Multicomponent biomaterials

Abstract

The present invention describes a biomaterial comprising (a) a collagen composition comprising (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen, and (b) a polymer. The biomaterial is useful in the manufacture of engineered biomaterials, which may be leather-like materials.

Description

[Technical field]

[0001] The present invention relates to a multi-component biomaterial comprising a collagen composition and a polymer, and to a method for producing such a biomaterial. [Background technology]

[0002] Leather is a widely used material and there is a huge global demand for leather products, for example, leather is used for furniture upholstery, clothing, shoes, luggage, handbags and accessories.

[0003] Natural leather is produced by tanning animal hides and skins, often cowhide. Animal hides (and therefore leather made from animal hides) are primarily made up of collagen, a fibrous protein. Collagen is a general term for a family of at least 28 different collagen types, all of which consist of the -(Gly-XY) n -, so that about one third of the amino acid residues in collagen are glycine. X is often proline and Y is often hydroxyproline. Thus, collagen's structure can consist of triple units wrapped around peptide chains of different lengths. The triple helices can be linked together in bundles called fibrils, or fibril bundles can join together to create fibers. Collagen fibers typically join together throughout the layers of the skin. Crosslinks or links can provide strength to the material.

[0004] The properties of natural leather are influenced by the type of animal hide used. In particular, different animals may produce collagen with different amino acid compositions resulting in different properties. Variations in collagen structure also occur throughout the thickness of the hide. The outer upper surface of the hide is generally composed of a fine mesh of collagen fibrils, while the deeper portion (also known as the dermis) is composed of larger fiber bundles. The outer upper surface of the leather is smoother and softer than the dermis. Thus, to produce natural leather with a smooth outer side on both sides, it is necessary to combine two outer pieces of the dermis side together and sew or glue them together. To avoid this combining step, a leather material is needed in which the collagen structure can be controlled to produce a smooth surface on both sides.

[0005] The post-processing steps used in leather production are also limited by the natural variations in collagen structure between different animal hides. Although the final properties of leather can be controlled to some extent by incorporating stabilizing and lubricating molecules into the hide during the tanning stage, the selection of these molecules is limited by the need to penetrate the dense structure of the hide. A method is needed to produce leather materials in which the original collagen structure of the hide does not limit the post-processing steps that can be used.

[0006] Alternative methods of making leather-like materials known in the art include culturing collagen to produce sheets that can then be crosslinked to produce leather-like materials. However, such methods are typically not very efficient and difficult to implement in large-scale manufacturing. Furthermore, leather-like materials made purely from collagen are typically not very strong. Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, there is a need to develop new biomaterials that can be processed to produce improved leather-like biomaterials, and methods for creating leather-like biomaterials. [Means for solving the problem]

[0008] The inventors have found that a collagen composition comprising (i) at least 30% by weight partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen, can be used in combination with a polymer or a combination of polymers to create an improved biomaterial. This biomaterial can provide an improved leather-like material compared to those known in the art. Previously known biomaterials made from collagen compositions use only collagen in the collagen composition and generally require time-consuming and resource-intensive processing steps, for example to create large volumes of acidic solvent. The manufacture of such biomaterials made only from collagen also typically requires the handling of very thick and viscous collagen gels. This has the disadvantage that bubbles may form in the gel during processing that are difficult to remove, which can lead to defects in the material. Previously known biomaterials also typically have low tensile strength. These factors mean that the biomaterials may be unsuitable for further processing into leather-like biomaterials with high strength and smooth appearance. However, the inventors have found that biomaterials made from collagen compositions containing (i) at least 30% by weight partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen are stronger than previously known biomaterials made from collagen compositions containing collagen alone. Furthermore, the biomaterials can be manufactured more efficiently, using fewer resources, and bubbles can be more easily removed during the manufacturing process. Furthermore, the use of a polymer or combination of polymers means that the mechanical properties (e.g., stiffness, strength, flexibility) of the biomaterial can be more precisely controlled than biomaterials that do not contain a polymer. In particular, the use of a polymer or combination of polymers typically results in biomaterials with improved flexibility compared to biomaterials that do not contain a polymer.

[0009] Accordingly, the present invention provides a biomaterial comprising a dehydrated collagen gel, the collagen gel comprising: (a) a collagen composition and (b) a polymer; The collagen composition comprises: (i) partially hydrolyzed collagen; and (ii) collagen and / or fully hydrolyzed collagen; A biomaterial is provided, wherein the collagen composition comprises at least 30% by weight partially hydrolyzed collagen.

[0010] The present invention also provides a method for producing a biomaterial, comprising the steps of: a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial A method is provided that includes:

[0011] Also provided is a leather-like engineered biomaterial comprising the biomaterial described herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Collagen Composition The biomaterial of the present invention comprises a collagen composition and a dehydrated collagen gel formed from a polymer or a combination of polymers. As used herein, a collagen composition is any composition that comprises collagen or any collagen derivative (e.g., partially hydrolyzed collagen or fully hydrolyzed collagen). In the present invention, the collagen composition comprises (i) partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen.

[0013] As used herein, collagen refers to collagen in triple helical structure. Collagen may be acid soluble collagen. Partially hydrolyzed collagen refers to a single strand of triple helical collagen. Partially hydrolyzed collagen may include gelatin. Typically, as used herein, partially hydrolyzed collagen is gelatin. Fully hydrolyzed collagen refers to collagen peptides and / or amino acids. Fully hydrolyzed collagen may include collagen hydrolysates. Typically, as used herein, fully hydrolyzed collagen is collagen hydrolysates. Partially hydrolyzed collagen and / or fully hydrolyzed collagen may be acid soluble.

[0014] Collagen, partially hydrolyzed collagen and fully hydrolyzed collagen can each independently be derived from any animal source or product. Alternatively, collagen, partially hydrolyzed collagen and / or fully hydrolyzed collagen can be prepared by in vitro synthesis techniques. Alternatively, collagen, partially hydrolyzed collagen and / or fully hydrolyzed collagen can be obtained from fungi (e.g. yeast) or bacteria, for example, using fermentation techniques. As used herein, collagen or collagen derivatives derived from a particular animal, fungal or bacterial source or product (e.g. partially hydrolyzed collagen or fully hydrolyzed collagen) refer to collagen-containing components extracted as part of a collagen composition from an animal, fungal or bacterial source or product, which are optionally further processed (e.g., hydrolyzed or purified) to produce collagen or collagen derivatives. For example, partially hydrolyzed collagen derived from an animal, fungal or bacterial source or product is originally extracted as part of a collagen composition from an animal source or product, and then obtained by hydrolysis of that collagen composition.

[0015] For example, collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen may each independently be derived from aquatic, bovine, or porcine products, preferably aquatic or porcine products. As used herein, collagen (or partially hydrolyzed or fully hydrolyzed collagen) derived from aquatic, bovine, or porcine sources or products may also be referred to as aquatic, bovine, or porcine collagen (or partially hydrolyzed or fully hydrolyzed collagen), respectively. Typically, at least one of the partially hydrolyzed collagen, collagen, and fully hydrolyzed collagen composition is derived from aquatic products. For example, in one embodiment, the partially hydrolyzed collagen is partially hydrolyzed marine collagen. In one embodiment, the fully hydrolyzed collagen is fully hydrolyzed marine collagen. In one embodiment, the collagen is marine collagen. In one embodiment, the collagen is not bovine collagen. In one embodiment, neither the collagen, the partially hydrolyzed collagen, nor the fully hydrolyzed collagen is derived from a bovine product. The collagen, partially hydrolyzed collagen, and / or fully hydrolyzed collagen in a particular collagen composition may all be derived from the same type of animal source, or from different types of animal sources. For example, in one embodiment, the collagen composition includes partially hydrolyzed marine collagen and porcine collagen.

[0016] Animal source or product can be any part of animal that contains collagen.For example, aquatic product can be any part of aquatic animal that contains collagen.As used herein, aquatic animal can be any animal that exists mainly or exclusively in an aqueous environment, including animals found in freshwater environment and ocean.Aquatic animal can be fish, such as bass, bream, brill, European catshark, catfish, black cod, cod, small flounder, dogfish, eel, flounder, darter, haddock, halibut, mackerel, plaice, pollock, ray, salmon, sardine, skate, star shark, plaice, tilapia, or tuna.Aquatic animal can be invertebrate, such as sea anemone, clam, coral, hydroid, jellyfish, mussel, oyster, scallop, sea cucumber, sea slug, conch, sea urchin, sponge, starfish, or worm. The aquatic animal may be an arthropod, such as an arachnid, crustacean, insect, or myriapod.

[0017] The seafood product may be a freshwater seafood product, a saltwater seafood product, an invertebrate product or an arthropod product. In one embodiment, the seafood product is a seafood product, including freshwater and saltwater fish. The seafood product may be any part of a fish that contains collagen. Typically, the seafood product includes one or more of fish skin, fish scales, fish swim bladder, or fish joints and / or tendons, for example, one or more of fish skin, fish scales and / or fish swim bladder. All of these seafood products contain collagen, but the collagen content is particularly high in the fish swim bladder used in one preferred embodiment. The seafood product includes fish skin in another preferred embodiment. Seafood products are more sustainable than similar collagen-containing products from other animals, such as bovine products. This is because less water is required for the production of seafood products and less carbon emissions are produced. In particular, seafood skin is a conveniently available waste product, and therefore the use of seafood skin has environmental benefits.

[0018] The bovine product may be any part of a bovine animal that contains collagen. In one embodiment, the bovine product is bovine tendon. The bovine animal may be a cow, bison, buffalo or antelope. Typically, the bovine product is a cow product. The porcine product may be any part of a porcine animal that contains collagen. In one embodiment, the porcine product is pig skin. The porcine animal may be a pig, a hog or a boar.

[0019] In the present invention, the collagen composition comprises at least 30% by weight of partially hydrolyzed collagen. The presence of partially hydrolyzed collagen (e.g., gelatin) in the collagen composition improves the strength of the biomaterial and any processed biomaterials made from the biomaterial. As used herein, reference to weight percent of collagen or collagen derivative means the weight of collagen or collagen derivative expressed as a weight percentage of all collagen or collagen derivative components in the collagen composition.

