WO2001091821A1 - Supports a base de collagene pour l'ingenierie tissulaire et la preparation de biomateriaux - Google Patents
Supports a base de collagene pour l'ingenierie tissulaire et la preparation de biomateriaux Download PDFInfo
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- WO2001091821A1 WO2001091821A1 PCT/FR2001/001631 FR0101631W WO0191821A1 WO 2001091821 A1 WO2001091821 A1 WO 2001091821A1 FR 0101631 W FR0101631 W FR 0101631W WO 0191821 A1 WO0191821 A1 WO 0191821A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0697—Artificial constructs associating cells of different lineages, e.g. tissue equivalents
- C12N5/0698—Skin equivalents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/60—Materials for use in artificial skin
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0068—General culture methods using substrates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/09—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells
- C12N2502/094—Coculture with; Conditioned medium produced by epidermal cells, skin cells, oral mucosa cells keratinocytes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
- C12N2502/1323—Adult fibroblasts
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/04—Screening or testing on artificial tissues
- C12N2503/06—Screening or testing on artificial skin
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/54—Collagen; Gelatin
Definitions
- collagen has proven to be an irreplaceable substrate for the production of artificial tissues containing living cells.
- biomaterials obtained are increasingly used in the pharmaceutical field and their future seems very promising for the preparation of injured connective tissues or for gene therapy by allowing the introduction and survival of modified cells in a living organism. 10
- cosmetic and dermopharmaceutical industries are making increasing use of reconstructed skin, all the more so since in these disciplines, animal tests are used less and less .
- Document WO 99/19005 discloses a multilayer membrane comprising a predominantly collagen II matrix layer having a sponge texture covered on at least one side and preferably on both sides, with at least one barrier layer having a closed texture,
- the barrier layer is formed by a natural animal membrane (see page 7, lines 23 to 32 and page 8, lines 10 to 30).
- this barrier layer is to prevent the penetration and therefore the growth of native tissue cells because this membrane is dedicated to
- cartilage cells or chondrocytes have a much slower multiplication or regeneration rate than the regeneration rate of soft tissue cells such as fibroblasts and, therefore, it is necessary to isolate them for allow them to grow by avoiding being invaded by fast-growing soft tissue cells.
- This document leads to this solution by using a cell-tight barrier layer which protects the growth of collagen II cells which promote the growth of chondrocytes (see page 2, lines 15 to 20).
- a bilayer material is initially prepared, each layer of which is capable of allowing the growth of human living cells, which constitutes an unobvious and totally unexpected solution in relation to the state of the technical.
- Document WO 96/08277 relates to the use of a collagen membrane as a peritoneal regeneration prosthesis, constituting an earlier invention of the same applicant.
- a preferred embodiment is a mixed membrane comprising a collagen sponge on which a collagen gel has been bonded, the membrane being obtained by drying the collagen gel in a non-toxic gaseous fluid, see in particular the Example II, pages 12 and 13 of this PCT application.
- Example II it appears that the sponge obtained is compressed for 15 seconds under a pressure of 150 bars and that the mixed membrane is formed by pouring a 1% collagen gel onto this compressed sponge, this gel being then dried in the open air.
- the mixed membrane obtained is in fact produced from two essentially compact layers, which constitutes a structure different from that which is the subject of the present invention. Furthermore, in the case of the present invention, it has been unexpectedly discovered that the claimed bilayer structure is compatible with a seeding of at least one layer with living human cells, allowing their conservation, as well as their multiplication.
- document FR 2 679 778 constitutes another still earlier invention by the same similar applicant.
- Document EP 0 686 402 also constitutes an earlier document by the same applicant relating to a postoperative anti-adhesion collagen membrane comprising two layers, a collagen-based support completely covered with a layer of gelatin, the mixture being in the lyophilized state.
- the gelatin layer has the critical aim of achieving a membrane bonding effect avoiding adhesions and the gelatin has rapid resorption by dissolution at 37 ° C. in the presence of cells.
- This document is different from the present invention in that it uses a dermis obtained by the elimination of a skin, covered with a mixture of human keratinocytes previously isolated by a conventional method, mixed with human melanocytes also previously isolated by a method. known and which is then subjected to a co-culture.
- This document does not provide for a compact layer as provided in the context of the present invention and which can be seeded with human living cells, critically combined with a porous sub-layer which is clearly different from a de-epidermalized dermis.
- the document WO 91/16010 describes equivalents of composite living skins consisting first of all in buying commercially a membrane of bovine collagen sponge which is inoculated with fibroblast cells (see page 8, third and fourth paragraphs).
- the sponge After incubation, the sponge is inverted and the upper surface is laminated with non-porous collagen which can be treated with pepsin (see page 8 last paragraph) which is generally bovine collagen. It is stated that the purpose of treatment with pepsin is to remove the telopeptides (page 9, first four lines).
- the pH of the collagen solution is adjusted to a neutral pH, which allows precipitation.
