Multi-component biomaterial

Abstract

Described is a biological material comprising (a) a collagen composition comprising (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen, (b) a polymer, and (c) an additive selected from the group consisting of anti-hydrolysis agents, blowing agents, thickeners, fillers, and flame retardants. The biomaterial may be used to produce a processed biomaterial, which may be a leather-like material.

Description

Technical Field

[0001] This invention relates to multi-component biomaterials comprising collagen compositions, polymers, and additives, and methods for producing said biomaterials. Background Technology

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

[0003] Natural leather is made by tanning animal hides and skin (usually cowhide). Animal hides (and therefore leather made from them) are primarily composed of collagen, a fibrous protein. Collagen is a general term for a family of at least 28 different types of collagen, all characterized by repeating triplets of amino acids – (Gly-XY)…-, so about one-third of the amino acid residues in collagen are glycine. X is usually proline, and Y is usually hydroxyproline. Thus, the structure of collagen can consist of triplet units of peptide chains of varying lengths interwoven. Triple helices can bind together to form bundles called fibrils, and fibril bundles can aggregate together to form fibers. Collagen fibers are typically interconnected throughout the skin layer. Crosslinking, or linking, provides strength to the material.

[0004] The properties of natural leather are influenced by the type of animal hide used. In particular, different animals produce different collagen amino acid compositions, which can lead to different properties. Variations in collagen structure also occur throughout the thickness of the hide. The upper grain of the hide is typically composed of a fine network of collagen fibers, while the deeper layers (also known as the dermis) consist of larger bundles of fibers. The upper grain of the leather is smoother and softer than the dermis. Therefore, to produce natural leather with a smooth texture on both sides, it is necessary to combine two pieces of grain and dermis together and then sew them together or laminate them with an adhesive. A leather material is needed where the collagen structure can be controlled to create a smooth surface on both sides, thus avoiding this combination step.

[0005] The post-processing steps used in leather manufacturing are also limited by the natural variations in collagen structure between different animal hides. Although the final properties of the leather can be controlled to some extent during the tanning stage by incorporating stabilizing and lubricating molecules into the hide, the need for a dense structure that permeates the hide limits the selection of these molecules. 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 for producing leather-like materials known in the art include culturing collagen to produce sheets, which can then be cross-linked to produce a leather-like material. However, this method is generally not very efficient and is difficult to implement on a large scale. Furthermore, leather-like materials made purely from collagen are generally not very strong.

[0007] Therefore, there is a need to develop new biomaterials that can be processed to produce improved leather-like biomaterials, as well as methods for producing leather-like biomaterials. Summary of the Invention

[0008] The inventors have discovered that improved biomaterials can be made by combining a collagen composition comprising (i) at least 30% by weight of partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen with (a) a polymer or polymer combination and (b) an additive or combination of additives selected from anti-hydrolysis agents, foaming agents, thickeners, fillers, and flame retardants. This biomaterial provides an improved leather-like material compared to those known in the art. Previously known biomaterials made from collagen compositions use only the collagen in the composition and generally require time-consuming and resource-intensive processing steps to produce, for example, large amounts of acidic solvents. The manufacture of such collagen-only biomaterials typically also requires processing very thick and viscous collagen gels. This has the disadvantage that air bubbles that are difficult to remove can form in the gel during processing, leading to defects in the material. Previously known biomaterials also typically have low tensile strength. These factors mean that the biomaterials may not be suitable for further processing into leather-like biomaterials with high strength and a smooth appearance. However, the inventors have discovered that biomaterials made from collagen compositions comprising (i) at least 30% by weight of partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen are more robust than previously known biomaterials made from collagen compositions containing only collagen. Furthermore, biomaterials can be produced more efficiently with fewer resources, and air bubbles can be more easily removed during manufacturing. The use of polymers or polymer combinations means that the mechanical properties (e.g., stiffness, strength, flexibility) of the biomaterial can be controlled more precisely compared to biomaterials without polymers. In particular, the use of polymers or polymer combinations generally results in biomaterials with higher flexibility than biomaterials without polymers. Additionally, the use of additives or combinations of additives selected from anti-hydrolysis agents, foaming agents, thickeners, fillers, and flame retardants allows for further control over the properties of the biomaterial. In particular, the use of such additives or combinations of such additives generally results in biomaterials with higher thermal stability and / or humidity stability. The use of additives or combinations of such additives also generally allows for control over the elongation of the biomaterial after tensile strain; for example, additives or combinations of such additives may provide biomaterials with reduced elongation after tensile strain.