[0020] The collagen composition may comprise at least 40% by weight, at least 50% by weight, at least 60% by weight, at least 70% by weight, at least 75% by weight, at least 80% by weight, at least 85% by weight, at least 90% by weight, or at least 95% by weight of partially hydrolyzed collagen. Preferably, the collagen composition comprises at least 70% by weight, at least 75% by weight, at least 80% by weight, at least 85% by weight, or at least 90% by weight of partially hydrolyzed collagen, more preferably at least 70% by weight or at least 80% by weight of partially hydrolyzed collagen. Typically, the collagen composition comprises less than 99% by weight of partially hydrolyzed collagen, for example 95% by weight or less of partially hydrolyzed collagen. The collagen composition may comprise 30-95% by weight, 50-95% by weight, 60-95% by weight, 70-95% by weight, 75-95% by weight, or 80-95% by weight of partially hydrolyzed collagen. Alternatively, the collagen composition may comprise 30-90%, 50-90%, 60-90%, 70-90%, 75-90% or 80-90% by weight of partially hydrolyzed collagen, or 30-85%, 50-85%, 60-85%, 70-85%, 75-85% or 80-85% by weight of partially hydrolyzed collagen.

[0021] The collagen composition comprises collagen and / or fully hydrolyzed collagen. The collagen composition may comprise at least 1% by weight of collagen and / or one or a mixture of fully hydrolyzed collagen. Typically, the collagen composition comprises at least 2%, at least 3%, at least 4% or at least 5% by weight of collagen and / or one or a mixture of fully hydrolyzed collagen, preferably at least 5% by weight. The collagen composition may comprise at least 10%, at least 15%, at least 20%, at least 30%, at least 40% or at least 50% by weight of collagen and / or one or a mixture of fully hydrolyzed collagen. The collagen composition may comprise, in total, 70% by weight or less, 60% by weight or less, 50% by weight or less, 40% by weight or less, 30% by weight or less, 25% by weight or less, 20% by weight or less, 15% by weight or less or 10% by weight or less of collagen and / or fully hydrolyzed collagen. Typically, the collagen composition comprises 5-70%, 5-50%, 5-40%, 5-30%, 5-25% or 5-20% by weight of collagen and / or fully hydrolyzed collagen or a mixture thereof. Alternatively, the collagen composition may comprise 10-70%, 10-50%, 10-40%, 10-30%, 10-25% or 10-20% by weight of collagen and / or fully hydrolyzed collagen or a mixture thereof. Alternatively, the collagen composition may comprise 15-70%, 15-50%, 15-40%, 15-30%, 15-25% or 15-20% by weight of collagen and / or fully hydrolyzed collagen or a mixture thereof.

[0022] The collagen composition may comprise at least 1% by weight collagen and / or at least 1% by weight fully hydrolyzed collagen. The collagen composition may comprise at least 2%, at least 3%, at least 4% or at least 5% by weight collagen and / or fully hydrolyzed collagen. In one embodiment, the collagen composition comprises at least 5% by weight collagen and / or at least 5% by weight fully hydrolyzed collagen.

[0023] When the collagen composition contains collagen, the collagen composition may contain at least 1 wt%, at least 5 wt%, at least 10 wt%, at least 15 wt%, at least 20 wt%, or at least 25 wt%, preferably at least 5 wt% or at least 10 wt% collagen. Typically, when the collagen composition contains collagen, the collagen composition contains 70 wt% or less collagen, for example, 60 wt% or less, 50 wt% or less, 40 wt% or less, 30 wt% or less, 25 wt% or less, 20 wt% or less, 15 wt% or less, or 10 wt% or less collagen. Preferably, the collagen composition contains 50 wt% or less collagen. The collagen composition may contain 30 wt% or less collagen. Typically, the collagen composition contains 5 to 70 wt%, 5 to 50 wt%, or 5 to 40 wt%, or 5 to 30 wt%, or 5 to 25 wt%, or 5 to 20 wt% collagen. Preferably, the collagen composition contains 5 to 50 wt%, or 5 to 30 wt% collagen. Alternatively, the collagen composition may comprise 10-70%, 10-50%, 10-40%, 10-30%, 10-25%, or 10-20% collagen by weight. Alternatively, the collagen composition may comprise 15-70%, 15-50%, 15-40%, 15-30%, 15-25%, or 15-20% collagen by weight. In one embodiment, the collagen composition does not contain collagen. It has been found by the inventors that limiting the amount of collagen in the collagen composition allows the composition to contain more partially hydrolyzed collagen, thereby improving the tensile strength of the biomaterial and increasing the efficiency of the manufacturing process (particularly by reducing or eliminating the required neutralization and improving the handling of the product during manufacture, particularly by reducing the viscosity of the product and thereby reducing bubble formation). The use of less collagen also reduces the cost of the resulting biomaterial.

[0024] When the collagen composition contains fully hydrolyzed collagen, the collagen composition may comprise at least 1 wt%, at least 5 wt%, at least 10 wt%, at least 15 wt%, at least 20 wt%, or at least 25 wt% fully hydrolyzed collagen. The presence of fully hydrolyzed collagen (e.g., collagen hydrolysate) in the collagen composition may improve the hardness, elasticity, ductility, and strength of the biomaterial and any processed biomaterial made from the biomaterial. However, it is important to control the amount of fully hydrolyzed collagen in the collagen composition. This is because a large amount of fully hydrolyzed collagen weakens the collagen gel and the resulting biomaterial, and the collagen gel does not bind well. When the collagen composition contains fully hydrolyzed collagen, typically the collagen composition comprises no more than 50 wt%, no more than 40 wt%, no more than 30 wt%, no more than 25 wt%, no more than 20 wt%, no more than 15 wt%, or no more than 10 wt% fully hydrolyzed collagen. Preferably, the collagen composition comprises no more than 30 wt% or no more than 20 wt% fully hydrolyzed collagen. Typically, the collagen composition comprises 5-50% by weight, or 5-40% by weight, or 5-30% by weight, or 5-25% by weight, or 5-20% by weight of fully hydrolyzed collagen. Preferably, the collagen composition comprises 5-30% by weight or 5-20% by weight of fully hydrolyzed collagen. Alternatively, the collagen composition may comprise 10-50% by weight, 10-40% by weight, 10-30% by weight, 10-25% by weight, or 10-20% by weight of fully hydrolyzed collagen. Alternatively, the collagen composition may comprise 15-50% by weight, 15-40% by weight, 15-30% by weight, 15-25% by weight, or 15-20% by weight of fully hydrolyzed collagen. In one embodiment, the collagen composition does not contain fully hydrolyzed collagen.

[0025] In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% by weight partially hydrolyzed collagen, and (ii) 5-70% by weight collagen and / or fully hydrolyzed collagen. In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% by weight partially hydrolyzed collagen, and (ii) 5-50% by weight collagen and / or fully hydrolyzed collagen. In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% by weight partially hydrolyzed collagen, and (ii) 5-40% by weight collagen and / or fully hydrolyzed collagen. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-95% by weight partially hydrolyzed collagen, and (ii) 5-30% by weight collagen and / or fully hydrolyzed collagen. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen, and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-90% by weight partially hydrolyzed collagen, and (ii) 10-30% by weight collagen and / or fully hydrolyzed collagen.

[0026] polymer In the present invention, the collagen gel comprises a polymer in addition to the collagen composition.

[0027] The polymer may comprise a biopolymer or a biobased polymer. In one embodiment, the polymer comprises a biopolymer. In one embodiment, the polymer is a biopolymer. In one embodiment, the polymer comprises a biobased polymer. In one embodiment, the polymer is a biobased polymer.

[0028] As used herein, a biopolymer is any polymer produced by the cells of an organism. For example, a biopolymer can be a polymer produced by a plant, a microorganism, or an animal. A biopolymer is usually biodegradable. Typically, a biopolymer is a polypeptide, a protein, a polynucleotide, a polysaccharide, a polymer of isoprene, a gum, a polyphenol polymer, a lipid, a polymer of fatty acids, or a pigment. In one embodiment, the biopolymer is a polysaccharide or a gum. In one embodiment, the biopolymer is a polysaccharide.

[0029] As used herein, a bio-based polymer is a synthetic polymer that includes materials where at least a portion of the polymer is produced from a non-petrochemical source (from a renewable feedstock). Typically, a bio-based polymer is composed of materials produced from a non-petrochemical source (from a renewable feedstock). Bio-based polymers are typically non-biodegradable.

[0030] The polymer may be water soluble. Alternatively, the polymer may be water insoluble.

[0031] Polymers include cellulose, nanocellulose, lignin, starch, alginates, gum arabic, guar gum, gellan gum (including low acyl gellan gum and high acyl gellan gum), xanthan gum, carrageenan (including kappa carrageenan, iota carrageenan, and lambda carrageenan), polyhydroxyalkanoates (PHAs), polyhydroxy fatty acids (PHFs), bacterial cellulose, hyaluronan, hyaluronic acid, curdlan, pullulan, silk (including silk fibroin and silk fibers), elastin, chitin, chitosan, casein, whey, alginates, cellulose, cellulose acetate, cellulose esters ... The polymer may comprise one or a mixture of polymers selected from: bumin, polyisoprene (rubber), keratin, poultry feather fiber (PFF), mucin, pectin, agar, agarose, actin, fibrin, fibrinogen, suberin, cutin, melanin, cotton, collagen fibers, polylactic acid (PLA), wool, polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polytrimethylene terephthalate, polyurethane, polycarbonate, poly(ether-ester) copolymers, polyamides, polyesteramides, unsaturated polyesters, epoxy resins and phenolic resins. In one embodiment, the polymer comprises one or a mixture of polymers selected from cellulose, nanocellulose, lignin, starch, alginate, gum arabic, guar gum, gellan gum (low acyl and high acyl), xanthan gum, carrageenan (including kappa carrageenan, iota carrageenan, lambda carrageenan), polyhydroxyalkanoate (PHA), PHF, bacterial cellulose, hyaluronan, hyaluronic acid, curdlan, pullulan, silk (including silk fibroin and silk fiber), elastin, chitin, chitosan, casein, whey, albumin, polyisoprene (rubber), keratin, PFF, mucin, pectin, agar, agarose, actin, fibrin, fibrinogen, suberin, cutin, melanin, cotton, collagen fiber, and wool. As used herein, cellulose may be microcellulose or nanocellulose. Preferably, the cellulose is microcellulose.