- the collagen forms a thin film layer on the sponge and the whole is cultured at 37 ° C for 60 minutes (page 9, last sentence of the first paragraph). Then, cultured keratinocytes are inoculated on the laminated layer and a new culture is carried out again at pH 7.2 and 35 ° C for 10 days.
- the bilayer structure is formed initially and is made up critically of a collagen sponge coated with a compact layer, this compact layer providing unexpected technical effects such as resulting from the following description.
- the main difficulties to overcome for the production of supports intended for obtaining living artificial tissues are as follows: good mechanical resistance, low sensitivity to temperatures around 37 ° C, biological properties favorable to cell development and metabolism, low susceptibility to vis-à-vis the enzymatic degradation and, finally, for certain applications and in particular reconstructed skin, preferable presence of a bilayer structure in which one of the layers is as compact as possible and the other porous.
- the object of the present invention is to solve these problems which remain unresolved both technically and industrially.
- the present invention makes it possible to solve all of these technical problems in a particularly simple, inexpensive manner, usable on an industrial scale and in particular on a cosmetic, dermopharmaceutical or pharmaceutical industrial scale.
- the present invention provides a new composite product forming a collagen support comprising at least one porous collagen layer coated on at least one face with an essentially compact collagen membrane produced either by a collagen film prepared by drying, preferably at air or in a gaseous fluid, a collagen gel, or by a very strongly compressed collagen sponge.
- the compression of the compressed collagen sponge is carried out at a pressure at least equal to approximately 50 bars, equivalent to approximately 50.10 5 Pascals (Pa), preferably between 50 bars (50.10 5 Pa) and 200 bars (200.10 5 Pa), possibly this compression taking place at a temperature between 20 and 80 degrees C °, even better between 40 C ° and 60 C °.
- a pressure at least equal to approximately 50 bars, equivalent to approximately 50.10 5 Pascals (Pa), preferably between 50 bars (50.10 5 Pa) and 200 bars (200.10 5 Pa), possibly this compression taking place at a temperature between 20 and 80 degrees C °, even better between 40 C ° and 60 C °.
- this composite product is characterized in that the abovementioned collagen product is chosen from collagen and a mixture of collagen with a polysaccharide, in particular a glycosaminoglycan, chitosan, and chitosan derivatives, cellulose and derivatives cellulose, dextran and its derivatives, an alginate, an alginate derivative, a carrageenan.
- a polysaccharide in particular a glycosaminoglycan, chitosan, and chitosan derivatives, cellulose and derivatives cellulose, dextran and its derivatives, an alginate, an alginate derivative, a carrageenan.
- this composite product is characterized in that at least one of the two layers, respectively the porous layer and the essentially compact membrane, comprises living cells, normal or genetically modified, or malignant in particular from young or old subjects.
- the living cells are chosen from the group consisting of fibroblasts, keratinocytes, melanocytes, langerhan cells of blood origin, endothelial cells of blood origin, blood cells, in particular macrophages or lymphocytes, adipocytes, sebocytes , bone cells, chondrocytes, osteoblasts, Merkel cells of blood origin, normal or genetically modified or malignant.
- the composite product is characterized in that it contains normal or genetically modified or malignant fibroblasts in the porous layer and normal or genetically modified or malignant living cells, on the surface of the compact membrane in particular chosen from keratinocytes, melanocytes, Merkel cells of blood origin, langerhan cells of blood origin, sebocytes, cells of blood origin, nerve cells.
- the essentially compact membrane is prepared prior to combination with the porous layer, preferably comprising a collagen sponge, in particular by preparing the membrane and depositing it on a gel collagen before the whole is frozen and lyophilized to obtain said composite product.
- the collagen sponge and / or the collagen film and / or the collagen membrane comprises collagen of mammalian origin, in particular of bovine origin.
- the collagen sponge and / or the collagen film and / or collagen membrane comprises collagen of marine origin, preferably derived from fish skins of the family.
- teleosts more particularly fish with areas of depigmented skin, even more particularly flat fish, even better those which are sinned industrially, such as for example sole, flounder, turbo, catfish whose non-ventral skins pigments can be easily separated by skinning.
- the fish skin preferred as a source of collagen extraction usable according to the present invention is that of sole.
- such a collagen sponge and / or a collagen film and / or a collagen membrane obtained from collagen of marine origin, preferably from the family of teleosts may be biocompatible with human living cells used for the manufacture of reconstructed skins and that these living human cells can not only remain alive but also be able to multiply.
- the collagen of mammalian origin, in particular of bovine origin, or preferably of marine origin, in particular of fish skins of the teleost family can be crosslinked either chemically, or by physical crosslinking as will be described later in the context of the manufacturing process. It is particularly unexpected that such crosslinking can be used in the context of the production of a biomaterial biocompatible with human cells which are used in the rest of the process for the manufacture of reconstructed skins.
- biocompatible is meant in the context of the present invention that the biomaterial is not toxic towards human living cells and that it also allows their growth or multiplication. It should be noted that the crosslinking generally has the effect of making the material non-biocompatible, therefore toxic to living cells and therefore cannot allow their growth.
