucleoside, peptide and lipid compound), imidazoles (e.g. urocanic acid) and derivatives thereof (as salt, ester, ether, sugar, nucleotide, nucleoside, peptide and/or lipid compound), peptides such as D,L-carnosine, D-carnosine, L-carnosine, anserine and derivatives thereof (e.g. as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and lipid compound), carotenoids, carotenes (e.g. α-carotene, β-carotene, ψ-lycopene, phytoene,) and derivatives thereof (e.g. as salt, ester, ether, sugar, nucleotide, nucleoside, peptide and/or lipid compound), chlorogenic acid and derivatives thereof (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and/or lipid compound), aurothioglucose, propylthiouracil and other thiols (e.g. thioredoxin, lipoic acid, glutathione, cysteine, cystine, cystamine and their glycosyl, N-acetyl, methyl, ethyl, propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters) and the salts thereof, dilauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and derivatives thereof (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and/or lipid compound) and sulfoximine compounds (e.g. homocysteine sulfoximine, buthionine sulfones, penta-, hexa-, heptathionine sulfoximine) in very low tolerated dosages (e.g. pmol to μmol/kg). Also (metal) chelators (e.g. apoferritin, desferral, lactoferrin, α-hydroxy fatty acids, palmitic acid, phytic acid) and derivatives thereof (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and/or lipid compound), α-hydroxy acids (e.g. citric acid, lactic acid, malic acid), humic acid, bile acid, bile extracts, bilirubin, biliverdin, EDTA, EGTA and derivatives thereof, unsaturated fatty acids and derivatives thereof (e.g. γ-linolenic acid, linoleic acid, oleic acid), folic acid and derivatives thereof, furfurylidenesorbitol and derivatives thereof, ubiquinone, ubiquinol, plastoquinone and derivatives thereof (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and lipid compound), vitamin C and derivatives (e.g. ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate), tocopherols and derivatives (e.g. vitamin E acetate), and phenolic compounds and plant extracts containing same such as, for example, flavonoids (e.g. glycosylrutin, ferulic acid, caffeic acid), furfurylideneglucitol, butylated hydroxytoluene, butylated hydroxyanisole, nordihydroguaiaretic resin acid, nordihy-droguaiaretic acid, trihydroxybutyrophenone and derivatives thereof (as salt, ester, ether, sugar, nucleotide, nucleoside, peptide and lipid compound), uric acid and derivatives thereof, mannose and derivatives thereof (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and lipid compound), zinc and its derivatives (e.g. ZnO, ZnSO₄), selenium and its derivatives (e.g. selenomethionine, ebselen), stilbenes and derivatives thereof (e.g. stilbene oxide, trans-stilbene oxide) and the derivatives suitable according to the invention (as salt, ester, ether, sugar, thiol, nucleotide, nucleoside, peptide and/or lipid compound) of these active ingredients mentioned.
An additional use of a buffer may become necessary in order to stabilize the ingredients of the disperse phase. Phosphate-buffered saline solutions or citrate buffers are preferably employed in this case.
Besides the antioxidants or from the group thereof, combinations of the preparations of the invention with specific ingredients which are preferably chosen from the group of Q10, AGR, Zn orotrate, carnitine, creatine and/or taurine are favored.
Areas of application of the preparation of the invention which have proved to be particularly advantageous are the care of all skin types with the exception of all septic inflammations, also special applications such as microdermal abrasion, acid peeling and retinol treatments. The skin regeneration and soothing of the skin by the preparations of the invention is detectable in these cases.
Additionally preferred is cosmetic care of the skin, in particular for beautification.
The packaging can include all cosmetically customary dosage systems such as, for example, jars, pump bottles, pipette bottles, cartridges or capsules.
For problematic formulations into which the cell medium cannot be incorporated, it would be possible to mix the cell culture media and cosmetic product only before use, through special packaging elements such as, for example, double cartridges with mixing head as known, for example, from 2-component adhesives. The packaging of the cell culture medium might also be designed for refilling, so that only fresh product is used.
It is advantageous to incorporate the matrix of the invention in polyurethane matrices and configure them as cosmetic skin covering, wound covering in plasters or bandages or as pad. Possible polyurethane matrices which should be mentioned in particular are those from the publications DE 42 33 289, DE 43 08 347, DE 43 08 445, DE 43 28 190 and DE 101 28 685, which are hereby included in the disclosure of the present invention.
Advantageous exemplary embodiments of the present invention follow. The quantitative data are always based on weight % as long as nothing contrary is indicated. It is possible in all the preparations for the ratio described above of the matrix molecules collagen, chitosan and glycosylaminoglycan to be from 0.00001% by weight to 99% by weight of the final formulation, preferably 0.0005% by weight to 50% by weight and ideally 0.0015 to 30% by weight, based on the total mass of the preparation. The dispersant “culture medium” should correspond to an osmotic pressure of a 0.5 to 2% sodium chloride solution, but ideally correspond to the physiological osmotic pressure of human tissue, especially of the skin.