[0032] Typically, the polymer comprises starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, guar gum, xanthan gum, cellulose, nanocellulose, collagen fibers, keratin, cotton, or mixtures thereof. In one embodiment, the polymer comprises starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, cellulose, nanocellulose, collagen fibers, keratin, cotton, or mixtures thereof. In one embodiment, the polymer comprises starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, guar gum, xanthan gum, cellulose, nanocellulose, keratin, cotton, or mixtures thereof. In one embodiment, the polymer comprises starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, cellulose, nanocellulose, keratin, cotton, or mixtures thereof. The polymer may comprise starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, guar gum, xanthan gum, or mixtures thereof. The polymer may comprise starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, or mixtures thereof. Alternatively, the polymer may comprise cellulose, nanocellulose, collagen fibers, keratin, cotton, or mixtures thereof. In one embodiment, the polymer comprises cellulose, nanocellulose, keratin, cotton, or mixtures thereof. The polymer may comprise cellulose. In one embodiment, the polymer does not comprise collagen.

[0033] The water insoluble polymer is typically present in the collagen gel in the form of a reinforcing material, for example as fibers (including short or continuous fibers), thin films, flakes or plates, segments, particles, fillers, whiskers or spheres. For example, the polymer may be present in the collagen gel in the form of fibers, typically fibers of a biopolymer. In one embodiment, the polymer comprises cellulose fibers, nanocellulose fibers, cotton fibers, silk fibers, collagen fibers, polylactic acid (PLA) fibers, wool fibers, keratin fibers, or mixtures thereof. In one embodiment, the polymer comprises cellulose fibers, nanocellulose fibers, cotton fibers, silk fibers, collagen fibers, wool fibers, keratin fibers, or mixtures thereof. In one embodiment, the polymer comprises cellulose fibers, nanocellulose fibers, cotton fibers, silk fibers, wool fibers, keratin fibers, or mixtures thereof. In one embodiment, the polymer comprises cellulose fibers, collagen fibers, keratin fibers, cotton fibers, or mixtures thereof. The polymer may comprise collagen fibers. In one embodiment, the polymer comprises nanocellulose fibers, collagen fibers, keratin fibers, cotton fibers, or mixtures thereof. In one embodiment, the polymer is selected from nanocellulose fibers, collagen fibers, keratin fibers, and cotton fibers. In one embodiment, the polymer comprises cellulose fibers, nanocellulose fibers, keratin fibers, cotton fibers, or mixtures thereof. As used herein, cellulose fibers may be microcellulose fibers or nanocellulose fibers. Preferably, the cellulose fibers are microcellulose fibers.

[0034] When the polymer comprises collagen, the collagen is present in the form of a reinforcing material, for example collagen fibers. In one embodiment, the polymer comprises collagen reinforcing material (for example collagen fibers) in an amount of 15% or less. The polymer may comprise collagen reinforcing material (for example collagen fibers) in an amount of 10% or less by weight or 5% or less by weight, for example 2% or less by weight or 1% or less by weight, compared to the total weight of the collagen composition. In another embodiment, the polymer does not comprise collagen.

[0035] Typically, a collagen gel contains one polymer. However, a gel may contain more than one polymer (i.e., a combination of polymers), for example, two, three, or four different polymers. A gel may contain two polymers. For example, a collagen gel may contain starch and pectin, or cellulose fibers and wool fibers. When a collagen gel contains more than one polymer, each polymer may be present in equal amounts or in different amounts.

[0036] The presence of a polymer in a collagen gel is useful in the manufacture of biomaterials. In particular, the polymer or combination of polymers provides increased control over the mechanical properties of the biomaterial and any engineered biomaterials produced from it, compared to biomaterials that do not contain a polymer. For example, the polymer may allow for control of the stiffness of the material.

[0037] Combination of Polymer and Collagen Compositions The biomaterials of the present invention include dehydrated collagen gels, wherein the gel comprises (a) a collagen composition and (b) a polymer. The polymer (or combination of polymers) and collagen composition may be bonded by chemical bonds (e.g., chemical crosslinking), or by physical means (e.g., by being physically entangled or intertwined), or by a combination of the two.

[0038] In one embodiment, the collagen composition and the polymer(s) are linked by chemical crosslinks. Alternatively, the collagen composition and the polymer(s) may be physically entangled.

[0039] The combination of the water-soluble polymer and the collagen composition may be referred to herein as a polymer blend. In the polymer blend, the polymer and collagen composition are homogeneous, i.e., form a single phase. In one embodiment, the collagen gel comprises a polymer blend comprising the collagen composition and a polymer. In one embodiment, the collagen gel comprises a polymer blend comprising the collagen composition and a polymer, where the collagen composition and the polymer are chemically crosslinked.

[0040] The combination of the water-insoluble polymer and the collagen composition may be referred to herein as a composite. In the composite, the polymer and the collagen composition are heterogeneous, i.e., separate phases. In such composites, the polymer is typically present as a reinforcing material, for example, as a fiber (including short or continuous fibers), a thin film, a flake or plate, a segment, a particle, a filler, a whisker, or a sphere. In one embodiment, the collagen gel comprises a composite comprising the collagen composition and a polymer. In one embodiment, the collagen gel comprises a composite comprising the collagen composition and a polymer, where the collagen composition and the polymer are physically entangled and / or chemically crosslinked.

[0041] The nature of the interaction between the collagen composition and the polymer(s) depends on, for example, the type of polymer used, as well as the type of crosslinker, crosslinking conditions, and the solubility of the polymer. For example, a chemically crosslinked material can be formed, in which the polymer has functional groups that can react with the crosslinker to form chemical crosslinks between the polymer and the collagen composition. If the polymer does not contain any functional groups suitable for crosslinking, the polymer and the collagen composition are typically physically entangled. Those skilled in the art will easily understand how to achieve chemical crosslinking and / or physical entanglement for a particular polymer.

[0042] When two or more types of polymers are used in the collagen gel, the polymers can be combined with the collagen composition in the same way or in different ways.For example, the collagen gel can include a first polymer that is chemically crosslinked with the collagen composition and a second polymer that is physically entangled with the collagen composition and the first polymer.Alternatively, both the first polymer and the second polymer can be chemically crosslinked or physically entangled with the collagen composition.

[0043] Collagen gel typically comprises at least 1% by weight of polymer compared to the weight of collagen composition. Collagen gel may comprise at least 2% by weight, at least 5% by weight, at least 10% by weight, at least 15% by weight, at least 20% by weight, at least 25% by weight, at least 30% by weight, at least 40% by weight, or at least 50% by weight of polymer compared to the weight of collagen composition. Preferably, collagen gel comprises at least 2% by weight, or at least 5% by weight of polymer compared to the weight of collagen composition.

[0044] The collagen gel may comprise 50% or less, 40% or less, 30% or less, 25% or less, 20% or less, 15% or less, 10% or less, 5% or less, 2% or less, or 1% or less by weight of polymer relative to the weight of the collagen composition. Preferably, the collagen gel comprises 25% or less or 20% or less by weight of polymer relative to the weight of the collagen composition.

[0045] The collagen gel may contain 1-50% by weight, or 1-40% by weight, or 1-30% by weight, or 1-25% by weight, or 1-20% by weight, or 1-15% by weight, or 1-10% by weight, or 1-5% by weight of the polymer, relative to the weight of the collagen composition. Alternatively, the collagen gel may contain 2-50% by weight, or 2-40% by weight, or 2-30% by weight, or 2-25% by weight, or 2-20% by weight, or 2-15% by weight, or 2-10% by weight, or 2-5% by weight of the polymer, relative to the weight of the collagen composition. Alternatively, the collagen gel may contain 5-50% by weight, or 5-40% by weight, or 5-30% by weight, or 5-25% by weight, or 5-20% by weight, or 5-15% by weight, or 5-10% by weight of the polymer, relative to the weight of the collagen composition. Preferably, the collagen gel comprises 1% to 25% by weight, or 1 to 20% by weight, or 2 to 20% by weight, or 2 to 15% by weight, or 2 to 10% by weight of polymer compared to the weight of the collagen composition.

[0046] When the collagen gel comprises two or more polymers, the collagen gel may comprise each polymer within the weight ranges outlined above, or the collagen gel may comprise a total amount of polymers within the weight ranges outlined above.

[0047] In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% by weight partially hydrolyzed collagen and (ii) 5-70% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-25% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% by weight partially hydrolyzed collagen and (ii) 5-70% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% by weight partially hydrolyzed collagen and (ii) 5-70% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% partially hydrolyzed collagen and (ii) 5-70% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% polymer by weight relative to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 30-95% partially hydrolyzed collagen and (ii) 5-70% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% polymer by weight relative to the weight of the collagen composition.

[0048] In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% by weight partially hydrolyzed collagen and (ii) 5-50% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-25% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% by weight partially hydrolyzed collagen and (ii) 5-50% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% by weight partially hydrolyzed collagen and (ii) 5-50% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% partially hydrolyzed collagen and (ii) 5-50% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% polymer by weight relative to the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 50-95% partially hydrolyzed collagen and (ii) 5-50% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% polymer by weight relative to the collagen composition.

[0049] In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% by weight partially hydrolyzed collagen and (ii) 5-40% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-25% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% by weight partially hydrolyzed collagen and (ii) 5-40% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% by weight partially hydrolyzed collagen and (ii) 5-40% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-20% by weight polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% partially hydrolyzed collagen and (ii) 5-40% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% polymer by weight relative to the collagen composition. In one embodiment, the collagen composition comprises one or a mixture of (i) 60-95% partially hydrolyzed collagen and (ii) 5-40% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% polymer by weight relative to the collagen composition.

[0050] In one preferred embodiment, the collagen composition comprises (i) 70-95% by weight of partially hydrolyzed collagen, and (ii) 5-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 1-25% by weight of polymer, relative to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70-95% by weight of partially hydrolyzed collagen, and (ii) 5-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 1-20% by weight of polymer, relative to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70-95% by weight of partially hydrolyzed collagen, and (ii) 5-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 2-20% by weight of polymer, relative to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-95% partially hydrolyzed collagen and (ii) 5-30% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% polymer by weight relative to the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-95% partially hydrolyzed collagen and (ii) 5-30% collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% polymer by weight relative to the collagen composition.

[0051] In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-25% by weight polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1-20% by weight polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-20% by weight polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% by weight polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 80-95% by weight partially hydrolyzed collagen and (ii) 5-20% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% by weight polymer compared to the weight of the collagen composition.

[0052] In one preferred embodiment, the collagen composition comprises (i) 70-90% by weight of partially hydrolyzed collagen, and (ii) 10-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 1-25% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70-90% by weight of partially hydrolyzed collagen, and (ii) 10-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 1-20% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70-90% by weight of partially hydrolyzed collagen, and (ii) 10-30% by weight of collagen and / or fully hydrolyzed collagen, or a mixture thereof, and the collagen gel comprises 2-20% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-90% by weight partially hydrolyzed collagen and (ii) 10-30% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-15% by weight polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises one or a mixture of (i) 70-90% by weight partially hydrolyzed collagen and (ii) 10-30% by weight collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2-10% by weight polymer compared to the weight of the collagen composition.