- the composite product according to the invention is characterized in that at least one of the two layers is produced from a collagen gel containing a mixture of soluble collagen and insoluble collagen, for example in the form of fibers.
- the collagen can be type I and / or type III collagen.
- the present invention also covers a process for manufacturing a composite product comprising at least one porous collagen layer coated on at least one face with an essentially compact collagen membrane, characterized in that: a) everything is prepared first the essentially compact collagen membrane, either by drying a first collagen gel, preferably by air drying or using a gaseous fluid, or by compression of a collagen sponge obtained by freezing- lyophilization of a collagen gel; b) a second collagen gel is prepared separately; c) either the essentially compact membrane is deposited on the second collagen gel, or the second collagen gel is poured onto the essentially compact membrane; and d) finally freezing-lyophilization of the assembly to obtain said composite product.
- the compression step takes place at a temperature between 20 to 80 ° C, more preferably between 40 ° C and 60 ° C.
- this process is characterized in that one uses for the preparation of the collagen sponge and / or the collagen film and / or the collagen membrane, either collagen or a mixture of collagen with a polysaccharide, in particular a glycosaminoglycan, chitosan, chitosan derivatives, cellulose and cellulose derivatives, dextran and its derivatives, an alginate, an alginate derivative, a carrageenan.
- a polysaccharide in particular a glycosaminoglycan, chitosan, chitosan derivatives, cellulose and cellulose derivatives, dextran and its derivatives, an alginate, an alginate derivative, a carrageenan.
- the method is characterized in that a crosslinking of at least one of the two layers or of both is carried out.
- the aforementioned crosslinking is a physical crosslinking, in particular a thermal dehydration hot under vacuum or DHT, or a chemical crosslinking, in particular with diphenylphosphorylazide or DPPA, with an aldehyde such as glutaraldehyde, with carbodiimide, or succinimide.
- a compound promoting cell development in particular a growth factor, in particular a cytokine, a chemokine, is added during manufacture.
- the method according to the invention is characterized in that there is provided a step of introducing living cells, normal or genetically modified, or malignant in at least one of the two layers.
- said living cells are chosen from the group consisting of fibroblasts, keratinocytes, melanocytes, langerhan cells of blood origin, endothelial cells of blood origin, blood cells, in particular macrophages or lymphocytes, chondrocytes, bone cells, in particular osteoblasts, Merkel cells of blood origin, sebocytes, adipocytes, nerve cells.
- the method is characterized in that fibroblasts are introduced into the porous layer.
- the method is characterized in that living cells are deposited on the surface of the compact membrane, in particular chosen from keratinocytes, melanocytes, Merkel cells of origin blood, langerhans cells of blood origin, sebocytes, cells of blood origin, nerve cells.
- the method is characterized in that the living cells are brought either by culture sequentially, either by concomitant culture between the different types of cells, these cells coming from culture or biopsy.
- the present invention also covers the use of the composite product forming a collagen support as previously defined, or as obtained by the process as previously defined, or as resulting from the following description made in particular in relation with the examples for which any characteristic, which appears to be new compared to any prior art, is claimed as such in its function and in its generality, for the manufacture of artificial skins intended in particular for carrying out in vitro tests potentially active substance, or to reconstruct damaged skin areas in vivo.
- artificial skins can be obtained either substantially exclusively from young cells, or substantially exclusively from old cells, in particular for studying the process of tissue aging and in particular for the skin and possibly testing the effectiveness of principles. active on this process.
- reconstructed tissue capable of overcoming the deficiencies of damaged tissue: skin, cartilage, bone, tendons, corneas;
- any characteristic, which appears new compared to any prior art, is claimed in its function and in its generality, independently of the context of the example.
- Examples 6 to 13 constitute presently preferred embodiments of the composite products according to the invention forming a collagen support.
- Example 14 relates to comparative tests demonstrating the advantage of the composite products according to the invention as collagen support for the manufacture of artificial skins intended in particular for carrying out in vitro tests of effectiveness of potentially active substance, or for reconstructing in vivo damaged skin areas.
- the temperature is given in degrees Celsius
- the pressure is atmospheric pressure
- the percentages are given by weight unless otherwise indicated.
- Examples 16 to 24 constitute examples of preparation of collagen-based support of aquatic origin usable for the preparation of reconstructed skins
- Examples 25 to 27 relate to tests in particular comparative, using collagen of aquatic origin under the prepared forms in some of Examples 16 to 24, compared to collagen of bovine origin, allowing the manufacture of artificial skins.