EXAMPLES

Example 1



	Formula (INCI/CTFA adopted names)	% w/w

	WATER (AQUA)	66.9268
	TRIISOSTEARIN	4.0000
	BUTYLENE GLYCOL	3.0209
	PETROLATUM	2.7000
	GLYCERIN	2.5800
	CETEARYL ETHYLHEXANOATE	2.2500
	HYDROGENATED COCO-GLYCERIDES	2.0000
	CAPRYLIC/CAPRIC TRIGLYCERIDE	2.0000
	CETEARYL ALCOHOL	1.6000
	OLEA EUROPAEA (OLIVE) OIL	1.5000
	UNSAPONIFIABLES
	PEG-30 STEARATE	1.1000
	TOCOPHERYL ACETATE	1.0000
	STEARIC ACID	1.0000
	SORBETH-30	1.0000
	PALMITIC ACID	1.0000
	CYCLOMETHICONE	1.0000
	PHENOXYETHANOL	0.7556
	STEARYL ALCOHOL	0.5880
	POLOXAMER 188	0.5000
	BEESWAX (CERA ALBA)	0.5000
	CETEARETH-20	0.4000
	SODIUM CARBOMER	0.3300
	LANOLIN ALCOHOL	0.3000
	CETYL ALCOHOL	0.2640
	ISOPROPYL MYRISTATE	0.2500
	GLYCERYL POLYMETHACRYLATE	0.2150
	CETEARETH-25	0.2040
	METHYLPARABEN	0.1526
	LECITHIN	0.1490
	HISTIDINE	0.1002
	SODIUM CITRATE	0.1000
	DISODIUM EDTA	0.1000
	CETEARETH-15	0.0900
	GLUCOSYLRUTIN	0.0850
	BUTYLPARABEN	0.0417
	ETHYLPARABEN	0.0402
	MYRISTYL ALCOHOL	0.0360
	ISOBUTYLPARABEN	0.0204
	MYRETH-4	0.0180
	ASCORBYL PALMITATE	0.0175
	SOLUBLE COLLAGEN	0.0171
	ISOQUERCITRIN	0.0150
	PROPYLPARABEN	0.0110
	PROPYLENE GLYCOL	0.0050
	CHITOSAN	0.0046
	TOCOPHEROL	0.0035
	SODIUM CHLORIDE	0.0027
	SODIUM CHONDROITIN SULFATE	0.0019
	GLUCOSE	0.0019
	LYSINE HYDROCHLORIDE	0.0006
	THREONINE	0.0004
	ARGININE	0.0004
	SERINE	0.0002
	POTASSIUM CHLORIDE	0.0002
	TRYPTOPHAN	0.0001
	MAGNESIUM SULFATE	0.0001
	GLYCINE	0.0001
	CALCIUM CHLORIDE	0.0001
	FOLIC ACID	0.0001
	CALCIUM PANTOTHENATE	0.0001

The components HISTIDINE, GLUCOSE and CALCIUM CHLORIDE therein are the components of the aqueous phase which serves as disperse phase.