[0009] Therefore, the present invention provides a biomaterial comprising a dehydrated collagen gel, wherein the collagen gel comprises (a) a collagen composition, (b) a polymer and (c) an additive;

[0010] The collagen composition includes (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen.

[0011] The collagen composition comprises at least 30% by weight of partially hydrolyzed collagen, and

[0012] The additives are selected from anti-hydrolysis agents, foaming agents, thickeners, fillers and flame retardants.

[0013] A method for producing the biomaterial of the present invention is also provided, the method comprising:

[0014] a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and

[0015] b) Dehydrate the collagen gel to form biomaterials.

[0016] A leather-like processed biomaterial is also provided, which includes the biomaterials described herein. Detailed Implementation

[0017] As used herein, unless otherwise stated, the term “about” means a value of ±10% of the stated value.

[0018] Collagen composition

[0019] The biomaterials of the present invention include dehydrated collagen gels formed from collagen compositions, polymers or combinations of polymers, and additives or combinations of additives. As used herein, a collagen composition is any composition comprising collagen or any collagen derivative (such as partially hydrolyzed collagen or fully hydrolyzed collagen). According to the present invention, a collagen composition comprises (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen.

[0020] As used herein, collagen refers to collagen with a triple helix structure. Collagen can be acid-soluble collagen. Partially hydrolyzed collagen refers to a single chain of triple helix collagen. Partially hydrolyzed collagen may include colloids. Typically, as used herein, partially hydrolyzed collagen is a colloid. 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 a collagen hydrolysate. Partially hydrolyzed collagen and / or fully hydrolyzed collagen can be acid-soluble.

[0021] Collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen can each be independently derived from any animal source or product. Alternatively, collagen, partially hydrolyzed collagen, and / or fully hydrolyzed collagen can be prepared by an in vitro synthetic process. 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 (such as partially hydrolyzed collagen or fully hydrolyzed collagen) derived from a specific animal, fungal, or bacterial source or product refer to a collagen-containing component that, as part of a collagen composition, is extracted from an animal, fungal, or bacterial source or product and optionally has been further processed (e.g., hydrolyzed or purified) to produce collagen or collagen derivatives. For example, partially hydrolyzed collagen derived from an animal source or product is initially extracted from the animal source or product as part of a collagen composition and then obtained by hydrolysis of that collagen composition.

[0022] For example, collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen may be independently derived from seafood, bovine products, or swine products, preferably seafood or swine products. As used herein, collagen (or partially hydrolyzed collagen or fully hydrolyzed collagen) derived from marine, bovine, or swine sources or products may also be referred to as marine collagen, bovine collagen, or swine collagen (or partially hydrolyzed collagen or fully hydrolyzed collagen), respectively. Typically, at least one of the partially hydrolyzed collagen, collagen, and fully hydrolyzed collagen compositions is derived from seafood. 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, the collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen are not derived from bovine products. 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 different types of animal sources. For example, in one embodiment, the collagen composition comprises partially hydrolyzed marine collagen and porcine collagen.