- FIG. 1 shows a sectional view, after marking by conventional histological staining, of a composite product according to the present invention produced from a lower porous layer of bovine collagen, surfaced on the upper face of an upper membrane essentially compact collagen produced by a collagen film prepared by air drying a collagen gel, under the conditions of Example 6,
- FIG. 2 shows a similar section obtained with a simple porous matrix prepared with the same gel bovine collagen, but not surfaced, that is to say under the conditions of Example 1, showing the presence of inclusion of keratinocytes in depth, not limited to the surface,
- FIG. 4 shows the inductor effect of a fermented malt extract commercially available under the trade mark ® Basaline, COLETICA, France on the production of laminins in mature reconstructed skin, the amount of laminins produced also being expressed as percent of control ,
- FIG. 5 shows the same compensating effect of the fermented malt extract or Basaline ®, with the ordinate the amount of laminins produced as percent of control,
- FIG. 6 shows the proliferation of normal human fibroblasts in equivalent dermis, with the time expressed in days and on the abscissa and the optical density x 1000 on the ordinate with units increasing by 100;
- the curve with the diamonds is that obtained using a porous matrix of aquatic collagen, in this case fish, as support, and the curve with squares is obtained with bovine collagen;
- FIG. 7 represents a similar proliferation curve of fibroblasts in equivalent dermis with the time expressed in days and in abscissa.
- the curve with the solid diamonds represents the fluorescence obtained in the context of test 1; the curve with the square that obtained with test 2; the curve with the empty triangles being obtained with test 3 and finally the curve with the crosses being that obtained with test 4.
- a gel is prepared from previously washed calf skins
- the total duration of depilation is 36 hours.
- the skins are then stubborn in a bath containing ammonium chloride (3%) and sodium metabisulfite (0.5%), at the rate of 400 g of skin for 50 ml of bath.
- the total duration of this bath is 2 hours and thirty minutes.
- the salts are removed by two successive washes with water (15 minutes per wash), at the rate of 200 ml of water per 100 g of tissue.
- the ground material is then kneaded for one hour in order to obtain a paste.
- the gel is obtained by continuously passing the dough through an ultrasonic treatment device of the UTL T / -6 type.
- This gel has a concentration between 0.7 and 2% in collagen, the proportion of acid-soluble collagen varying from 10 to 20%, compared to insoluble collagen.
- the collagen lyophilisate is incubated for 24 h in a solution containing 5 to 250 ⁇ l of DPPA / g of collagen contained in 100 ml of dimethyl formamide (DMF).
- the collagen is then freed from DPPA by rinsing in 100 ml of DMF.
- the DMF is then eliminated by rinsing in 100 ml of a borate buffer solution pH 8.9 (sodium tetraborate 0.04 M, boric acid 0.04 M).
- the collagen is finally incubated overnight in the same borate buffer. Finally, the borate buffer is removed by rinsing with continuous permuted water for 6 h.
- NHS N-hydroxysuccinimide
- the collagen is crosslinked for 24 to 96 h in a solution containing 0.6 to 1% of GTA at 20 ° C. After rinsing with deionized water, the collagen is again lyophilized.
- Porous matrix prepared with the native collagen of Example 1 in combination with chitosan and a glycosaminoglycan as described in the European patent of May 29, 1991 No. 296078.
- Porous matrix described in Example 1 surfaced with a collagen film
- the collagen gel whose dry matter is between 0.3 and 0.8% is dried in an oven at 30 ° C or in a hood at the rate of 0.5 g of gel / cm of tray.
- the collagen gel it is possible to add 10 to 40% glycerol.
- the collagen dried under these conditions forms a transparent film.
- Porous matrix prepared with an acid-soluble collagen gel surfaced with a collagen film
- Example 6 The process is that indicated in Example 6, the only difference being the nature of the gel cast on the film which is made of acid-soluble collagen prepared according to a technique well known to those skilled in the art.
- EXAMPLE 8 Porous matrix prepared with an atelocollagen gel or surfaced with a collagen film
- Example 6 The process is that indicated in Example 6, the only difference being the nature of the gel cast on the film which is made of atelocollagen, that is to say collagen without telopeptide prepared according to a technique well known in the art. skilled in the art.
- Porous matrix consisting of collagen associated with chitosan and a glycosaminoglycan surfaced with a collagen film.
- the process is that indicated in Example 6, but in this case, the gel cast on the collagen film consists of collagen, chitosan, a glycosaminoglycan.
- the preparation of this gel is described in Example 5.
- EXAMPLE 10 All the porous matrices surfaced with a collagen film described previously can be crosslinked according to the techniques described in Examples 2, 3 and 4.
- EXAMPLE 11 Porous collagen alone matrix described in Example 1 surfaced with a compressed collagen sponge.
- the collagen gel prepared as in Example 1 and having a dry matter of between 0.3 and 1.5% is lyophilized so as to obtain a sponge having a weight of between 0.5 and 2 g / cm 2 .
- the lyophilisate is compressed for 5 to 60 seconds, at a temperature between 20 and 60 ° C and a pressure between 50 and 200 bars (50 to 200.10 5 Pa).
- Example 1 The collagen gel described in Example 1 is deposited at a rate of 0.5 g per cm 2 in a lyophilization tray. The compressed sponge is then deposited on this gel and the whole is freeze-dried. A porous collagen sponge surfaced with a compressed collagen sponge is thus obtained. The whole is crosslinked by DHT as described in Example 1.
