WO2011051983A1 - Fibre de tissu animal modifiée biologiquement in vitro et son utilisation dans l'industrie textile - Google Patents

Fibre de tissu animal modifiée biologiquement in vitro et son utilisation dans l'industrie textile Download PDF

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Publication number
WO2011051983A1
WO2011051983A1 PCT/IT2009/000488 IT2009000488W WO2011051983A1 WO 2011051983 A1 WO2011051983 A1 WO 2011051983A1 IT 2009000488 W IT2009000488 W IT 2009000488W WO 2011051983 A1 WO2011051983 A1 WO 2011051983A1
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Prior art keywords
microbeads
porous
fiber
bioengineered
cells
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English (en)
Inventor
Paolo Netti
Giorgia Imparato
Francesco Urciuolo
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DMD SOLOFRA SpA
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DMD SOLOFRA SpA
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Priority to PCT/IT2009/000488 priority Critical patent/WO2011051983A1/fr
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Classifications

    • 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
    • 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
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/005—Ingredients of undetermined constitution or reaction products thereof
    • 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
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
    • D01D5/00—Formation of filaments, threads, or the like
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F4/00—Monocomponent artificial filaments or the like of proteins; Manufacture thereof
    • D—TEXTILES; PAPER
    • D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
    • D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
    • D01F9/00—Artificial filaments or the like of other substances; Manufacture thereof; Apparatus specially adapted for the manufacture of carbon filaments