Example 2



	Formula (INCI/CTFA adopted names)	% w/w

	WATER (AQUA)	71.1509
	GLYCERIN	4.3000
	METHYLGLUCOSE SESQUISTEARATE	3.2200
	TOCOPHERYL ACETATE	2.0050
	PPG-15 STEARYL ETHER	2.0000
	ETHYLHEXYL METHOXYCINNAMATE	2.0000
	PANTHENOL	1.8750
	TRIISOSTEARIN	1.7000
	SORBITAN STEARATE	1.3800
	ETHYLHEXYL PALMITATE	1.3000
	ISOPROPYL STEARATE	1.2000
	CAPRYLIC/CAPRIC TRIGLYCERIDE	1.2000
	MACADAMIA TERNIFOLIA SEED OIL	1.0000
	DIMETHICONE	0.8000
	PHENOXYETHANOL	0.7556
	MYRISTYL MYRISTATE	0.6000
	OCTYLDODECANOL	0.5470
	CYCLOMETHICONE	0.5000
	GLYCERYL POLYMETHACRYLATE	0.4300
	IRVINGIA GABONENSIS KERNEL BUTTER	0.3100
	IMIDAZOLIDINYL UREA	0.3000
	XANTHAN GUM	0.1700
	TITANIUM DIOXIDE	0.1550
	METHYLPARABEN	0.1571
	HYDROGENATED COCO-GLYCERIDES	0.1350
	DISODIUM EDTA	0.1000
	BHT	0.1000
	ACRYLATES/C10-30 ALKYL ACRYLATE	0.1000
	CROSSPOLYMER RETINYL PALMITATE	0.0990
	TROMETHAMINE	0.0700
	CETEARYL ALCOHOL	0.0500
	BUTYLPARABEN	0.0447
	ETHYLPARABEN	0.0402
	LECITHIN	0.0350
	SODIUM PCA	0.0250
	BUTYLENE GLYCOL	0.0209
	ISOBUTYLPARABEN	0.0204
	ALUMINA	0.0200
	SOLUBLE COLLAGEN	0.0171
	ASCORBYL PALMITATE	0.0125
	PROPYLPARABEN	0.0111
	PROPYLENE GLYCOL	0.0100
	SILICA	0.0088
	SODIUM POLYACRYLATE	0.0075
	CHITOSAN	0.0046
	TOCOPHEROL	0.0035
	SODIUM CHLORIDE	0.0027
	SODIUM CHONDROITIN SULFATE	0.0019
	GLUCOSE	0.0019
	LYSINE HYDROCHLORIDE	0.0006
	THREONINE	0.0004
	ARGININE	0.0004
	SERINE	0.0002
	POTASSIUM CHLORIDE	0.0002
	HISTIDINE	0.0002
	TRYPTOPHAN	0.0001
	MAGNESIUM SULFATE	0.0001
	GLYCINE	0.0001
	CALCIUM CHLORIDE	0.0001
	FOLIC ACID	0.0001
	CALCIUM PANTOTHENATE	0.0001

Example 3



	Formula (INCI/CTFA adopted names)	% w/w

	WATER (AQUA)	50.5510
	OCTOCRYLENE	10.0000
	GLYCERIN	7.5000
	CETEARYL ALCOHOL	3.1000
	C12-15 ALKYL BENZOATE	3.0000
	TITANIUM DIOXIDE	2.4000
	TOCOPHERYL ACETATE	2.0000
	DIETHYLHEXYL BUTAMIDO TRIAZONE	2.0000
	CETEARETH-20	2.0000
	BIS-ETHYLHEXYLOXYPHENOL	2.0000
	METHOXYPHENOL TRIAZINE
	HYDROGENATED COCO-GLYCERIDES	1.0000
	GLYCERYL STEARATE	0.7000
	PROPYLENE GLYCOL	0.6250
	PEG-40 CASTOR OIL	0.6000
	PHENOXYETHANOL	0.5180
	FUMARIA OFFICINALIS (FUMITORY) EXTRACT	0.5000
	CITRUS MEDICA LIMONUM (LEMON) EXTRACT	0.5000
	MICA	0.3240
	SODIUM CETEARYL SULFATE	0.3000
	FRAGRANCE (PERFUME)	0.3000
	DMDM HYDANTOIN	0.2850
	XANTHAN GUM	0.2000
	TRISODIUM EDTA	0.1990
	FUMARIC ACID	0.1250
	METHYLPARABEN	0.1050
	TRIMETHOXYCAPRYLYLSILANE	0.1000
	ETHYLPARABEN	0.0280
	BUTYLPARABEN	0.0280
	IODOPROPYNYL BUTYLCARBAMATE	0.0150
	ISOBUTYLPARABEN	0.0140
	PROPYLPARABEN	0.0070
	IRON OXIDES	0.4200
	TITANIUM DIOXIDE	0.2400
	IRON OXIDES	0.1960
	ULTRAMARINES	0.0800
	IRON OXIDES	0.0400
	Collagen	0.03
	Chitosan	0.01
	Chondroitin sulfate	0.005
	ARGININE	0.0004
	SERINE	0.0002
	POTASSIUM CHLORIDE	0.0002
	HISTIDINE	0.0002
	TRYPTOPHAN	0.0001
	MAGNESIUM SULFATE	0.0001
	GLYCINE	0.0001
	CALCIUM CHLORIDE	0.0001
	FOLIC ACID	0.0001