- Porous matrix consisting of collagen, chitosan and glycosaminoglycan as described in Example 5 and surfaced with the compressed sponge.
- the collagen, chitosan and glycosaminoglycan gel prepared according to the method of Example 5, is deposited at a rate of 0.5 g per cm in a freeze-drying tray, then the compressed sponge is deposited on this gel and the assembly is lyophilized. The lyophilisate is then crosslinked by DHT as described in Example 1.
- Reconstructed skins prepared respectively either using the porous matrix crosslinked by DPPA described in example 2, or thanks to the porous matrix, crosslinked by DPPA, of example 2, surfaced by a compressed collagen sponge of which the whole is crosslinked by the DPPA, according to Example 13, in order to make a comparison between a composite product comprising a porous collagenous layer surfaced by an essentially compact membrane according to the invention and a product comprising a porous collagenous layer alone not surfaced.
- Normal human fibroblasts are used resulting from samples taken indifferently from elderly or young subjects, which are recovered and which are developed in a conventional manner for those skilled in the art to recover them between the sixth and the tenth passage. .
- Seeding is carried out at 250,000 cells per cm 2 of porous matrix, respectively either the comparison product comprising only the porous matrix crosslinked by the DPPA of Example 2, or the composite product according to the invention comprising the porous matrix crosslinked by the DPPA of example 2, surfaced by a compressed collagen sponge, the whole of which is crosslinked by the DPPA, according to example 13.
- the culture medium is composed of DMEM / HAM F12 50/50 (v / v) added to 10% by weight of fetal calf serum, 100 IU / ml of penicillin, 25 ⁇ g / ml of gentamycin, 1 ⁇ g / ml d amphotericin B, 50 ⁇ g / ml of vitamin C.
- a culture is carried out for three weeks by changing the medium three times a week.
- Seeding is carried out at 250,000 cells per cm 2 of surface, ie of the surface of the porous matrix crosslinked with the DPPA of Example 2; either of the composite product according to the invention comprising the porous matrix crosslinked by the DPPA of example 2, surfaced by a compressed collagen sponge, the whole of which is crosslinked by the DPPA, according to example 13 and, in this case, the keratinocytes are sown on the surface of the essentially compact collagen membrane.
- the culture of these products comprising both a seeding of fibroblasts and keratinocytes takes place in a Green medium composed of: DMEM supplemented with: 30% HAM F12, 10% fetal calf serum,
- EGF Epidermal Growth factor 10 ng / ml, commercially available insulin under the brand name UMUUfNE®
- This culture is carried out for a week by changing the media every day.
- step b) After having carried out the culture of step b) for one week by changing media every day, the surface layer containing the keratinocytes emerges at the air-liquid interface, while the layer containing the fibroblasts remains immersed, followed a culture for three weeks in emersion medium composed of:
- DMEM fetal calf serum
- penicillin 100 IU / ml of penicillin
- a reconstructed skin is obtained composed of a reconstructed dermis, the fibroblasts having colonized the three-dimensional collagen matrix, covered by a multi-layered epidermis.
- the surfacing of porous matrices with an essentially compact layer to obtain a composite product according to the invention makes it possible to obtain, after three weeks of preparation of the equivalent dermis, a greater quantity of fibroblasts on the surface of the collagenic matrices before epidermization. .
- the procedure is essentially as described in example 14 concerning the cultures using the same composite product according to the present invention comprising a porous collagenous layer or matrix crosslinked by the DPPA described in example 2, surfaced by a sponge of compressed collagen, the whole of which is crosslinked by the DPPA, according to Example 13, by proceeding as follows:
- step a the porous matrices of the composite product of the invention were firstly seeded with normal human dermal fibroblasts from either pools of young cells or pools of cells mature or aged, and a culture is carried out for 21 days under the conditions described in Example 14 step a.
- epidermal sheets prepared separately from keratinocytes originating either from pools of young cells or from pools of mature cells are seeded on the surface of the essentially compact collagen membrane of the composite product.
- a culture is carried out for 14 days under the conditions described in Example 14b.
- FIG. 3 shows that the reconstructed mature skins contain approximately half as much laminins as the reconstructed young skins (PRJ) serving as a 100% control.
- COLETICA on the production of laminins in young and mature reconstructed skin
- the laminins contained in the incubation media were quantified by ELISA.
- the tension at 100% is made up of the level of laminins in Aged Reconstructed Skin or PRA.
- Figure 4 shows that the active ingredient extracted from fermented malt, or BASALINE ® , was capable of stimulating the production of laminins in mature reconstructed skins. Under the same conditions, the active extract of fermented malt or Basaline ® does not alter how significant the production of laminin in skins youth rebuilt, shown in Figure 5. It is found that the active ingredient or fermented malt extract
- BASALINE ® increases by 65% the production of laminins from mature reconstructed skin.
- this active principle does not affect the physiological processes involved in regulating the production of laminins from young reconstructed skin.
- FIG. 5 shows that the difference between the production of laminins from young reconstituted skins and from mature reconstituted skins can be reduced by 65% by the active ingredient, used to 0.5%.