Definitions

  • the present invention relates to a bioengineered fiber of tissue of animal origin and to a process for its preparation by means of in vitro cell culture.
  • Such fiber may particularly, but not exclusively, be used as yarn for textile industry applications.
  • the new features requested for the textile products may be obtained by means of new production processes and/or by means of novel fibers.
  • the main advantage of the artificial fibers lies in that they can be developed for the specific applications that they will have to perform.
  • the woven fibers both natural and synthetic, are their use in the textile market. Because of the availability of materials with a wide spectrum of functionalities and technological features, the fibers are used in several fields such as sport, clothing, furnishing, medical-surgical, automotive, and aerospace.
  • leather is also currently used.
  • Leather is obtained from natural animal pelts after a series of complex tanning treatments, giving the final product properties of preservability, softness, mechanical strength, flexibility and color. These properties allow leather to be useful in a large number of industrial applications.
  • tanning processes have some important technological limitations, some of the main ones being the following:
  • animal cells dissociated from their original tissue are capable of creating new tissues when cultured in vitro onto three-dimensional supports in an optimal environment. It has been shown to be possible, for example, to form tubular structures from dissociated endothelial cells (Montesano et al., J. Cell. Biol. 97:1648, 1983) or cartilage tissue from isolated chondrocytes (Kisiday et al., Proc. Natl. Acad. Sci USA. 99:9996, 2002).
  • the cells are capable of reconstructing their typical tissue because they retain in vitro also their ability to produce extracellular matrix (ECM) (Webb et al, Biomaterials. 24:4681, 2003).
  • ECM extracellular matrix
  • the present invention aims to produce a bioengineered tissue yarn by using a new process in which particulate micro-carriers are seeded with eukaryote cells and the seeded micro-carriers are formed into a shaped assembly. By arranging in close mutual proximity under suitable culture conditions the cell seeded microcarriers are able to form a three-dimensional tissue equivalent due to their biological union. By employing appropriate techniques, the present invention aims to obtain a bioengineered tissue fiber suitable for. textile industry.
  • the present invention provides a process for preparing a bioengineered fiber suitable for the textile industry which does not require the production of a thin sheet of biological material to be subsequently processed into a shaped article.
  • the present invention aims at producing a bioengineered fiber-shaped product that can be used in the textile industry.
  • a process for preparing a bioengineered fiber of tissue of animal origin comprises the steps of:
  • the aim and objects of the invention are also achieved by a bioengineered fiber of tissue of animal origin obtainable by means of said process.
  • the aim and objects of the invention are also achieved by the use of said bioengineered fiber of tissue of animal origin in the textile industry, in particular as textile yarn.
  • Figure 1 shows the steps of a preferred embodiment of the process of the invention.
  • figure la- Id show the preparation of macroporous gelatin microbeads (a- gelatin-toluene emulsion; b- saturated toluene droplets in gelatin solution; c- formation of gelatin microbeads containing droplets of toluene; d- porous gelatin microbeads obtained after toluene washing with ethanol); figure le shown the dynamic fibroblast seeding on the gelatin porous microbeads;
  • figure If shows the microbeads covered with a tissue layer (microtissue); figure lg shows the injection and molding step of the microtissues in the porous microtube;
  • figure lh shows the tissue growth into the microtube
  • figure lhl shows the cell alignment and tissue deposition along tube axis
  • figure lh2 shows the synchronous microbeads degradation and neotissue formation
  • figure 2a shows a Scanning Electron Microscope (SEM) picture of a biodegradable porous gelatin microbead (bar is 10 ⁇ );
  • figure 2b shows a Scanning Electron Microscope (SEM) picture of a biodegradable porous gelatin microbead (bar is 20 ⁇ );
  • figure 3 shows a graph of the concentration of cells in the culture medium in function of time as a result of dynamic seeding
  • figure 4a shows an optical microscope picture of a microbead colonized by fibroblasts 2 hours after seeding and subjected to MTT assay
  • figure 4b shows an optical microscope picture of a microbead colonized by fibroblasts 5 hours after seeding and subjected to MTT assay
  • figure 4c shows an optical microscope picture of a microbead colonized by fibroblasts 24 hours after seeding and subjected to MTT assay
  • figure 4d shows an optical microscope picture of a microbead colonized by fibroblasts 48 hours after seeding and subjected to MTT assay
  • figure 4e shows an optical microscope picture of a microbead colonized by fibroblasts 96 hours after seeding and subjected to MTT assay
  • figure 5 a shows a picture of a hematoxylin/eosin stained 20% crosslinked microbead after 4 days of culturing of the fibroblasts seeded therein;
  • figure 5b shows a picture of a hematoxylin/eosin stained 10% crosslinked microbead after 4 days of culturing of the fibroblasts seeded therein;
  • figure 6a shows a SEM picture of a microbead after 4 days of culture of the fibroblasts seeded therein (bar is 100 ⁇ );
  • figure 6b shows an enlargement of the SEM picture of a microbead after 4 days of culture of the fibroblasts seeded therein (bar is 10 ⁇ );
  • figure 7a shows a SEM picture of the diameter of the PTFE microtube
  • figure 7b shows a SEM picture of the porosity of the PTFE microtube
  • figure 8 shows a scheme of the apparatus for injecting the seeded microbeads into the microtube
  • figure 9 shows a picture of a bioengineered fiber of tissue of animal origin obtained after 1 week of culture of the microbeads injected into the microtube;
  • figure 10a shows a picture of a hematoxylin/eosin stained fiber obtained after 3 weeks of culturing of the 20% crosslinked microbeads within the microtube;
  • figure 10b shows an enlargement of the picture of a hematoxylin/eosin stained fiber obtained after 3 weeks of culturing of the 20% crosslinked microbeads within the microtube;
  • figure 11a shows a picture of a hematoxylin/eosin stained fiber obtained after 3 weeks of culturing of the 10% crosslinked microbeads within the microtube;
  • figure l ib shows an enlargement of the picture of a hematoxylin/eosin stained fiber obtained after 3 weeks of culturing of the 10% crosslinked microbeads within the microtube;
  • figure 12a shows a picture of the ultrastructural analysis of the fiber obtained after 3 weeks of culture (bar is 100 nm).
  • figure 12b shows a picture of the ultrastructural analysis of the fiber obtained after 3 weeks of culture (bar is 500 nm).