Example 4

O/W Emulsion



KERATINOCYTE MEDIUM MCDB153	40% by weight
COLLAGEN/CHITOSAN/CHONDROITIN SULFATE	6% by weight
matrix and blended in any proportion
WATER (AQUA)
GLYCERIN
HYDROGENATED COCO-GLYCERIDES
SQUALANE
GLYCERYL STEARATE CITRATE
CAPRYLIC/CAPRIC TRIGLYCERIDE
ETHYLHEXYL COCOATE
MYRISTYL ALCOHOL
BUTYROSPERMUM PARKII (SHEA BUTTER)
BUTYLENE GLYCOL
CETYL ALCOHOL
TOCOPHERYL ACETATE
PHENOXYETHANOL
SODIUM CHLORIDE
IMIDAZOLIDINYL UREA
CARBOMER
XANTHAN GUM
METHYLPARABEN
EDTA
SODIUM HYDROXIDE
BHT
ETHYLPARABEN
BUTYLPARABEN
ISOBUTYLPARABEN
PROPYLPARABEN

Example 5

W/O/W Emulsion



	% by weight

PEG-100 stearate	2.00%
Glyceryl stearate	4.00%
Squalane	1.50%
Squalene	1.50%
Isopropyl palmitate	5.40%
MCDB 153/DME 1:1	0.360%
Magnesium sulfate	0.240%
Preservative	0.50%
COLLAGEN/CHONDROITIN SULFATE/CHITOSAN	5%
Water VES	ad 100.00

The fatty phase containing the emulsifier is heated to 80° C., likewise the aqueous phase without that proportion that contains the medium. The two phases are combined at 80° C., homogenized for about 3-10 minutes and then cooled to 48° C. or room temperature. Then, keeping the temperature constant to ±1° C., the proportion of water with medium is added and mixed.

Example 6



	PEG-40 stearate	1.00%
	Glyceryl stearate	2.00%
	Cetyl alcohol	3.00%
	Mineral oil DAB 9	2.00%
	Safflower oil	2.00%
	Isopropyl palmitate	4.50%
	Glycerin	3.00%
	Magnesium sulfate	1.20%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	2.00%
	Deionized water	ad 100.00%
	of which DMEM/HAM's F-12 (1:1)	2.5%

Example 7



	PEG-80 stearate	2.00%
	Cetyl alcohol	3.00%
	Mineral oil DAB 9	1.50%
	Evening primrose oil	2.50%
	Isopropyl palmitate	5.40%
	Propylene glycol	3.00%
	Potassium chloride	0.60%
	Preservative	0.50%
	Collagen/chitosan/gycosylaminoglycan matrix	1.50%
	Water VES	ad 100.00%
	of which DMEM/HAM's F-12 (1:1)	5%

Example 8



	Steareth-100	2.00%
	Myristyl alcohol	1.00%
	Mineral oil DAB 9	3.00%
	Castor oil	3.00%
	Cyclomethicone	2.00%
	Propylene glycol	3.00%
	Glycerin	5.00%
	Potassium chloride	3.00%
	Collagen/chitosan/glycosylaminoglycan matrix	4.50%
	Preservative	0.50%
	Water VES	ad 100.00%
	of which MCDB 153	0.5%

Example 9



	Steareth-20	2.00%
	Cetearyl	3.00%
	Petrolatum	0.50%
	Wheat germ oil	1.50%
	Dimethicone	5.00%
	Glycerin	5.00%
	Sodium chloride	3.00%
	Preservative	0.50%
	PUR	1.50%
	Collagen/chitosan/glycosylaminoglycan matrix	5.65%
	Water VES	ad 100.00%
	of which DMEM/HAM's F-12 (1:1)	15%