- a collagen gel is prepared from ground ventral sole skins then washed with a phosphate buffer pH 7.8, the composition of which is: 0.78 g / 1 of potassium dihydrogen phosphate and 21.7 g / 1 of disodium monohydrogen phosphate.
- the washing is carried out with stirring for one hour at a rate of 5 l of buffer for 1 kg of ground material.
- the phosphate is then removed by two successive washes with deionized water, then by continuous centrifugation at 4000 rpm (Rousselet decanter), at the rate of 5 l of water per 1 kg of ground material.
- the ground product is then acidified with a 0.25 M acetic acid solution at a rate of 1 kg of ground material for 10 1 of solution.
- the gel is then centrifuged at 4000 rpm for 5 minutes.
- the gel which will be used consists of the supernatant obtained whose collagen concentration is between 0.5 and 2%.
- This gel is poured into a lyophilization tray at a rate of 20 g / cm. It is then lyophilized after freezing at -30 ° C and heating to + 32 ° C. The lyophilization lasts a total of 16 hours under a pressure of 400 microbars.
- the matrix obtained is then crosslinked by hydrothermal dehydration (DHT). This consists of heating in an oven at 110 ° C under a vacuum of 400 microbars for 16 hours.
- DHT hydrothermal dehydration
- the collagen matrix of Example 16 is incubated for 24 h in a solution containing 5 to 250 ⁇ l of DPPA / g of collagen contained in 100 ml of dimethylformamide (DMF).
- the collagen is then freed from DPPA by rinsing in 100 ml of DMF.
- the DMF is then eliminated by rinsing in 100 ml of a borate buffer solution pH 8.9 (sodium tetraborate 0.04 M, boric acid 0.04 M).
- the collagen is finally incubated overnight in the same borate buffer.
- the borate buffer is removed by rinsing with continuous permuted water for 6 h.
- the aquatic collagen matrix of Example 16 is crosslinked with EDC (Ethyldimethylaminopropyl carbodiimide) at the concentration of 0.23 to 0.69 g / g of collagen, and with NHS (N-hydroxysuccinimide) at the concentration of 0.42 g / g of collagen.
- EDC Ethyldimethylaminopropyl carbodiimide
- NHS N-hydroxysuccinimide
- the collagen After rinsing with deionized water, the collagen is again lyophilized.
- the porous aquatic collagen matrix of Example 16 is crosslinked for 24 to 96 h in a solution containing 0.6 to 1% of GTA at 20 ° C. After rinsing with deionized water, the collagen is again lyophilized.
- Porous matrix prepared with the native aquatic collagen of Example 16 in combination with chitosan and a glycosaminoglycan as described in the European patent of May 29, 1991 No. 296078
- Porous matrix described in Example 16 surfaced with a collagen film
- the collagen gel the dry matter of which is between 0.3 and 0.8%, is dried in an oven at 30 ° C or in a hood at the rate of 0.5 g of gel / cm 2 of tray.
- the native aquatic collagen gel of 0.5% to 2% in dry matter is deposited at the rate of 0.5 g per cm 2 in a lyophilization tray, then the collagen film is deposited on this gel and the whole is lyophilized.
- the lyophilisate obtained is crosslinked with DHT.
- EXAMPLE 22 Porous collagen alone matrix described in Example 16 surfaced with a compressed collagen sponge.
- the collagen gel prepared as in Example 16 and having a dry matter of between 0.3 and 1.5% is lyophilized so as to obtain a sponge having a weight of between 0.5 and 2 g / cm 2 .
- the lyophilisate is compressed for 5 to 60 seconds, at a temperature between 20 and 60 ° C and a pressure between 50 and 200 bars (50 to 200.10 5 Pa).
- Example 16 The collagen gel described in Example 16 is deposited at a rate of 0.5 g per cm in a freeze-drying tray. The compressed sponge is then deposited on this gel and the whole is freeze-dried. A porous sponge from collagen surfaced with a compressed collagen sponge is thus obtained. The whole is crosslinked by DHT as described in Example 16.
- EXAMPLE 23 Porous matrix consisting of collagen, chitosan and glycosaminoglycan as described in Example 20 and surfaced with the compressed sponge.
- the collagen, chitosan and glycosaminoglycan gel prepared according to the method of Example 20, is deposited at a rate of 0.5 g per cm 2 in a lyophilization tray, then the compressed sponge is deposited on this gel and the whole is freeze-dried. The lyophilisate is then crosslinked by DHT as described in Example 16.
- Example 17 an aquatic porous matrix crosslinked with DPPA is produced on the one hand, according to Example 17.
- a fabricates a comparative porous matrix with collagen of bovine origin, also called bovine matrix, crosslinked with DPPA, under the same conditions as those of Example 17.
- the culture of these aquatic and bovine matrices is carried out respectively in medium composed of DMEM / HAM F 12 in a 50/50 ratio (v / v) supplemented with 10% fetal calf serum, 100 IU / ml of penicillin, 25 ⁇ g / ml of gentamycin, 1 ⁇ g / ml of amphotericin B, 50 ⁇ g / ml of vitamin C.