  • bioengineered fiber refers to a fiber made by biological tissue obtained by means of in vitro culturing of eukaryotic cells, particularly fibroblasts, of animal origin.
  • microtissue refers to a complex consisting of a microcarrier surrounded by eukaryotic cells and a micrometric layer of their extracellular matrix.
  • the cells can be fibroblast of animal origin and the microcarriers can be porous microbeads.
  • the process herein described comprises several steps suitable to produce a bioengineered fiber.
  • the first step consists in seeding eukaryotic cells on biodegradable porous microbeads with a controlled and tunable degradation rate under dynamic seeding conditions, thereby obtaining microtissues.
  • the cells and the microbeads are maintained in condition of intermittent stirring followed by continuous stirring.
  • the dynamic cell seeding may be carried out by subjecting the cells and microbeads in a suspension to stirring phases having a duration time ranging from 3 to 15 minutes, preferably 5 minutes, alternated with rest phases having a duration time ranging from 15 to 60 minutes, preferably 30 minutes.
  • the alternating phases may be carried out for a total time of 5 to 12 hours, preferably 6 hours, at the end of which the cells and microbeads may be maintained under continuous stirring for a time ranging from 72 to 160 hours, preferably 90 hours.
  • the stirring rate may be set in the range of 20-40 rpm, more preferably 30 rpm, in order to maintain the microbeads and cells in suspensions while avoiding the damage induced by mechanical stress due to the stirring.
  • the dynamic cell seeding allows a better distribution of the cells onto the surface of the microbeads, creates an homogeneous culturing milieu, avoids microbeads aggregation and improves gas and nutrient exchange between the cell seeded microbeads and the culture medium.
  • the eukaryotic cells are fibroblasts, more preferably the fibroblasts are primary fibroblasts.
  • the fibroblasts may be bovine fibroblasts.
  • Fibroblasts seeding may be carried out in MEM Eagle-Earle BSS culture medium enriched with a solution of essential and non-essential amino acids (2x), 20% Fetal Bovine Serum (FBS) and glutamine 2mM.
  • the fibroblasts are trypsinised and 5xl0 5 cells per mg of microbeads are seeded on the microbeads according to the procedure herein described.
  • microbeads used in the process herein described act as a support for cell adhesion and further offer to the cells a protection against mechanically induced damage.
  • the process herein described may further comprise a step of preparing the porous microbeads prepared having controlled and tunable degradation rate before the step (a).
  • the microbeads may be for example gelatin microbeads with a controlled degradation rate, obtained by means of the "oil-in water-in oil" (O/W/O) double emulsion technique, by mixing an aqueous solution of gelatin containing a first surface-active agent, with a hydrophobic organic solvent containing a second surface-active agent.
  • O/W/O oil-in water-in oil
  • the O/W/O emulsion consists of a three-phase system wherein a water phase, containing a first hydrophobic phase, is dispersed in a second hydrophobic phase.
  • the O/W/O emulsion is prepared by mixing an aqueous solution of gelatin containing a first surface-active agent, such as a polyethoxylated sorbitol ester, with an hydrophobic organic solvent, for example toluene, containing a second surface-active agent, for example a sorbitol ester.
  • the first surface-active agent may be TWEEN 85 (sorbitan trioleate ethoxylated with 20 moles of ethylene oxide).
  • the second surface-active agent may be SPAN 85 (sorbitan trioleate).
  • SPAN 85 sorbitan trioleate
  • Addition of the hydrophobic organic solvent to the aqueous phase leads to formation of a hydrophobic phase in the form of organic solvent droplets trapped within the aqueous phase.
  • the organic solvent leads to saturation of the aqueous solution, the latter becomes dispersed in an external hydrophobic phase of the solvent. Gelatin microbeads with entrapped organic solvents droplets are thus formed.
  • the microbeads are washed with a solvent miscible with the hydrophobic solvent used for the preparation of the double emulsion which is not capable of dissolving gelatin, for example ethanol may be used when toluene is used in the emulsion. Accordingly, the droplets of hydrophobic organic solvent trapped within the microbeads are removed, thereby forming an extended network of micropores in the microbeads.
  • a solvent miscible with the hydrophobic solvent used for the preparation of the double emulsion which is not capable of dissolving gelatin for example ethanol may be used when toluene is used in the emulsion. Accordingly, the droplets of hydrophobic organic solvent trapped within the microbeads are removed, thereby forming an extended network of micropores in the microbeads.
  • the biodegradable porous microbeads may have a diameter ranging from 75 ⁇ to 150 ⁇ , more preferably 150 ⁇ .
  • the gelatin porous microbeads may be stabilized by means of a treatment with a cross linking agent, whose amount influences the rate of degradation of the microbeads. It is thus possible to modulate the rate of degradation of the microbeads by varying the concentration of the crosslinking agent during the stabilization, thereby synchronizing the rate of degradation of the gelatin microbeads with the rate of extracellular matrix synthesis of the cells seeded within the microbeads.
  • the cross linking agent which may be for example glyceraldehyde, may be used at concentration ranging from 10% w/w to 20% w/w of the microbeads weight.
  • the gelatine porous microbeads may be subjected to sterilization, so as to provide for a sterilized environment for the cells which will be seeded into the microbeads.
  • sterilization may be carried out by means of autoclaving or gamma-ray irradiation.
  • sterilization may be carried out in an autoclave at a temperature of 121°C for 15 minutes.
  • the cell seeded microbeads may be optionally subjected to a further cell culturing step in suspension, so as to increase the extracellular matrix deposition within the microbeads.
  • step (b) of the process is carried out following the seeding of the microbeads or after the optional step of culturing in suspension the microtissues obtained by seeding the cells onto the microbeads.
  • step (b) consists of injecting the microtissues in a porous microtube that allows nutrient flow to the cells and directionality (geometrical guidance) for the neotissue growth.
  • the porosity of the microtube allows the exchange of nutrients and waste products of the cellular metabolism between the culture medium and the cells and vice versa. Furthermore, the geometry of the microtube guides cell growth, inducing the cells to grow along the longitudinal axis of the microtube and leading to formation of the desired fiber structure. Indeed, it has been demonstrated that directional cell locomotion plays a crucial role in many physiological processes underlying tissue development and organization (Chun-Nin et Al. Biophys. J. Vol 79:144-152, 2000).