[0053] Extracted collagen composition In one embodiment, the collagen gel may be formed from a collagen composition extracted from an animal product, particularly an aquatic product. As used herein, a collagen composition extracted from an animal (e.g., an aquatic animal) product may be described as an extracted collagen composition. As used herein, a collagen composition extracted from an aquatic product may be described as a marine collagen composition. The aquatic products referred to herein are as described above.

[0054] Advantageously, collagen compositions extracted from marine products can be efficiently used to create biomaterials that are well suited for further processing steps to create leather-like biomaterials. Previously known methods of producing biomaterials from collagen do not use marine products as a collagen source and often require long and complicated extraction steps to provide collagen in a form suitable for further processing. Cultured collagen has also been used, but this is not efficient and the process is not easily scalable. Cultured collagen also has the disadvantage of not providing an endogenous mixture of natural collagen proteins.

[0055] The collagen in the extracted collagen composition can be extracted using acid (i.e., acid-soluble collagen) or pepsin (i.e., pepsin-soluble collagen). The collagen in the extracted collagen composition can also be extracted using an alkaline solution. Partially and / or fully hydrolyzed collagen can be added to the extracted collagen composition to obtain an extracted collagen composition having a greater amount of partially and / or fully hydrolyzed collagen. For example, partially hydrolyzed collagen and / or fully hydrolyzed collagen can be added to the extracted collagen composition to provide a collagen composition containing (i) at least 30% by weight of partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen. The preferred amounts of collagen and / or fully hydrolyzed collagen are as described above. In one embodiment, the collagen composition as defined herein comprises an extracted collagen composition, for example, a collagen composition extracted from a marine product.

[0056] The presence of partially and / or fully hydrolyzed collagen in the extracted collagen composition is useful for the production of biomaterials. In particular, partially and / or fully hydrolyzed collagen can increase the efficiency of cross-linking and gel formation. Furthermore, partially and / or fully hydrolyzed collagen can improve the properties of biomaterials and any processed biomaterials produced from the biomaterials. In particular, the presence of partially hydrolyzed collagen can result in softer and more elastic biomaterials and / or processed biomaterials compared to biomaterials that do not contain partially hydrolyzed collagen.

[0057] The extracted collagen composition typically comprises collagen, and optionally partially and / or completely hydrolyzed collagen. Typically, the extracted collagen composition comprises at least one of acid-soluble collagen, partially hydrolyzed collagen, and completely hydrolyzed collagen. As used herein, acid-soluble collagen is collagen that can be extracted using acid. The extracted collagen composition can contain at least 20% by weight, at least 30% by weight, or at least 40% by weight of acid-soluble collagen. For example, the extracted collagen composition can contain 20-50% by weight of acid-soluble collagen. The extracted collagen composition can contain at least 1% by weight, at least 5% by weight, at least 10% by weight, at least 15% by weight, at least 20% by weight, or at least 30% by weight of partially hydrolyzed collagen. For example, the extracted collagen composition can contain 1-40% by weight of partially hydrolyzed collagen. The extracted collagen composition can contain at least 1% by weight, at least 5% by weight, at least 10% by weight, at least 15% by weight, at least 20% by weight, or at least 30% by weight of completely hydrolyzed collagen. For example, the extracted collagen composition can contain 1-40% by weight of fully hydrolyzed collagen. In one embodiment, the extracted collagen composition contains at least 20% by weight of acid-soluble collagen, and / or at least 1% by weight of partially hydrolyzed collagen, and / or at least 1% by weight of fully hydrolyzed collagen. In one embodiment, the extracted collagen composition contains at least 1% by weight of partially hydrolyzed collagen, and / or at least 1% by weight of fully hydrolyzed collagen. The amounts provided above for acid-soluble collagen can also be applied to collagen.

[0058] The extracted collagen composition may be an endogenous composition, i.e., contains collagen, collagen derivatives (e.g., partially and / or completely hydrolyzed collagen), and other components (e.g., naturally occurring impurities) as found naturally in the marine product. For example, collagen typically has the telopeptide region intact. This may make products formed from the extracted collagen composition more desirable to certain consumer groups. Advantageously, the collagen composition extracted from the marine product may be used in its endogenous form without the need for complex processing. Furthermore, the endogenous composition may contain collagen derivatives, which may improve the efficiency of biomaterial production, and advantageously affect the properties of any processed biomaterials produced from the biomaterial.

[0059] Manufacturing method The present invention also relates to a method of producing the biomaterial described herein. Typically, the method of producing the biomaterial comprises: a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial Includes.

[0060] The collagen composition can be any collagen composition as defined herein. In one embodiment, the present invention provides a method for producing a biomaterial, comprising the steps of: a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial the collagen composition comprising (i) partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen; The method is provided wherein the collagen composition comprises at least 30% by weight partially hydrolyzed collagen.

[0061] The polymer may be any polymer or combination of polymers as defined herein.

[0062] In one embodiment, the forming step (a) comprises contacting the collagen composition and the polymer(s) with one or more crosslinking agents to form a crosslinkable collagen mixture, and crosslinking the crosslinkable collagen mixture to form a collagen gel. Typically, the polymer is added to the crosslinkable collagen mixture before crosslinking occurs. However, the polymer may also be added to a partially crosslinked crosslinkable collagen mixture, i.e., after crosslinking has already occurred to some extent.

[0063] In some embodiments, the crosslinks form between the collagen composition and the polymer. Alternatively or additionally, the crosslinks form within the collagen composition and around the polymer such that the collagen composition and polymer are physically entangled.

[0064] The forming step can include adding a fatliquoring component and / or a dye or pigment to the collagen composition, crosslinkable collagen mixture, or polymer. In one embodiment, the present invention provides a method for producing a biomaterial of the present invention, comprising the steps of: a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial wherein the forming step comprises adding a fatliquoring component and / or a dye or pigment to the collagen composition.

[0065] In one embodiment, the present invention provides a method for producing a biomaterial of the present invention, comprising the steps of: a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial wherein the forming step includes contacting the collagen composition and the polymer with one or more crosslinking agents to form a crosslinkable collagen mixture, and crosslinking the crosslinkable collagen mixture to form a collagen gel, and the forming step further includes adding a fatliquoring component and / or a dye or pigment to the collagen composition, the crosslinkable collagen mixture, or the polymer.

[0066] Also provided herein is a method for producing a biomaterial, comprising the steps of: a) extracting a collagen composition from an animal product, preferably a marine product; and b) forming a collagen gel comprising the collagen composition and a polymer, and dehydrating the collagen gel to form a biomaterial. wherein the collagen composition comprises collagen, optionally partially and / or fully hydrolyzed collagen.

[0067] The extraction step (a) may also include adding partially hydrolyzed and / or fully hydrolyzed collagen to the extracted collagen composition to provide a collagen composition comprising (i) partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen, the collagen composition comprising at least 30% by weight of partially hydrolyzed collagen. In one embodiment, the collagen composition comprises partially hydrolyzed collagen, collagen, and optionally fully hydrolyzed collagen, the collagen composition comprising at least 30% by weight of partially hydrolyzed collagen.

[0068] Any of the methods described herein can further include a step of treating the biomaterial to form a processed biomaterial. The processed biomaterial can be a leather-like biomaterial. The treatment step(s) can include one or more of drying, dyeing, fatliquoring, finishing, and coating the biomaterial.

[0069] Any of the methods described herein may further include adding a textile backing to the collagen gel, biomaterial, or processed biomaterial. Suitable textiles are known to those skilled in the art and include cotton, wool, silk, leather, suede, flax, jute, hemp, felt, linen, fiberglass cloth, nylon, polyester, Tencel, rayon, Kevlar, viscose, spandex, acrylic, polyamide, and mixtures thereof. Adding the textile backing may include attaching a textile backing layer to the biomaterial or processed biomaterial. For example, the textile backing may be added by laminating an adhesive layer to the biomaterial / processed biomaterial and / or textile backing layer via a coating method. Suitable coating methods are known to those skilled in the art and include, for example, spray coating, brush coating, knife-over-roller coating, roll-to-roll coating, reverse roller coating, curtain coating, and slot die coating. The lamination process may use lamination rollers (as part of a roll-to-roll process) or lamination lines (lamination is done using pressure and / or heat to bond the backing layer to the collagen gel / biomaterial / processed biomaterial, usually via an adhesive; this adhesive may itself be on a roll). Alternatively, the textile backing layer may be placed on or in the crosslinkable collagen mixture prior to crosslinking (or placed on or in the partially crosslinked crosslinkable collagen mixture), for example by partially or completely immersing the textile in the crosslinkable collagen mixture, and then bonded to the biomaterial by crosslinking during the (remaining) crosslinking step. The textile backing may also be added by casting the biomaterial or processed biomaterial onto the textile backing layer.

[0070] In one embodiment, the extraction step (a) comprises washing the animal product, e.g., aquatic product, with an alkaline solution, optionally further washing the aquatic product with a degreasing agent, contacting the aquatic product with an acidic solution having a pH of 4-5, and obtaining an extracted collagen composition from the acidic solution; the forming step (b) comprises contacting the extracted collagen composition and a polymer with one or more crosslinking agents to form a crosslinkable collagen mixture, crosslinking the crosslinkable collagen mixture to form a collagen gel, and dehydrating the collagen gel; optionally, the method further comprises a processing step (c) to form a processed biomaterial after the forming step (b) to form a processed biomaterial, the processing step comprises fatliquoring the biomaterial, staining the fatliquored biomaterial, drying the stained and fatliquored biomaterial, and mechanically processing the dried biomaterial.

[0071] Also described herein is a biomaterial obtainable by the methods described herein. The biomaterial can be a leather-like biomaterial.

[0072] (a) Extraction The following section describes the extraction of collagen compositions from animal products. References herein to animal products can be interpreted as references to the particular animal product (e.g., seafood) from which collagen is extracted.

[0073] Collagen can be extracted from animal products, e.g., seafood products, by contacting the animal products with an acidic solution. The acidic solution can be any solution with a pH below 7. Typically, the acidic solution can have a pH between 3 and 6, preferably between 4 and 6. In a preferred embodiment, the acidic solution has a pH between 4 and 5. The acidic solution can include any weak acid or diluted strong acid with an appropriate pH. Typically, the acidic solution is an aqueous solution of acetic acid, formic acid, or hydrochloric acid. In one embodiment, the acidic solution is an aqueous solution of acetic acid.