- This culture is carried out for 1 month by changing the culture medium 3 times a week.
- MTT ie 3- (4- (dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide
- the optical density of the transformation product (formazan blue) is read at 550 nm after self-dissolution in DMSO.
- the optical density obtained is proportional to the activity of the succinate dehydrogenases which are capable of carrying out the transformation of the light yellow tetrazolium MTT salt into blue crystals of formazan.
- the curve with the diamonds is that obtained with the aquatic matrix and the curve with the squares is that obtained with the bovine matrix. It is observed from the results that in a completely su ⁇ renant manner, the aquatic matrix not only constitutes a support allowing the survival of normal human fibroblasts but also the cellular proliferation of these normal human fibroblasts, while even constituting a much better culture support during first three weeks.
- the aquatic collagen is, in a su ⁇ renant manner, particularly suitable for producing a tissue engineering support in particular for applications in vitro and even and especially in vivo to constitute biomaterials containing living cells and in particular and preferably living cells of human beings.
- the assay is carried out by the Pierce microBCA method.
- the cell density was evaluated by an MTT test under the conditions described above.
- the relative protein content corresponds to the protein content reduced to 1 unit of cell density expressed in optical density or OD, in order to reason at equivalent cell concentration.
- the results obtained are listed in Table II below:
- fibroblasts could be observed in matrices of bovine and aquatic origin. In both types of matrices, we note the presence of an abundant neosynthesized extracellular matrix. The neosynthesized extracellular matrix can be differentiated by the periodic striation of the deposited collagen fibers compared to the collagen clusters forming the three-dimensional matrix of the starting sponge.
- Test 1 For this test, a porous matrix is produced in the form of a porous sponge from an aquatic collagen gel prepared from 1.3% by weight of aquatic collagen, which is frozen at ⁇ 80. ° C and which is subjected to a standard lyophilization in accordance with Example 17 and which is then subjected to crosslinking with DPPA in a proportion of 250 ⁇ l / g of sponge in the dry state.
- a porous support in the form of an aquatic sponge is prepared from an aquatic collagen gel comprising 0.7% by weight of aquatic collagen, which is subjected to freezing at -80 ° C., then to standard lyophilization and crosslinking with DPPA at 250 ⁇ l / g dry as in test 1.
- a porous support comprising an aquatic collagen sponge obtained from an aquatic collagen gel comprising 1.1% by weight of aquatic collagen, which is subjected to freezing at -80 ° C., then to standard lyophilization and crosslinking with DPPA at 250 ⁇ l / g dry as in test 2, the difference residing in a proportion of 1.1% by weight of aquatic collagen.
- the aquatic collagen is derived, as in Example 17, from the ventral sole skin.
- Example 25 Is used as in Example 25, normal human fibroblasts but which are taken at 8 th passage. Seeding is carried out at the rate of 275,000 cells per cm.
- the culture medium is composed of DMEM / HAM F12 50/50 (v / v) added with 10% by weight of fetal calf serum, 100 IU / ml of penicillin, 25 ⁇ g / ml of gentamycin, 1 ⁇ g / ml of amphotericin B, 50 ⁇ g / ml of vitamin C.
- the culture is carried out for 1 month by changing the medium 3 times a week.
- Alamar Blue is added at a rate of 2% by weight of a culture medium used, when it is desired to measure the cell viability on a sample taken from the culture medium. • After incubation at 2 h 20 at 37 ° C., the fluorescence is read, on the basis of an excitation at 530 nm and an emission at 590 nm).
- the intensity of the fluorescence obtained is proportional to the metabolic activity of the cells.
- the cell viability measurement is carried out on 10 samples after 1, 4 6, 11 17 days of culture.
- the results of the table UI are also the subject of attached figure 7. They show the fibrobiastic proliferation curves in equivalent dermis.
- the solid diamond curve is that performed with test 1; the curve in full square is that carried out with test 2; the triangle curve is that carried out with test 3 and the curve with the crosses is that carried out with test 4.
- the various matrices prepared can allow good fibrobiastic growth after 17 days of culture.
- the fibroblasts adhere well to their three-dimensional support and divide very quickly in order to colonize the matrix.
- the proliferation profile is very slightly variable from one type of matrix to another, but after 17 days of culture, the fibrobiastic density is comparable whatever the manufacturing process.
- This test is similar to that of Example 25, except that a histology with immunostaining is carried out.
- Example 25 These are equivalent dermes from Example 25, the culture being carried out under the conditions of Example 25.
- This culture is therefore carried out for three weeks by changing the medium three times a week, the seeding of normal human fibroblasts having taken place at 300,000 cells per cm 2 as was indicated in Example 25.
- the fixation is carried out with paraformaldehyde at a content of 4% by weight, then a dehydration and a paraffin inclusion are carried out.
- Haidenhain after dewaxing and rehydration Haidenhain after dewaxing and rehydration.