[0023] Animal sources or products can be any part of an animal containing collagen. For example, seafood can be any part of a marine animal containing collagen. As used herein, marine animals can be any animal that is primarily or solely found in an aquatic environment, and can include animals found in both freshwater and marine environments. Marine animals can be a type of fish such as sea bass, snapper, brill, spotted catfish, catfish, black cod, cod, dabby, dogfish, eel, halibut, gar, haddock, halibut, mackerel, flounder, green cod, manta ray, salmon, sardine, ray, starfish, flounder, tilapia, or tuna. Marine animals can be invertebrates such as sea anemones, clams, corals, hydroids, jellyfish, mussels, oysters, scallops, sea cucumbers, sea slugs, conches, sea urchins, sponges, starfish, or worms. Marine animals can be arthropods, such as arachnids, crustaceans, insects, or myriapods.

[0024] Seafood can be freshwater fish products, saltwater fish products, invertebrate products, or arthropod products. In one embodiment, seafood is fish products, including both freshwater and saltwater fish. Fish products can be any part of a fish containing collagen. Typically, fish products include one or more of fish skin, scales, swim bladder, or joints and / or tendons; for example, it may include one or more of fish skin, scales, and / or swim bladder. All of these fish products contain collagen, although the swim bladder used in one preferred embodiment has a particularly high collagen content. In another preferred embodiment, fish products include fish skin. Fish products are more sustainable than similar collagen-containing products from other animals (such as bovine products) because the production of fish products requires less water and has a lower carbon footprint. In particular, fish skin is an readily available waste, thus its use has environmental benefits.

[0025] Bovine products can be any part of a bovine animal containing collagen. In one embodiment, the bovine product is bovine tendon. Bovine animals can be dairy cows, bison, buffalo, or antelopes. Typically, bovine products are dairy cow products. Pig products can be any part of a swine animal containing collagen. In one embodiment, the pig product is pigskin. Swine animals can be domestic pigs, wild pigs, or wild boars.

[0026] According to the present invention, the collagen composition comprises at least 30% by weight of partially hydrolyzed collagen. The presence of partially hydrolyzed collagen (such as gel) in the collagen composition improves the strength of biomaterials and any processed biomaterials produced from biomaterials. As used herein, the percentage by weight of collagen or collagen derivatives refers to the weight of collagen or collagen derivatives and is expressed as a weight percentage of all collagen or collagen-derived components in the collagen composition.

[0027] 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, no more than 95% by weight. The collagen composition may comprise 30% to 95% by weight, 50% to 95% by weight, 60% to 95% by weight, 70% to 95% by weight, 75% to 95% by weight, or 80% to 95% by weight of partially hydrolyzed collagen. Alternatively, the collagen composition may comprise 30% to 90% by weight, 50% to 90% by weight, 60% to 90% by weight, 70% to 90% by weight, 75% to 90% by weight, or 80% to 90% by weight of partially hydrolyzed collagen.

[0028] The collagen composition comprises collagen and / or fully hydrolyzed collagen. The collagen composition may include at least 1% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen. Typically, the collagen composition comprises at least 2% by weight, at least 3% by weight, at least 4% by weight, or at least 5% by weight, preferably at least 5% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen. The collagen composition may include at least 10% by weight, at least 15% by weight, at least 20% by weight, at least 30% by weight, at least 40% by weight, or at least 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen. The total collagen composition may include no more than 70% by weight, no more than 60% by weight, no more than 50% by weight, no more than 40% by weight, no more than 30% by weight, no more than 25% by weight, no more than 20% by weight, no more than 15% by weight, or no more than 10% by weight of collagen and fully hydrolyzed collagen. Typically, a collagen composition comprises 5% to 70% by weight, 5% to 50% by weight, 5% to 40% by weight, 5% to 30% by weight, 5% to 25% by weight, or 5% to 20% by weight of collagen and / or a mixture thereof. Alternatively, a collagen composition may comprise 10% to 70% by weight, 10% to 50% by weight, 10% to 40% by weight, 10% to 30% by weight, 10% to 25% by weight, or 10% to 20% by weight of collagen and / or a mixture thereof. Alternatively, a collagen composition may comprise 15% to 70% by weight, 15% to 50% by weight, 15% to 40% by weight, 15% to 30% by weight, 15% to 25% by weight, or 15% to 20% by weight of collagen and / or a mixture thereof.