Example 10



	Dimethicone copolyol	2.00%
	Cetearyl alcohol	3.00%
	Petrolatum	0.50%
	Wheat germ oil	1.50%
	Dimethicone	5.00%
	Glycerin	5.00%
	Sodium chloride	3.00%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	6.05%
	Water VES	ad 100.00%
	of which MCDB 153	1.5%

Example 11



	PEG-20 behenate	2.00
	Stearyl alcohol	3.00%
	Petrolatum	1.00%
	Grape seed oil	3.00%
	Dimethicone	3.00%
	Sorbitol	5.00%
	Zinc sulfate	3.00%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	10.5%
	Water VES	ad 100.00%
	of which MCDB 153	5%

Example 12



	Decaglyn 1-IS	2.00%
	Stearyl alcohol	3.00%
	Petrolatum	1.00%
	Grape seed oil	3.00%
	Dimethicone	3.00%
	Sorbitol	5.00%
	Zinc sulfate	3.00%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	25.00%
	Water VES	ad 100.00%
	of which MCDB 153	40%

Example 13



	PEG-20 myristate	2.00%
	Stearyl alcohol	3.00%
	Petrolatum	2.00%
	Castor oil	5.00%
	Dimethicone	5.00%
	Sorbitol	5.00%
	Zinc sulfate	3.00%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	3.5%
	Water VES	ad 100.00%
	of which MCDB 153/RPMI 1640/serum substitutes	0.1%

Example 14



	Sucrose laurate	2.00%
	Stearyl alcohol	3.00%
	Petrolatum	2.00%
	Castor oil	5.00%
	Dimethicone	5.00%
	Sorbitol	5.00%
	Zinc sulfate	3.00%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	8.50%
	Water VES	ad 100.00%
	of which MCDB 153	12%

Example 15



	PEG-80 behenate	2.00%
	Glyceryl behenate	4.00%
	Squalane	3.00%
	Castor oil	5.40%
	Glycerin	6.00%
	Magnesium sulfate	2.60%
	Preservative	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	0.0655
	Water VES	ad 100.00%
	of which MCDB 153/DME	8.5%

Example 16

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Octyldodecanol	1.00
	Caprylic/capric triglyceride	1.00
	Dicaprylyl ether	1.00
	Carbomer	0.15
	Glycerin	3.00
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants, etc.
	Collagen/chitosan/glycosylaminoglycan matrix	4.0%
	Water	ad 100.00
	of which DMEM/HAM's F-12 (1:1)	2.5%
	pH adjusted to	5.5

Example 17

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Octyldodecanol	0.25
	Caprylic/capric triglyceride	0.25
	Dicaprylyl ether	0.25
	Carbomer	0.15
	Glycerin	3.00
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants, etc.
	Collagen/chitosan/glycosylaminoglycan matrix	5.0%
	Water	ad 100.00
	of which MCDB 153	0.5%
	pH adjusted to	5.5

Example 18

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Behenyl alcohol	1.00
	Dimethicone	1.50
	Cycolmethicone	1.50
	Carbomer	0.15
	Glycerin	6.00
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants, etc.
	Collagen/chitosan/glycosylaminoglycan matrix	7.5%
	Water	ad 100.00
	of which DMEM/HAM's F-12 (1:1)	5%
	pH adjusted to	5.5

Example 19

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Octyldodecanol	0.25
	Caprylic/capric triglyceride	0.25
	Dicaprylyl ether	0.25
	Dimethicone	0.50
	Carbomer	0.15
	Glycerin	3.00
	Aluminum starch octenyl succinate	0.50
	Talc	0.50
	Bentonite	0.10
	Collagen/chitosan/glycosylaminoglycan matrix	0.40
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153	80%
	pH adjusted to	5.5

Example 20

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Cetyl alcohol	1.00
	Squalane	1.00
	Jojoba oil	1.00
	Liquid paraffin	1.00
	Carbomer	0.10
	Glycerin	3.00
	Serine	0.50
	Tocopherol acetate	1.00
	Carbomer	0.10
	Xanthan gum	0.10
	Collagen/chitosan/glycosylaminoglycan matrix	1.0%
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153	75.5%
	pH adjusted to	6.0

Example 21

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Cetyl alcohol	0.50
	Octyldodecanol	0.40
	Caprylic/capric triglyceride	0.40
	Dicaprylyl ether	0.40
	Carbomer	0.10
	Glycerin	3.00
	Serine	0.50%
	Collagen/chitosan/glycosylaminoglycan matrix	1.2%
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153	45.5%
	pH adjusted to	5.5