[0074] The animal product may be contacted with the acidic solution for at least 6 hours, or at least 12 hours, or at least 24 hours. Typically, the animal product is contacted with the acidic solution for about 12 hours. The animal product may be contacted with the acidic solution at a temperature of less than 30° C., or less than 20° C., or less than 10° C. Typically, the animal product is contacted with the acidic solution at a temperature of about 4° C. After contacting the animal product with the acidic solution, the extracted collagen composition may be separated from the solution. The extracted collagen composition may be freeze-dried (lyophilized).

[0075] The extraction process may further include washing the animal product with an alkaline solution before contacting the animal product with the acidic solution. The animal product may be washed more than once with the alkaline solution. For example, the animal product may be washed twice with the alkaline solution. The alkaline solution may be useful for removing non-collagenous proteins from the animal product and for breaking down the animal product. The alkaline solution may be any solution with a pH greater than 7. Typically, the alkaline solution is an aqueous solution of sodium hydroxide, potassium hydroxide, sodium carbonate, or magnesium carbonate. In one embodiment, the alkaline solution is a solution of sodium hydroxide.

[0076] The extraction process may further include washing the animal product with a degreasing agent prior to contacting the animal product with the acidic solution. The degreasing agent may be useful for removing fat from the product. Typically, the degreasing agent is an alcohol solution, an organic solvent (e.g., chloroform, petroleum ether, or n-hexane) or supercritical CO2. In a preferred embodiment, the degreasing agent is an alcohol solution. The alcohol solution may contain less than 70% v / v, less than 50% v / v, or less than 30% v / v alcohol in water. Typically, the alcohol solution contains between 5-20% v / v alcohol in water. Using an alcohol solution in water rather than pure alcohol prevents dehydration of the animal product and reduces the efficiency of collagen composition extraction. The alcohol may be methanol, ethanol, propan-1-ol, propan-2-ol (isopropyl alcohol), butan-1-ol, or butan-2-ol. In one embodiment, the alcohol solution is a solution of isopropyl alcohol.

[0077] When the animal product is washed with both an alkaline solution and a degreaser prior to contacting the animal product with an acidic solution, the washing with the alkaline solution can occur before or after washing with the degreaser. Typically, the animal product is washed with an alkaline solution before washing with the degreaser. In one embodiment, the animal product is washed with an alkaline solution before washing with the alcohol solution.

[0078] The animal product can be washed with water before and / or after each portion of the extraction process, for example, before contacting the animal product with the alkaline solution, between contacting the animal product with the alkaline solution and the degreasing agent, and between contacting the animal product with the degreasing agent and the acidic solution.

[0079] The extraction methods described herein do not require additional processing steps, such as enzymatic digestion or purification of the collagen composition. This reduces the time and resources required to perform the extraction compared to methods that require such steps. Also, the collagen protein is retained in an undigested form, without fragmentation of the protein chains.

[0080] (b) formation The collagen composition can be or include an extracted collagen composition, which is formed into a collagen gel with a polymer or combination of polymers, and the collagen gel is dehydrated to form the biomaterial.

[0081] The collagen composition is typically first provided in a suitable solution for forming a biomaterial. For example, the collagen composition can be diluted in water or a buffer, or the lyophilized collagen composition can be dissolved in water or a buffer. A suitable concentration is 1-200 mg / mL, for example, 10-100 mg / mL of collagen protein (including collagen, acid-soluble collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen).

[0082] Advantageously, the collagen composition used in the present invention may include an extracted collagen composition, which is typically highly soluble in aqueous solutions at a pH of 5-8, such as pH 6-8, pH 6-7 or about pH 7. This means that a small volume of aqueous solvent (e.g. water) can be used without the need to add a large volume of acid to lower the pH and dissolve the collagen composition. In one embodiment, the collagen composition has a solubility of at least 20 mg / mL, at least 30 mg, or at least 40 mg, or at least 50 mg of collagen composition per mL of aqueous solvent at a pH of 5-8 and a temperature of 25° C. Preferably, the collagen composition has a solubility of at least 20 mg / mL, at least 30 mg, or at least 40 mg, or at least 50 mg of collagen composition per mL of aqueous solvent at a pH of 6-7 and a temperature of 25° C. For collagen compositions containing collagen, the addition of an acidic solvent may be required to achieve dissolution. Any suitable weak acid or diluted strong acid, such as an aqueous solution of acetic acid, formic acid, or hydrochloric acid, may be used. Typically, collagen is dissolved in an acidic solvent separately into partially hydrolyzed and / or fully hydrolyzed collagen, and then the collagen solution is added to the partially hydrolyzed and / or fully hydrolyzed collagen solution to obtain a mixed solution of collagen composition with a pH of 5-8, preferably 6-8, or 6-7. Collagen can be dissolved in an aqueous solution with a pH of less than 5, less than 4, less than 3, or less than 2, preferably less than 3. In one embodiment, collagen is dissolved in an aqueous solution with a pH of about 2. However, collagen compositions containing small amounts of collagen require only small amounts of acidic solution. Furthermore, the overall collagen composition is typically still soluble in solutions with a pH between 5 and 8. Physical agitation, such as stirring, mixing, and sonication, can also be used to aid dissolution. High temperatures (i.e., temperatures above about 25°C) can also be used to aid dissolution. The use of a highly soluble collagen composition in this process is beneficial because a smaller volume of solvent is required and it is easier to remove the solvent during dehydration.

[0083] This is in contrast to collagen mixtures that are commonly used in known processes for producing biomaterials and contain only collagen or mostly collagen. Such collagen mixtures are typically insoluble at pH between pH 5 and 8 and require a very low pH to dissolve, e.g., a pH below pH 5. The use of collagen compositions that do not require highly acidic conditions for dissolution means that the solution is easily neutralized after the gel formation process. This improves production efficiency and further reduces the total amount of solvent required.

[0084] The polymer or combination of polymers is typically added to the solution of the collagen composition. If the polymer is water-soluble, it can be dissolved in the solution together with the collagen composition using the same techniques as described above. Alternatively, the polymer can be dissolved separately from the collagen composition, and then the solution of the polymer and the solution of the collagen composition are combined. If the polymer is water-insoluble, it is typically added to the solution of the collagen composition after the collagen composition is completely dissolved.

[0085] As part of the formation process, the collagen composition, which may be or may include an extracted collagen composition, and optionally the polymer (if the polymer is capable of forming chemical crosslinks), can be crosslinked using any suitable protein crosslinking method known in the art. Typically, the collagen composition and the polymer(s) are contacted with one or more crosslinking agents to form a crosslinkable collagen mixture. The crosslinking agent(s) can be any molecule with bifunctional, trifunctional or polyfunctional reactive groups capable of forming crosslinks between collagen molecules. Alternatively, the crosslinking agent(s) can be a molecule that can be used in a photoinitiated crosslinking process. The crosslinking agent can be an enzyme. Typically, the crosslinker(s) are one or more agents selected from alcohols, aldehydes, amines, azides, carboxylic acids, carbodiimides, chromium salts, epoxides, hydrazides, isocyanates, sulfhydryls, N-hydroxysuccinimide esters, imiodesters, maleimides, haloacetyls, pyridyl disulfides, aryl azides, diazirines, aglycones, and Staudinger ligation pairs. Typically, the one or more crosslinkers may include glutaraldehyde, transglutaminase, carbodiimides, hydrazides, or genipin. For example, the one or more crosslinkers may include glutaraldehyde and / or transglutaminase. Suitable amounts of crosslinkers are known to those skilled in the art. Typically, 0.1-40 w / w%, for example 1-10 w / w%, of crosslinker may be used based on the total weight of the collagen composition. When glutaraldehyde is used as a crosslinking agent, the amount of glutaraldehyde in the crosslinkable composition can be, for example, 0.5-10 w / w%, for example, 1-5 w / w%. Alternatively, when the crosslinking agent is an enzyme, typically, the enzyme is used in an amount of 0.1-70 U per gram of collagen composition, for example, 0.1-40 U per gram of collagen composition, 1-60 U / g, 10-60 U / g, 20-60 U / g, or 30-60 U / g. The amount of crosslinking agent can be 1-10 U / g. When transglutaminase is used as a crosslinking agent, the amount of transglutaminase can be, for example, 0.5-10 U / g, for example, 1-5 U / g.Alternatively, the amount of transglutaminase may be 5-60 U / g, for example 10-60 U / g or 20-60 U / g.

[0086] The cross-linkable collagen mixture may also contain a dye or pigment. Thus, in one embodiment, a dye or pigment is added to the collagen composition, the cross-linkable collagen mixture, or the polymer. For example, the dye or pigment may be an aqueous dye or pigment, an alcoholic dye or pigment, an acid dye, a direct dye, a mordant dye, or a base dye. In one embodiment, the dye is an aqueous dye or an aqueous pigment. The use of a dye in this step may help ensure that the resulting biomaterial is uniformly stained throughout its thickness.

[0087] The crosslinkable collagen mixture can also contain a fatliquoring component, such as a fatliquoring emulsion. Thus, in one embodiment, a fatliquoring component is added to the collagen composition, the crosslinkable collagen mixture, or the polymer. The fatliquoring emulsion can include fatty salts, such as sulfonates, sulfites, and / or phosphates of tri-glycerides. The use of a fatliquoring emulsion in this step can improve the depth, speed, and uniformity of fatliquoring penetration through the biomaterial compared to fatliquoring after gel formation, while still providing beneficial softening and water repellency properties. Fatliquoring is further described in the description of the processing steps.

[0088] The crosslinkable collagen mixture may contain one or more further additives, such as one or more plasticizers. The plasticizers help to soften and make the resulting biomaterial flexible. Suitable plasticizers are known to those skilled in the art. In one embodiment, the plasticizer is glycerol and the crosslinkable collagen mixture comprises glycerol. The plasticizer may be used, for example, in an amount of 5-50 w / w% based on the weight of the collagen composition. When using glycerol (also known as glycerin) as a plasticizer, the amount of glycerol in the crosslinkable composition may be, for example, 10-40 w / w%, such as 20-40 w / w%.