  • the microtube may have an inner diameter from 300 ⁇ to 500 ⁇ .
  • the microtube may be made with a polymeric material, for example polytetrafluoroethylene (PTFE).
  • PTFE polytetrafluoroethylene
  • PTFE is suitable since it is nonreactive, chemically and thermally stable, so that it may be also subjected to sterilization by autoclaving.
  • microtissues are injected in the microtube, said microtube is cultured under cell culture conditions in order to allow the cells in the microtissues to continue their growth, extracellular matrix d formation of the bioengineered fiber.
  • the culture conditions employed in step (c) may be dynamic or static conditions, depending on whether the microtube is subjected to stirred or not.
  • the cell growth and extracellular matrix deposition lead to the biological union of the microtissues within the microtube, thus leading to the formation of a continuous structure of biological material shaped as a fiber.
  • the amount of extracellular matrix deposited by the cells depends upon the cell type, the cell culture conditions (presence of stimulating factors), nature of the substrate and number of cells. Therefore, in an embodiment of the present invention, the rate of synthesis of the extracellular matrix may be further enhanced by addition of specific biochemical factors, such as ascorbic acid, preferably in an amount of 50 mg/ ml can be used.
  • specific biochemical factors such as ascorbic acid
  • Formation of the bioengineered tissue may be monitored during time by means of histological analysis, for example by staining with hematoxylin/ eosin (figures 5a and 5b).
  • bioengineered fibers of tissue of animal origin obtainable by means of the process described herein may be used in the field of textile industries.
  • the emulsion is cooled below 20°C and 30 ml of ethanol are added to remove toluene and thus obtain the porous gelatin microbeads.
  • the microbeads are then further washed with ethanol and after a final wash acetone they are dried at room temperature (figure Id).
  • the microbeads are sieved and the fraction between 75 and 150 ⁇ is recovered.
  • Such fraction of microbeads is stabilized by means of chemical treatment with glyceraldehyde, so as to render the gelatin insoluble in water at 37°C.
  • the stabilized microbeads have been examined by meas of Scanning Electron Microscopy and the presence of a large number of cavities having about 20 ⁇ diameter was observed (figure 2).
  • the degradation rate of the gelatin microbeads may be modulated by variation of the degree of crosslinking, i.e. the amount of crosslinks introduced during the stabilization step.
  • the degree of crosslinking i.e. the amount of crosslinks introduced during the stabilization step.
  • two different amounts of glyceraldehyde 10% and 20% w/w of the microbeads
  • the microbeads crosslinked at 20% have been sterilized by autoclaving and the microbeads crosslinked at 10% were sterilized by irradiation with gamma-rays.
  • the microbeads were then placed in contact with the culture medium containing serum before the cell seeding step.
  • Primary fibroblasts were amplified in Petri dishes having a 100 mm diameter and were seeded on both types of microbeads at a concentration of 5xl0 5 cells/mg (figure le). Cell seeding was conducted under dynamic conditions by subjecting the suspension of cells and microbeads to stirring, so as to achieve a uniform distribution of the cells on the microbeads.
  • the dynamic seeding has been carried out in a spinner flask (100 ml Bellco) by using an intermittent stirring for 5 hours in which 5 minutes of stirring were alternated with 30 minutes of static culture, then using continuous stirring for 90 hours.
  • the stirring rate employed was 30 rpm.
  • MTT is a yellow tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide) which is converted into a blue formazan by the dehydrogenases present in living cells.
  • figure 3 depicts a graph of the decreasing concentrations of cells in the culture medium over time.
  • the concentration of cells (p) for each time point is normalized to the initial concentration of cells (p 0 ).
  • the MTT assay revealed a low number of live cells in the microbeads after 2 hours from seeding (figure 4a) which grows over time after 5 hours (figure 4b), 24 hours (figure 4c), 48 hours (figure 4d) and 96 hours (figure 4e), as evidenced by the increasingly deeper blue color of the microbeads. It is thus evident that the gelatine microbeads represent a good substrate for fibroblast adhesion and growth.
  • histological analysis with hematoxylin/eosin staining have been performed on samples taken from cultures after 4 days of seeding.
  • the samples were fixed in formalin (24 hours), sequentially dehydrated in alcohol (75°, 85°, 95°, 100°) and embedded in paraffin. Sections having a 5 ⁇ thickness were obtained with a microtome (Zeiss) and stained with hematoxylin/eosin.
  • the fibroblasts-seeded microbeads were transferred into the porous PTFE microtubes (Markel Corporation ).
  • the microtubes have an inner diameter of 430 ⁇ , an outer diameter of 860 ⁇ and porous surface wherein the pore size is about 6 ⁇ (figures 7a and 7b).
  • the seeded microbeads were transferred to the microtubes by means of an apparatus as shown in figure 8.
  • This apparatus comprises a chamber (1) containing a suspension of the fibroblast-seeded microbeads (2); the chamber is connected to a pump (3) by means of a valve (4) and has an exit (5) connected to a microtube (6) having micropores (7).
  • a stirring device (8) such a magnetic stirrer, may be placed in correspondence of the chamber (1).
  • the pump (3) transfers the suspension of seeded microbeads (2) from the chamber (1) to the microtube (6) through exit (5).
  • the end of the PTFE microtube is closed to maintain the microbeads inside the microtube while allowing the medium to flow trough the porosity of the microtube.
  • the microtubes filled with the seeded microbeads were then kept under static culture conditions in the presence of ascorbic acid to stimulate the extracellular matrix deposition. After 7 days of culturing it was possible to remove from the PTFE microtube a bioengineered fiber having a length of 6 cm and a diameter of 400 ⁇ .
  • the fiber consists of a structure of extracellular matrix, fibroblasts and microbeads (figure 9).
  • the assembly of the seeded gelatin microbeads (microtissues) and the extracellular matrix deposition processes led to the establishment of cellular interactions between the fibroblasts and to the assembly of the desired bioengineered fiber.
  • the process according to the invention allows to obtain structures in the form of fibers of tissue of animal origin without the need for subsequent assembly, since the microtubes allow to obtain a biological tissue formed by the cells having the desired shape.