[0029] The collagen composition may comprise at least 1% by weight of collagen and / or at least 1% by weight of fully hydrolyzed collagen. The collagen composition may comprise at least 2% by weight, at least 3% by weight, at least 4% by weight, or at least 5% by weight of one or both of collagen and fully hydrolyzed collagen. In one embodiment, the collagen composition comprises at least 5% by weight of collagen and / or at least 5% by weight of fully hydrolyzed collagen.

[0030] When the collagen composition contains collagen, the collagen composition may include 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 25% by weight of collagen, preferably at least 5% by weight or at least 10% by weight of collagen. Typically, when the collagen composition contains collagen, the collagen composition includes no more than 70% by weight of collagen, for example, no more than 60% by weight, no more than 50% by weight, no more than 40% by weight, no more than 30% by weight, no more than 25% by weight, no more than 20% by weight, no more than 15% by weight, or no more than 10% by weight of collagen. Preferably, the collagen composition includes no more than 50% by weight of collagen. The collagen composition may include no more than 30% by weight of collagen. Typically, the collagen composition includes 5% to 70% by weight, 5% to 50% by weight, 5% to 40% by weight, 5% to 30% by weight, 5% to 25% by weight, or 5% to 20% by weight of collagen. Preferably, the collagen composition comprises 5% to 50% by weight or 5% to 30% by weight of collagen. Alternatively, the collagen composition may comprise 10% to 70% by weight, 10% to 50% by weight, 10% to 40% by weight, 10% to 30% by weight, 10% to 25% by weight, or 10% to 20% by weight of collagen. Alternatively, the collagen composition may comprise 15% to 70% by weight, 15% to 50% by weight, 15% to 40% by weight, 15% to 30% by weight, 15% to 25% by weight, or 15% to 20% by weight of collagen. In one embodiment, the collagen composition does not contain collagen. Limiting the amount of collagen in a collagen composition allows the composition to contain more partially hydrolyzed collagen. The inventors have found that partially hydrolyzed collagen improves the tensile strength of biomaterials and increases the efficiency of the manufacturing process (particularly by reducing or eliminating the required neutralization and improving the handling of the product during manufacturing, especially by reducing the viscosity of the product and thus reducing bubble formation). Using less collagen also reduces the cost of the resulting biomaterials.

[0031] When a collagen composition contains fully hydrolyzed collagen, the collagen composition may include 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 25% by weight of fully hydrolyzed collagen. The presence of fully hydrolyzed collagen (e.g., collagen hydrolysate) in the collagen composition can improve the hardness, elasticity, ductility, and strength of biomaterials and any processed biomaterials produced from biomaterials. However, it is important to control the amount of fully hydrolyzed collagen in the collagen composition, as a large amount of fully hydrolyzed collagen weakens the collagen gel and the resulting biomaterial, and prevents the collagen gel from holding together well. When a collagen composition contains fully hydrolyzed collagen, the collagen composition typically includes no more than 50% by weight, no more than 40% by weight, no more than 30% by weight, no more than 25% by weight, no more than 20% by weight, no more than 15% by weight, or no more than 10% by weight of fully hydrolyzed collagen. Preferably, the collagen composition includes no more than 30% by weight or no more than 20% by weight of fully hydrolyzed collagen. Typically, the collagen composition comprises 5% to 50% by weight, 5% to 40% by weight, 5% to 30% by weight, 5% to 25% by weight, or 5% to 20% by weight of fully hydrolyzed collagen. Preferably, the collagen composition comprises 5% to 30% by weight or 5% to 20% by weight of fully hydrolyzed collagen. Alternatively, the collagen composition may comprise 10% to 50% by weight, 10% to 40% by weight, 10% to 30% by weight, 10% to 25% by weight, or 10% to 20% by weight of fully hydrolyzed collagen. Alternatively, the collagen composition may comprise 15% to 50% by weight, 15% to 40% by weight, 15% to 30% by weight, 15% to 25% by weight, or 15% to 20% by weight of fully hydrolyzed collagen. In one embodiment, the collagen composition does not contain fully hydrolyzed collagen.