[0089] One or more antifoaming agents can also be added to the crosslinkable collagen mixture such that the collagen composition (and crosslinkable collagen mixture) further comprises one or more antifoaming agents. Antifoaming agents are also called defoaming agents. Antifoaming agents remove bubbles or foam formed in the composition, improving the handling of the composition and aiding in the creation of a more biomaterial. Typically, physical agitation, such as stirring or agitation, is used in combination with the antifoaming agent to ensure complete elimination of bubbles and foam. In one embodiment, physical agitation can be applied to the collagen composition and / or crosslinkable collagen mixture containing an antifoaming agent for at least 10 minutes, at least 20 minutes, at least 30 minutes, or at least 1 hour. Crosslinkable collagen mixtures containing partially hydrolyzed collagen are easier to defoam using an antifoaming agent than mixtures containing collagen only. For example, the inventors have attempted to reproduce Example 2 of EP3205668, which describes the formation of bioprocessed leather from bovine collagen. It was found that the crosslinkable collagen mixture was very thick with a large amount of bubbles that could not be easily removed by conventional methods (e.g., sonication) even at pH 2. It is believed that the presence of partially hydrolyzed collagen in the collagen used in the present invention produces a thinner and more viscous mix, and that an antifoaming agent is used to aid in defoaming.

[0090] Suitable antifoaming agents are known to those skilled in the art. In one embodiment, the antifoaming agent is a food grade antifoaming agent. The antifoaming agent can be a silicone-based emulsion, a polypropylene glycol composition, or an ethylene oxide (EO) and propylene oxide (PO) copolymer. In one embodiment, the antifoaming agent is a silicone-based emulsion. The antifoaming agent can be used in an amount of, for example, 0.001 w / w% to 5 w / w%, where w / w% means the weight of the active component of the antifoaming agent per weight of the total solution. Typically, the amount of antifoaming agent added to the solution depends on the amount of foam generated in the formation process, which can be affected by the different molecular weights of collagen extracted from different sources. The antifoaming agent is preferably added in an amount sufficient to remove at least 70%, at least 80%, at least 90% or at least 95% of the bubbles and foam. Typically, the defoaming agent is used in an amount of 0.001 w / w% to 5 w / w%, or 0.01 w / w% to 3 w / w%, or 0.1 w / w% to 3 w / w%, or 0.1 w / w% to 2 w / w%, or 0.1 w / w% to 1 w / w%, or 0.1 w / w% to 0.5 w / w%. Preferably, the defoaming agent is used in an amount of 1 w / w% or less. For example, the defoaming agent can be used in an amount of 0.001 w / w% to 1 w / w%, or 0.01 w / w% to 1 w / w%, or 0.1 w / w% to 1 w / w%. When a silicone-based emulsion is used as the defoaming agent, the amount of the silicone-based emulsion in the crosslinkable composition can be, for example, 0.001 w / w% to 5 w / w%, for example, 0.1 w / w% to 2 w / w%.

[0091] Antifoaming agents can also improve the properties of biomaterials and any processed biomaterials produced from the biomaterials. In particular, low concentrations (e.g., 0.1% to 3% w / w) of antifoaming agents have been found to increase the tensile strength of the resulting biomaterial.

[0092] Typically, the crosslinkable collagen mixture has a pH of 6-8, preferably about 7, prior to crosslinking. If the crosslinkable collagen mixture has a pH outside this range, an appropriate amount of acid or base can be added to achieve the desired pH. As explained above, the collagen composition used in the present invention is advantageously highly soluble in aqueous solutions with a pH of 5-8, preferably a pH of 6-8, or a pH of 6-7. Thus, in one embodiment, no neutralization is required in the forming step to obtain a crosslinkable mixture with a pH of 6-8. In one embodiment, the forming step involves a neutralization step to raise the pH of the crosslinkable mixture to a range of pH 6-8, and the neutralization step involves raising the pH of the crosslinkable mixture by 3 or less, 2 or less, or 1 or less.

[0093] The crosslinkable collagen mixture (including the collagen composition and the polymer) can be crosslinked to form a collagen gel including the collagen composition and the polymer. Crosslinking can be achieved by allowing the crosslinkable collagen mixture to stand for at least 1 second, or at least 10 seconds, or at least 30 seconds, or at least 1 minute, or at least 2 minutes, or at least 5 minutes, or at least 15 minutes. Typically, the crosslinkable collagen mixture is allowed to stand for at least 30 seconds, or at least 1 minute, or at least 2 minutes, or at least 5 minutes. Alternatively, the crosslinkable collagen mixture is allowed to stand for at least 30 minutes, or at least 1 hour, or at least 2 hours. Alternatively, the crosslinkable collagen mixture is allowed to stand for at least 12 hours, or at least 24 hours, or at least 36 hours, or at least 48 hours. In one embodiment, the crosslinkable collagen mixture is allowed to stand for at least 48 hours. The crosslinkable collagen mixture can be allowed to stand at a temperature of about 1°C to about 60°C, for example, about 1°C to about 50°C, about 1°C to about 30°C, about 1°C to about 20°C, or about 1°C to about 10°C. Alternatively, the crosslinkable collagen mixture may be allowed to stand at a temperature of about 15° C. to about 60° C., such as about 20° C. to about 60° C., or about 20° C. to about 50° C., or about 25° C. to about 50° C. In one embodiment, the crosslinkable collagen mixture is allowed to stand at a temperature of about 4° C. In one embodiment, the crosslinkable collagen mixture is allowed to stand at a temperature of about 20° C.

[0094] In some embodiments, the crosslinking step results in the formation of chemical crosslinks between the collagen composition and the polymer. In some embodiments, the crosslinking step results in the formation of crosslinks within the collagen composition, such that a physical network of the crosslinked collagen composition is intertwined or entangled with the polymer. In some embodiments, the crosslinking step results in the formation of crosslinks between the collagen composition and the polymer, as well as the formation of a physically entangled network of the crosslinked collagen composition and the polymer.

[0095] The collagen gel is dehydrated to form a biomaterial. The collagen gel can be dehydrated using a suitable dehydrating solvent that is miscible with water, such as a ketone or an alcohol. Typically, the collagen gel is dehydrated in acetone or ethanol. Dehydration in a dehydrating solvent, such as acetone or ethanol, ensures uniform dehydration throughout the collagen gel, and in particular prevents one side of the collagen gel from drying faster than the other. Alternatively, the collagen gel can be dehydrated using a dehydrator at a temperature of about 25°C to about 100°C, such as about 25°C to about 45°C. Typically, the collagen gel is dehydrated using a dehydrator at a temperature of about 30°C to about 40°C, such as about 35°C. Alternatively, the collagen gel can be dehydrated using a dehydrator at a temperature of about 50°C to about 100°C, such as about 75°C to about 100°C. The collagen gel can be dehydrated in the dehydrator for at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, or at least 12 hours. Typically, the collagen gel is dehydrated in a dehydrator for at least 8 hours, for example about 10 hours. Dehydration in a dehydrator reduces the amount of solvent used in the manufacturing process compared to dehydration processes using solvents. The water content of the dehydrated collagen gel is typically between 1-35%, for example between 10-35%.

[0096] The shape of the dehydrated collagen gel (i.e., biomaterial) is not limited and may include any two-dimensional or three-dimensional shape. The shape of the biomaterial can be controlled, for example, by crosslinking the crosslinkable collagen mixture in a suitably shaped mold. Alternatively, the collagen gel can be shaped and / or reshaped before and / or after dehydration using suitable shaping techniques. Such shaping can involve bending, folding, stretching, rolling or cutting the collagen gel or the dehydrated collagen gel. The biomaterial can be formed by continuous manufacturing methods such as coating. Suitable coating methods include, but are not limited to, knife-over-roller coating (KOR coating), slot-die coating, roller-on-roller coating, curtain coating, dip coating, flow coating, spray coating and brushing. Coating methods include coating the collagen gel onto a carrier or substrate (such as a paper, polymer or fabric carrier or substrate) directly or with an adhesive layer, followed by removal of the carrier or substrate after dehydration. Alternatively, the collagen gel may be coated onto a polymer or biopolymer layer already coated onto the carrier or substrate.

[0097] Typically, the biomaterial is formed into a sheet, and thus the biomaterial is provided in the form of a sheet. The sheet can be any thickness, but typically the sheet is less than 5 cm thick. Typically, the sheet can be less than 3 cm, 2 cm, 1 cm, 0.5 cm, or 0.1 cm thick. The sheet can be of uniform thickness, or the sheet can have different thicknesses. The sheet can be formed by placing the crosslinkable collagen mixture in a suitable mold and crosslinking to provide a desired thickness and allow gel formation. Alternatively, the sheet can be formed by a continuous manufacturing method, such as coating. The sheet can be composed of a single layer of biomaterial, or it can be composed of multiple layers of biomaterial, for example, multiple layers of biomaterial with different properties. The sheet can be composed of multiple layers of the same biomaterial, or two or more layers of biomaterial can have different compositions.

[0098] The collagen gel formed after crosslinking can be temporarily frozen to allow for easy removal from the mold, and is then typically thawed prior to dehydration.

[0099] The biomaterial according to the present invention typically has a high tensile strength. For example, the biomaterial can have a tensile strength of at least 5 MPa, or at least 10 MPa, at least 15 MPa, at least 20 MPa, or at least 25 MPa. Preferably, the biomaterial has a tensile strength of at least 10 MPa, at least 15 MPa, or at least 20 MPa. In one embodiment, the biomaterial has a tensile strength of about 5 MPa to about 25 MPa. Tensile strength is typically measured according to standard method ISO3376 (2020).

[0100] Furthermore, the biomaterial according to the present invention is typically semi-soft and pliable. The biomaterial generally has a uniform collagen structure throughout its thickness. In addition, the properties of the biomaterial can be easily altered by the amount and type of polymer, crosslinker(s) and any other additives used.

[0101] (c) Processing The biomaterial can be further processed to form a processed biomaterial. Thus, the methods described herein can further comprise a processing step (c) after the sheet-forming step (b) to form a processed biomaterial. The processed biomaterial is typically a leather-like material.

[0102] The processing step may include any step or combination of steps that produces a leather-like material. As used herein, leather-like material refers to a material that has similar physical properties to natural leather. Typically, leather-like materials are strong and flexible. Leather-like materials may not show cracks when the material is folded in two. Typically, the processing step is performed on a biomaterial formed from a dehydrated collagen gel, but at least the processing steps of fatliquoring and / or dyeing can be incorporated into the gel formation process described above. If fatliquoring and / or dyeing steps are included in the gel formation process, the biomaterial formed after dehydration of the collagen gel may be a leather-like material that does not require further processing.

[0103] The leather-like material may have a durability of greater than 3 mm, or greater than 5 mm, or greater than 7 mm. In one embodiment, the leather-like material has a durability of about 5 mm to about 10 mm. Durability refers to the amount of outer expansion and strength of the leather. Durability is typically measured using a lastometer according to standard methods ISO 3379 (2015) or DIN 53325.