- Tissue Tek OCT compound is carried out, that is to say an inclusion liquid supplied by Miles, Elkhart, Indiana, USA, and a cold cut at 7 ⁇ m. Immunostaining is performed as follows: i. With a first anti-human collagen type I antico ⁇ s of rabbit (dilution),
- the supports constituted respectively by an aquatic matrix and a bovine matrix form more or less loose pores in which the fibroblasts adhere.
- On the surface there is a higher proportion of fibroblasts forming a favorable surface treatment of the equivalent dermis for producing reconstructed skin.
- the distribution of fibroblasts is homogeneous in aquatic and bovine sponges.
- the matrix of aquatic origin is only very weakly marked by the antico ⁇ s anti-human collagen.
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DE10196234T DE10196234B4 (de) | 2000-05-26 | 2001-05-25 | Verfahren zur Herstellung eines Verbundprodukts aus Kollagen für die Gewebebildung und seine Verwendung |
JP2001587833A JP3865635B2 (ja) | 2000-05-26 | 2001-05-25 | 組織工学及び生体材料製造のためのコラーゲンベースによる支持体 |
KR20027015963A KR100520944B1 (ko) | 2000-05-26 | 2001-05-25 | 인공조직 및 생체재료 제조용 콜라겐 지지체 |
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US09/616,526 US6790454B1 (en) | 2000-05-26 | 2000-07-14 | Processes for the preparation of novel collagen-based supports for tissue engineering, and biomaterials obtained |
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1989008467A1 (fr) * | 1988-03-15 | 1989-09-21 | Imedex | Patch de chirurgie viscerale |
WO1991016010A1 (fr) * | 1990-04-24 | 1991-10-31 | Mark Eisenberg | Equivalents composites de peau vivante |
FR2678624A1 (fr) * | 1991-07-04 | 1993-01-08 | Coletica | Utilisation de la peau non pigmentee de poisson, en particulier de poisson plat comme nouvelle source industrielle de collagene, procede d'extraction, collagene et biomateriau ainsi obtenus. |
FR2679778A1 (fr) * | 1991-08-02 | 1993-02-05 | Coletica | Utilisation de collagene reticule par un agent de reticulation pour la fabrication d'une membrane suturable, biocompatible, a resorption lente, ainsi qu'une telle membrane. |
EP0686402A1 (fr) * | 1994-06-08 | 1995-12-13 | Coletica | Membrane collagénique anti-adhérence post-opératoire |
WO1996008277A1 (fr) * | 1994-09-15 | 1996-03-21 | Coletica | Utilisation de membranes collageniques comme protheses de regeneration peritoneale |
EP0789074A1 (fr) * | 1996-01-23 | 1997-08-13 | L'oreal | Equivalent de peau comprenant des cellules de Langerhans |
WO1999019005A1 (fr) * | 1997-10-10 | 1999-04-22 | Ed Geistlich Söhne Ag Für Chemische Industrie | Membrane utilisee dans la regeneration tissulaire |
FR2783429A1 (fr) * | 1998-09-18 | 2000-03-24 | Imedex Biomateriaux | Materiau collagenique bicomposite,son procede d'obtention et ses applications therapeutiques |
-
2001
- 2001-05-25 WO PCT/FR2001/001631 patent/WO2001091821A1/fr active IP Right Grant
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1989008467A1 (fr) * | 1988-03-15 | 1989-09-21 | Imedex | Patch de chirurgie viscerale |
WO1991016010A1 (fr) * | 1990-04-24 | 1991-10-31 | Mark Eisenberg | Equivalents composites de peau vivante |
FR2678624A1 (fr) * | 1991-07-04 | 1993-01-08 | Coletica | Utilisation de la peau non pigmentee de poisson, en particulier de poisson plat comme nouvelle source industrielle de collagene, procede d'extraction, collagene et biomateriau ainsi obtenus. |
FR2679778A1 (fr) * | 1991-08-02 | 1993-02-05 | Coletica | Utilisation de collagene reticule par un agent de reticulation pour la fabrication d'une membrane suturable, biocompatible, a resorption lente, ainsi qu'une telle membrane. |
EP0686402A1 (fr) * | 1994-06-08 | 1995-12-13 | Coletica | Membrane collagénique anti-adhérence post-opératoire |
WO1996008277A1 (fr) * | 1994-09-15 | 1996-03-21 | Coletica | Utilisation de membranes collageniques comme protheses de regeneration peritoneale |
EP0789074A1 (fr) * | 1996-01-23 | 1997-08-13 | L'oreal | Equivalent de peau comprenant des cellules de Langerhans |
WO1999019005A1 (fr) * | 1997-10-10 | 1999-04-22 | Ed Geistlich Söhne Ag Für Chemische Industrie | Membrane utilisee dans la regeneration tissulaire |
FR2783429A1 (fr) * | 1998-09-18 | 2000-03-24 | Imedex Biomateriaux | Materiau collagenique bicomposite,son procede d'obtention et ses applications therapeutiques |
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