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Abstract

La présente invention a pour objet un procédé de préparation d'une fibre de tissu d'origine animale modifiée biologiquement comprenant les étapes consistant : a) à ensemencer des cellules eucaryotes sur des microperles poreuses biodégradables présentant une vitesse de dégradation régulée et accordable dans des conditions d'ensemencement dynamiques, ce qui permet d'obtenir des microtissus ; b) à injecter les microtissus dans un microtube poreux approprié à garantir une alimentation nutritive aux cellules et l'orientation du tissu synthétisé ; et (c) à cultiver le microtube rempli des microtissus, ce qui permet d'obtenir une fibre modifiée biologiquement. En outre, la présente invention concerne une fibre de tissu d'origine animale modifiée biologiquement pouvant être obtenue au moyen dudit procédé. La présente invention concerne aussi l'utilisation de ladite fibre modifiée biologiquement en tant que fil dans l'industrie textile.
PCT/IT2009/000488 2009-10-28 2009-10-28 Fibre de tissu animal modifiée biologiquement in vitro et son utilisation dans l'industrie textile Ceased WO2011051983A1 (fr)

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US12600862B2 (en) 2020-05-01 2026-04-14 Modern Meadow, Inc. Protein polyurethane alloys and layered materials including the same

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WO2008061261A2 (fr) * 2006-11-17 2008-05-22 Cytograft Tissue Engineering, Inc. Préparation et utilisation de filetages de synthèse cellulaire

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