[0104] Leather-like materials may have light fastness such that no change in color or surface deterioration is observed after 10 hours, 20 hours, 30 hours, or 40 hours. Light fastness is typically measured according to standard method ISO 105-B02 (2014). Light fastness may also be measured using light of wavelengths between 300 and 400 nm.

[0105] The leather-like material may be resistant to environmental aging such that no color change or surface deterioration is observed after 20 hours, 40 hours, or 60 hours of being subjected to an accelerated environmental aging test. In one embodiment, the accelerated environmental aging test may include subjecting the biomaterial to a temperature of 60±2° C. and a humidity of 90±5% RH.

[0106] The leather-like material may have color fastness to water spot such that no change in shade or surface deterioration is observed after 10 hours, 13 hours, or 16 hours. Color fastness to water spot is typically measured according to standard method ISO 15700 (1998).

[0107] The leather-like material may have a "Martindale" abrasion resistance of at least 3000 cycles, at least 4000 cycles, or at least 5000 cycles (all measured below 9 kPa). In one embodiment, the leather-like material has a "Martindale" abrasion resistance of about 3000 to about 6000 cycles at less than 9 kPa, as measured using a Martindale abrasion machine. "Martindale" abrasion resistance is typically measured according to standard method ISO 17076-2 (2011).

[0108] The leather-like material may have a "Veslic" color fastness to rubbing such that no deterioration is observed after 100 cycles of wetting and 100 cycles of drying, or 125 cycles of wetting and 125 cycles of drying, or 150 cycles of wetting and 150 cycles of drying. "Veslic" color fastness to rubbing is typically measured according to standard method ISO 11640 (2018).

[0109] The leather-like material may have a tensile strength of at least 5 MPa, or at least 10 MPa, at least 15 MPa, at least 20 MPa, or at least 25 MPa. In one embodiment, the leather-like material has a tensile strength of about 5 MPa to about 25 MPa. Tensile strength is typically measured according to standard method ISO 3376 (2020). The elongation of the leather-like material is measured in the same manner as the tensile strength.

[0110] The leather-like material may have a tear strength of at least 5 N, or at least 10 N, or at least 15 N. In one embodiment, the leather-like material has a tear strength of about 10 to about 20 N. Tear strength is typically measured according to standard methods ISO 3377-1 (2011) or ISO 3377-2 (2016). In one embodiment, tensile tear strength can be measured using a low inertia automated tensile tester at a traverse speed of 300±10 mm / min.

[0111] The leather-like material can have flex resistance such that no surface degradation is observed after 9000 flex cycles, after 12000 flex cycles, or after 15000 flex cycles. Flex resistance is typically measured using a Bally flexometer according to standard method ISO 5402-1 (2017).

[0112] The leather-like material may have a low heat shrinkage, for example less than 50% heat shrinkage, less than 40% heat shrinkage, or less than 30% heat shrinkage. The amount of heat shrinkage is measured by subjecting the sample to 80°C for 15 minutes. The test specimens before the heat shrinkage test are typically 20mm x 30mm. The area of ​​the sample before and after the test is measured and the shrinkage is calculated as follows: Shrinkage percentage (%) = (1-final area / initial area)*100

[0113] The leather-like material may have low levels of chemical impurities. For example, the leather-like material may contain less than the following amounts of one or more of the following impurities: 75 ppm formaldehyde, 1 ppm chlorophenols, 1 ppm total metals, 3 ppm Cr(VI), 200 ppm dicyclohexyl phthalate (DCHP), 0.1 ppm dimethyl fumarate, 30 ppm azo dyes, 1 ppm polycyclic aromatic hydrocarbons, 1 ppm phthalates, 1 ppm Substances of Very High Concern (SVHC) as defined by the European Chemicals Agency, 1 ppm organotin compounds.

[0114] The processing step may include one or more of the steps of fatliquoring, dyeing, and drying the biomaterial. Typically, processing step (c) includes fatliquoring, dyeing, and drying. In one embodiment, the biomaterial is fatliquored, then the fatliquored biomaterial is dyed, then the dyed and fatliquored biomaterial is dried. In another embodiment, the biomaterial is fatliquored, then the fatliquored biomaterial is dried. The processing step may also include a step of mechanically processing the biomaterial. As used herein, a step of mechanically processing the biomaterial may include a step of bending, folding, and / or rolling the biomaterial. In one embodiment, the biomaterial is first fatliquored, then the fatliquored biomaterial is dyed, then the dyed and fatliquored biomaterial is dried, then the dried biomaterial is processed.

[0115] Fatliquorization is the process of fixing fats, oils and / or waxes to the fibers in a material by coating the material with an emulsion of fats, oils and / or waxes in a solvent. Typically, the fats, oils and / or waxes are oils, such as vegetable oils, castor oil, pine oil, lanolin or fish oil. For example, fatliquorization may involve contacting the biomaterial or processed biomaterial with an emulsion of vegetable oil in acetone. In the treatment of natural leather, fatliquorization is used to re-grease the surface of the leather to increase softness and flexibility. Fatliquorization also adds water repellency properties. Fatliquorization of biomaterials produced from marine collagen compositions may produce a stiff and brittle material. However, this stiff and brittle fatliquorized material can still be made into a leather-like material by further processing steps, in particular by further dyeing, drying, treatment with alcohol solutions and / or mechanically working the material.

[0116] Any suitable dye or pigment can be used for dyeing. Suitable leather dyes and pigments are known in the art and may include aqueous dyes and pigments, alcohol-based dyes and pigments, acid dyes, direct dyes, mordant dyes or base dyes. In one embodiment, the dye is an alcohol-based dye, particularly an ethanol-based dye. Treatment with an alcohol solution can be used in addition to or instead of the dyeing process. Typically, the alcohol solution is an ethanol solution.

[0117] The drying step typically involves drying the biomaterial at a temperature of at least 30° C., or at least 40° C., or at least 50° C. Drying may help to soften the rigid biomaterial produced after fat-liquidization. The drying step may be carried out in a dehydrator. Typically, the moisture content of the dried biomaterial may be between 5 and 25%.

[0118] Typically, the treatment also includes a finishing and / or coating step that imparts the desired aesthetics to the biomaterial. Suitable finishing chemicals and formulations are known to those skilled in the art and may include water repellent chemicals, beeswax, or synthetic polymers. If a water-based finishing formulation is used, the finished biomaterial must be dried as described above to remove the aqueous solvent.

[0119] Treatment may also include any other process typically applied to natural leather, including rehydrating, splitting, shaving, neutralizing, filling, setting, conditioning, softening, or buffing.

[0120] Manufacturing of goods The biomaterial or processed biomaterial can be used to manufacture an article comprising the biomaterial or processed biomaterial. The manufacture can include any process of shaping and / or cutting the biomaterial or processed biomaterial. The article can be any article that can be advantageously made from leather-like materials, such as accessories, shoes and furniture.

[0121] [Example] Examples 1-10 relate to biomaterials made from the collagen composition. Examples 11 and 12 relate to biomaterials comprising the collagen composition and a polymer.

[0122] [Example 1] A leather-like engineered biomaterial was made according to the following method: The biomaterial is made from a collagen composition.

[0123] (a) Extraction Cod skin (80 g, wet mass) was cut into 3 cm x 5 cm strips and cleaned with deionized water. 250 mL of 0.1 M NaOH was added and stirred for 2 hours at room temperature. (NaOH was replaced after 1 hour). Skin was washed with water.

[0124] 10% v / v isopropyl alcohol in water (200 mL) was added and stirred for 1 hour at room temperature. The skins were washed with water.

[0125] 200 mL of 1M AcOH was added and stored overnight at 4° C. The extract was then separated. Another 200 mL of 1M AcOH was added to the fish skin and again stored overnight at 4° C.

[0126] The two extracts were combined and freeze-dried to give a white amorphous solid. Yield: 100 g / kg (dried mass of fish skin).

[0127] (b) formation 100 mg of fish skin extract from step (a) was dissolved in 2 mL of water at room temperature. 30 w / w% glycerol and 2 w / w% glutaraldehyde were added and mixed well. The solution was transferred into a mold and left at 4° C. for 2 days for gel formation.

[0128] The gel was placed in a freezer for 1 hour and removed from the mold. After thawing, the gel was placed in 50 mL of acetone for dehydration (shaking table 40 rpm, 48 hours, replaced with fresh acetone after 24 hours).

[0129] After dehydration, the material was semi-soft and pliable.

[0130] c) Processing The material was subjected to fatliquoring. The sample was immersed in a solution of 20% v / v vegetable oil in acetone and placed on a shaking table at 40 rpm for 8 hours. The sample was removed from the solution and the excess amount of oil solution was wiped off. After fatliquoring, the material became very rigid.

[0131] The fatliquored material was then dyed by immersion in ethanolic black leather dye for 1 hour. After dyeing, the excess amount of dye was wiped off.

[0132] The dyed material was then dried in a spin dryer at 40° C. for 5 hours.

[0133] The sample became leather-like after multiple flexing.

[0134] [Example 2] The biomaterial was produced according to the method of Example 1, except that the extraction step (a) was omitted and 100% fish gelatin was used as the starting material for step (b) instead of the fish skin extract.

[0135] After completion of process step (c), the resulting material was softer and more elastic than the material produced in Example 1.

[0136] [Example 3] A biomaterial was prepared according to the method of Example 1, except that the dyeing process of the process step (c) was omitted. It was observed that the stiff biomaterial prepared after fattification had a reduced stiffness after drying.

[0137] [Example 4] A biomaterial was prepared according to the method of Example 1, except that the staining process of the treatment step (c) was combined with the formation step (b). In this example, an aqueous dye is mixed with the collagen solution before gel formation. The resulting biomaterial was observed to be stained evenly throughout the thickness of the biomaterial.

[0138] [Example 5] The following examples describe methods of making biomaterials using partially hydrolyzed collagen, and optionally fully hydrolyzed collagen, but without the use of collagen.

[0139] Partially hydrolyzed collagen and fully hydrolyzed collagen were purchased from Louis Francois and InnerVita.