Example 22

Emulsion Make Up



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Dimethicone	0.50
	Glycerin	1.50
	1,3 Butylene glycol	1.50
	Magnesium silicate	1.00
	Mica	1.00
	Iron oxides	1.00
	Titanium dioxide	2.50
	Talc	5.00
	Carbomer	0.15
	Collagen/chitosan/glycosylaminoglycan matrix	0.50
	Perfume, preservative, NaOH,	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153/Ham's F12/RPMI 1640	0.5%
	pH adjusted to	5.5

Example 23

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Octyldodecanol	0.25
	Caprylic/capric triglyceride	0.25
	Dicaprylyl ether	0.25
	Octyl methoxycinnamate	4.00
	Benzophenone-3	3.00
	Octyl salicylate	3.00
	Carbomer	0.15
	Glycerin	3.00
	Collagen/chitosan/glycosylaminoglycan matrix	1.75
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153	40%
	pH adjusted to	5.5

Example 24

O/W Emulsion



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Octyldodecanol	0.50
	Caprylic/capric triglyceride	0.50
	Dicaprylyl ether	0.50
	Distarch phosphate	1.00
	Ethanol	10.00
	Carbomer	0.15
	Glycerin	3.00
	Collagen/chitosan/glycosylaminoglycan matrix	0.0625%
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which MCDB 153	35%
	pH adjusted to	5.5

Example 25

Emulsifier Gel



	% by weight

	Glyceryl stearate citrate	3.00
	Stearyl alcohol	1.00
	Ethanol	2.00
	Aluminum starch octenyl succinate	0.25
	Talc	0.25
	Tapioca starch	0.25
	Carbomer	0.15
	Glycerin	3.00
	Collagen/chitosan/glycosylaminoglycan matrix	0.10
	Perfume, preservative, NaOH	q.s.
	dyes, antioxidants etc.
	Water	ad 100.00
	of which DMEM/HAM's F-12 (1:1)	3.5%
	pH adjusted to	5.5

Claims

1-26. (canceled)

27. A cosmetic or dermatological preparation comprising

(a) at least one of collagen and a derivative thereof;

(b) at least one of chitosan and a derivative thereof, having a degree of acetylation of up to 50%; and

(c) at least one of a glycosaminoglycan and a derivative thereof.

28. The preparation of claim 27, wherein at least one of components (a) and (b) is of marine origin.

29. The preparation of claim 27, wherein at least one of components (a) and (b) is of synthetic origin.

30. The preparation of claim 27, wherein the collagen comprises one or more collagens of type 1, type 3, type 4 and type 5.

31. The preparation of claim 27, wherein the chitosan comprises chitosan obtained from at least one of crustaceans and insects.

32. The preparation of claim 27, wherein the chitosan comprises chitosan having a molecular weight of from 80,000 g/mol to 15,000,000 g/mol.

33. The preparation of claim 27, wherein the glycosaminoglycan comprises at least one of chondroitin 4-sulfate and chondroitin 6-sulfate.

34. The preparation of claim 27, wherein the preparation comprises components (a) to (c) in a concentration of from 0.00001% to 99% by weight, based on a total weight of the preparation.

35. The preparation of claim 34, wherein the preparation comprises from 0.0005% to 50% by weight of components (a) to (c).

36. The preparation of claim 35, wherein the preparation comprises from 0.0015% to 30% by weight of components (a) to (c).

37. The preparation of claim 27, wherein the preparation further comprises at least one substance selected from L-cystine, L-glutamine, hypoxanthine, L-glutamic acid, thymidine, lipoic acid, linoleic acid, L-isoleucine, putrescine 2HCl, NaCl, choline chloride, KCl, putrescine, sodium acetate, vitamin B₁₂, Na₂HPO₄, biotin, calcium pantothenate, CaCl₂, nicotinamide, glucose, pyridoxine HCl, sodium pyruvate, thiamine HCl, NaHCO₃, adenine, phenol red, myo-inositol, HEPES, L-alanine, folic acid, L-arginine HCl, riboflavin, L-asparagine, L-aspartic acid, L-cysteine HCl, FeSO₄, MnSO₄, (NH₄)₆Mo₇O₂₄, glycine, NiCl₂, L-histidine HCl, H₂SeO₃, Na₂SiO₃, SnCl₂, L-lysine HCl, NH₄VO₃, L-proline, L-threonine, L-tryptophan, L-tyrosine, L-valine, insulin human, ZnSO₄, CoCl₂, CuSO₄, Na₂SeO₃, AlCl₃, CrK(SO₄)₂, NiCl₃, MnCl₂, EDTA.Na₂, polysorbate 80, L-leucine, L-methionine, NaH₂PO₄, L-phenylalanine, MgCl₂, L-serine, Fe(NO₃)₃, L-tyrosine, α-biotin, D-Ca pantothenate, and pyridoxal HCl.