[0140] Gel formation The protein mixture (2.5 g) was dissolved in 50 mL of degassed type II water at a temperature of around 50-60°C. After complete dissolution, the protein mixture was sonicated for 30 seconds and cooled to room temperature (20-25°C). 0.75 g of glycerin (Intralabs) was added to the protein mixture and stirred for 1-2 minutes until completely dissolved. 0.2 ml of glutaraldehyde (Alfa Aesar, 2 w / w% protein) was added to the solution at room temperature and the solution was mixed for 1-2 minutes before being poured into a 12 cm x 12 cm or 25 cm x 25 cm mold. The hydrogel formed after 30 minutes at room temperature.

[0141] dehydration The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0142] [Example 6] The following examples describe methods of making biomaterials using partially hydrolyzed collagen, and optionally fully hydrolyzed collagen, along with collagen.

[0143] Lyophilized type I collagen sheets isolated from porcine skin and bovine tendon were purchased from Wuxi BIOT Bio-technology Co. Ltd. Marine collagen was extracted from cod skin according to established procedures. Partially hydrolyzed collagen and fully hydrolyzed collagen were purchased from Louis Francois and InnerVita. XIAMETER AFE-1530 antifoam agent (silicone-based emulsion) was purchased from Dow Chemical Company.

[0144] Gel formation 0.25-0.5 g of collagen sheet was cut into small pieces and weighed for use. Partially hydrolyzed collagen and / or partially hydrolyzed collagen / fully hydrolyzed collagen mixture (2-2.25 g) was dissolved in 25 mL of type II degassed water at 50-60 °C. The protein solution was cooled to room temperature (20-25 °C). The prepared collagen pieces were added to 25 mL of 0.01 M aqueous HCl solution. After collagen was fully dissolved, 0.75 g of glycerin was added to the solution. To defoam the solution, 0.2-0.4% (w / w) of antifoam emulsion was added and the solution was further stirred for at least 30 min until bubbles were completely eliminated. The protein solutions were combined and then neutralized by adding an aliquot of 5 M aqueous NaOH solution. 0.2 ml of glutaraldehyde (2 w / w%) was added dropwise and the solution was stirred for 1-2 min before being poured into 12 cm x 12 cm or 25 cm x 25 cm molds. Hydrogels were formed after 30 min at room temperature.

[0145] dehydration The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0146] [Example 7] Biomaterials using various collagen-containing components were prepared according to the methods of Examples 5 and 6. The method of Example 5 was used when the composition did not contain collagen, and the method of Example 6 was used when the composition contained collagen. The tensile strength of the biomaterials was tested using an Instron Model 34SC-05 tensile strength tester according to standard method ISO 3376:2020.

[0147] The composition and tensile strength of the biomaterials are listed in Table 1.

[0148] [Table 1]

[0149] conclusion The results in Table 1 demonstrate that biomaterials produced from collagen compositions containing partially hydrolyzed collagen (entries 3-16) exhibit improved tensile strength compared to biomaterials produced from collagen compositions containing collagen as the only collagen-containing component (entries 1 and 2).

[0150] The results also show that the addition of antifoaming agents at low concentrations successfully prevents the formation of bubbles during the gel formation process, thereby increasing the mechanical strength of the biomaterial.

[0151] Marine collagen is a particularly advantageous collagen source: for example, a biomaterial made from 100% marine collagen (entry 1; 5.4 MPa) showed improved tensile strength over a biomaterial made from 100% bovine collagen (entry 2; 4.1 MPa).

[0152] [Example 8] Biomaterials using various collagen-containing components were prepared according to the methods of Examples 5 and 6. A fat-liquoring emulsion (40 w / w% protein) was added to the collagen composition during gel formation. The results are shown in Table 2.

[0153] [Table 2]

[0154] [Example 9] Biomaterials using various collagen-containing components were prepared according to the methods of Examples 5 and 6, except that transglutaminase (protein-glutamine γ-glutamyltransferase, EC 2.3.2.13) provided by Stabizym TGL-100 was used as the crosslinker instead of glutaraldehyde.

[0155] [Table 3]

[0156] Consideration The addition of antifoam increased the tensile strength of the biomaterials. For example, the biomaterial made from the composition of entry 2 (0.001 w / w% antifoam; 31.5 MPa) showed improved tensile strength over entry 1 (no antifoam; 25.6 MPa).

[0157] [Example 10] Biomaterials using various collagen-containing components were prepared according to the method of Example 9. An aqueous dye (Metropolitan Leather) was added to the solution during gel formation. The results are shown in Table 4.

[0158] [Table 4]

[0159] [Example 11] A biomaterial comprising a chemically crosslinked polymer blend of a collagen composition and a polymer was prepared according to the following method.

[0160] Gel formation Protein mixtures (5 g, 90% partially hydrolyzed collagen, 10% fully hydrolyzed collagen) and potato starch (Sigma Aldrich) at 2%, 5%, 10%, 15%, and 20% (w / w of protein) were dissolved in 50 mL of type II degassed water at a temperature of approximately 50–60 °C. After complete dissolution, the solution was sonicated for 30 s and cooled to room temperature (20–25 °C). 0.75 g of glycerin (Intralabs) was added to the protein mixture and stirred for 1–2 min until completely dissolved. 0.38 ml of glutaraldehyde (Alfa Aesar, 2% (w / w) of protein) was added to the solution at room temperature and the solution was mixed for 1–2 min before being poured into a 12 cm × 12 cm mold. The hydrogel formed after 30 min at room temperature.

[0161] dehydration The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0162] As shown in Table 5, biomaterials containing pectin, alginate, agarose, chitosan, gellan gum and gum arabic were prepared in a similar manner.

[0163] [Table 5]

[0164] [Example 12] A biomaterial comprising a physically entangled composite of collagen composition and polymer was fabricated according to the following method.

[0165] Gel formation The protein mixture (2.5 g, 90% partially hydrolyzed collagen, 10% fully hydrolyzed collagen) was dissolved in 50 mL of type II degassed water at a temperature of approximately 50–60 °C. After complete dissolution, the solution was sonicated for 30 s and cooled to room temperature (20–25 °C). 0.75 g of glycerin (Intralabs) was added to the protein mixture and stirred for 1–2 min until completely dissolved. 0.03 g of cotton fibers (pretreated to remove wax) were dispersed in the solution. 25 U (10 U / g) of transglutaminase was added to the solution. The hydrogel was formed after 30 min at 50 °C.

[0166] dehydration The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0167] Biomaterials containing nanocellulose fibers, collagen fibers and keratin fibers were prepared in a similar manner, as shown in Table 6.

[0168] [Table 6]

[0169] [Example 13] The crosslinkable collagen mixture was placed on a fixed coated paper carrier in front of the knife with a pre-set gap (0.1 mm to 5 mm, typically 1.2 mm). The knife was then actuated such that the blade moved perpendicular to the orientation of the blade edge, so that the mixture was evenly applied to the coated paper in a motion parallel to the orientation of the coated paper. After the samples were coated, they were transferred to a dehydration oven where they were dehydrated at a pre-set temperature for a pre-set time.

[0170] Once dehydration was complete, the samples were removed from the coated paper.

[0171] [Example 14] Biomaterials were made according to the methods described in Example 9 ("control", no polymer) and Example 12 ("cellulose reinforced", cellulose was used as the polymer). The compositions of the two biomaterials before dehydration are as follows:

[0172] [Table 7]

[0173] After dehydration, the biomaterial contained approximately 8% water and 10% cellulose.

[0174] The mechanical properties of the biomaterials are shown in Table 8.

[0175] [Table 8]

[0176] As shown in Table 8, the cellulose-reinforced biomaterials are more flexible than the equivalent biomaterials without the polymer. Additionally, the cellulose-reinforced biomaterials exhibit lower thermal shrinkage and higher tensile strength than the equivalent biomaterials without the polymer. The cellulose-reinforced polymers are also softer than the biomaterials without the polymer.

Claims

1. A biomaterial comprising a dehydrated collagen gel, the collagen gel comprising (a) a collagen composition and (b) a polymer; the collagen composition comprises (i) partially hydrolyzed collagen, and (ii) collagen and / or fully hydrolyzed collagen; A biomaterial, wherein the collagen composition comprises at least 30% by weight of partially hydrolyzed collagen.

2. 2. The biomaterial of claim 1, wherein the collagen composition comprises at least 5% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen.

3. 2. The biomaterial of claim 1, wherein the collagen composition comprises 50 to 95% by weight of partially hydrolyzed collagen.

4. 4. The biomaterial of claim 3, wherein the collagen composition comprises 70 to 95% by weight of partially hydrolyzed collagen.

5. 5. The biomaterial of claim 4, wherein the collagen composition comprises 80 to 95% by weight of partially hydrolyzed collagen.

6. 5. The biomaterial of claim 4, wherein the collagen composition comprises 5 to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen.

7. 6. The biomaterial of claim 5, wherein the collagen composition comprises 5 to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen.

8. 2. The biomaterial according to claim 1, wherein the collagen composition comprises 5 to 30% by weight of fully hydrolyzed collagen, preferably the collagen composition comprises 5 to 20% by weight of fully hydrolyzed collagen.

9. The biomaterial of claim 1 , wherein the collagen gel comprises a polymer blend comprising a collagen composition and a polymer.

10. 10. The biomaterial of claim 9, wherein the polymer is selected from starch, pectin, alginate, chitosan, agarose, gellan gum and gum arabic.

11. The biomaterial of claim 1 , wherein the collagen gel comprises a composite material comprising a collagen composition and a polymer.

12. The biomaterial of claim 11 , wherein the polymer is selected from cellulose fibers, cotton fibers, collagen fibers, and keratin fibers.

13. The biomaterial of claim 12 , wherein the cellulose fibers are microcellulose fibers or nanocellulose fibers.

14. The biomaterial of claim 1 , wherein the collagen composition further comprises an antifoaming agent.

15. 15. The biomaterial of claim 14, wherein the antifoaming agent is present in an amount of 0.001 to 5 w / w%.

16. 16. The biomaterial of claim 15, wherein the antifoaming agent is present in an amount of 0.1 to 2 w / w%.

17. 10. The biomaterial of claim 1, wherein at least one of the partially hydrolyzed collagen, collagen, and fully hydrolyzed collagen is extracted from aquatic products.

18. a) forming a collagen gel comprising a collagen composition and a polymer; and b) Dehydrating the collagen gel to form a biomaterial A method for producing the biomaterial according to any one of claims 1 to 17, comprising:

19. 20. The method of claim 18, wherein the forming step comprises adding a fat-liquoring component and / or a dye or pigment to the collagen composition and / or polymer.

20. 20. The method of claim 18, further comprising processing the biomaterial to form an engineered biomaterial.

21. A leather-like processed biomaterial comprising the biomaterial according to any one of claims 1 to 17.