38. The preparation of claim 27, wherein the preparation comprises components (a) to (c) as at least one of a nanosponge matrix and a microsponge matrix which is reconstituted in cell culture media.

39. The preparation of claim 27, wherein a ratio of collagen and chitosan is 60-90% to 40-10%.

40. The preparation of claim 39, wherein the ratio is 75-85% to 25-15%.

41. The preparation of claim 38, wherein the cell culture media is selected from physiological saline solution, nutrient medium and complete medium for culturing healthy, primary body cells.

42. The preparation of claim 41, wherein the media are derived from culture media for primary fibroblasts and keratinocytes.

43. The preparation of claim 41, wherein the complete media are supplemented with serum substitutes.

44. The preparation of claim 27, wherein the preparation further comprises one or more skin culture media selected from one or more of DMEM/HAM's F-12 (1:1) and MCDB 153.

45. The preparation of claim 27, wherein the preparation further comprises a citrate buffer.

46. The preparation of claim 27, wherein the preparation is based on one of an aqueous gel, an O/W emulsion, a WIOIW emulsion, a W/O emulsion, a microemulsion and a cosmetic stick.

47. The preparation of claim 27, wherein the preparation further comprises at least one of Q10, AGR, Zn orotate, carnitine, creatine and taurine.

48. An article which comprises the preparation of claim 27, wherein the article is selected from a wound covering, a skin covering, a patch, a pad, a tissue and a bandage.

49. The preparation of claim 27, wherein the preparation is present as a constituent of a polyurethane matrix.

50. The preparation of claim 27, wherein the preparation is present as one of an aqueous surfactant preparation, an emulsion, an ointment, a cream, a gel, a dusting powder, a mask, a matrix bandage, a gel bandage, a foam and an aerosol preparation.

51. A skin care method, wherein the method comprises applying to skin the preparation of claim 27.

52. A skin treatment method, wherein the method comprises applying to injured or diseased skin the preparation of claim 27.

53. A method of wound management or wound healing, wherein the method comprises applying to a wound the preparation of claim 27.

54. The method of claim 53, wherein the preparation is comprised in a wound covering.

55. The method of claim 54, wherein the wound covering comprises a polyurethane.

56. A method of generating a complete partial skin, wherein the method comprises applying the preparation of claim 27 to a surface of a body from which skin has been removed.

57. A method of preventing or reducing scar tissue, wherein the method comprises applying to a wound the preparation of claim 27.

58. A process for preparing a cosmetic or dermatological preparation comprising at least one of collagen and a derivative thereof; at least one of chitosan and a derivative thereof, having a degree of acetylation of up to 50%; and at least one of a glycosaminoglycan and a derivative thereof;

wherein the process comprises

(a) providing a solution comprising water and at least one of collagen and a derivative thereof;

(b) adding at least one of chitosan and a derivative thereof, having a degree of acetylation of up to 50%, to the solution of (a) to form a mixture; and

(c) adding at least one of a glycosaminoglycan and a derivative thereof to the mixture of (b) to form a further mixture.

59. The process of claim 58, wherein the solution of (a) further comprises a cell culture medium.

60. The process of claim 58, wherein the glycosaminoglycan comprises at least one of chondroitin 4-sulfate and chondroitin 6-sulfate.

61. The process of claim 58, wherein the process further comprises (d) lyophilizing the mixture of (c) to provide an aerogel.

62. The process of claim 61, wherein the process further comprises converting the aerogel of (d) into a hydrogel by introducing the aerogel into one of an aqueous phase, an active ingredient phase and a culture media phase of the cosmetic or dermatological preparation.

63. The process of claim 61, wherein the process further comprises processing the aerogel of (d) into a polyurethane matrix.