[0032] In one embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of collagen and / or a mixture thereof. In one embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of collagen and / or a mixture thereof. In one embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of collagen and / or a mixture thereof. In a preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of collagen and / or a mixture thereof. In one preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of collagen and / or a mixture thereof. In another preferred embodiment, the collagen composition comprises (i) 70% to 90% by weight of partially hydrolyzed collagen and (ii) 10% to 30% by weight of collagen and / or a mixture thereof.

[0033] polymer

[0034] According to the present invention, in addition to the collagen composition, the collagen gel also includes a polymer.

[0035] The polymer may include a biopolymer or a bio-based polymer. In one embodiment, the polymer includes a biopolymer. In one embodiment, the polymer is a biopolymer. In one embodiment, the polymer includes a bio-based polymer. In one embodiment, the polymer is a bio-based polymer.

[0036] 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 plants, microorganisms, or animals. Biopolymers are generally biodegradable. Typically, biopolymers are polypeptides, proteins, polynucleotides, polysaccharides, isoprene polymers, gums, polyphenol polymers, lipids, fatty acid polymers, or pigments. In one embodiment, the biopolymer is a polysaccharide or gum. In one embodiment, the biopolymer is a polysaccharide.

[0037] As used herein, bio-based polymers are synthetic polymers in which at least a portion comprises materials derived from non-petrochemical sources (renewable raw materials). Typically, bio-based polymers are made from materials derived from non-petrochemical sources (renewable raw materials). Bio-based polymers are generally non-biodegradable.

[0038] The polymer can be water-soluble. Alternatively, the polymer can be insoluble in water.

[0039] The polymer may include one or a mixture of polymers selected from the following: cellulose, nanocellulose, lignin, starch, alginate, gum arabic, guar gum, gellan gum (including low-acyl gellan gum and high-acyl gellan gum), xanthan gum, carrageenan (including κ-carrageenan, ι-carrageenan, λ-carrageenan), polyhydroxyalkanoates (PHA), polyhydroxy fatty acids (PHF), bacterial cellulose, hyaluronic acid, hyaluronic acid, gel polysaccharides, pullulan, silk (including silk fibroin and silk fibers), elastin, chitin, Chitosan, casein, whey, albumin, polyisoprene (rubber), keratin, poultry feather fiber (PFF), mucin, pectin, agar, agarose, actin, fibrin, fibrinogen, suberin, keratin, melanin, cotton, collagen fiber, polylactic acid (PLA), wool, polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polypropylene terephthalate, polyurethane, polycarbonate, polyether ester copolymer, polyamide, polyesteramide, unsaturated polyester, epoxy resin, and phenolic resin. In one embodiment, the polymer comprises one or a mixture of polymers selected from the group consisting of: cellulose, nanocellulose, lignin, starch, alginate, gum arabic, guar gum, gellan gum (low-acyl and high-acyl), xanthan gum, carrageenan (including κ-carrageenan, ι-carrageenan, λ-carrageenan), polyhydroxyalkanoates (PHA), PHF, bacterial cellulose, hyaluronic acid, hyaluronic acid, gel polysaccharides, pullulan, silk (including silk fibroin and silk fibers), elastin, chitin, chitosan, casein, whey, albumin, polyisoprene (rubber), keratin, PFF, mucin, pectin, agar, agarose, actin, fibrin, fibrinogen, suberin, keratin, melanin, cotton, collagen fibers, and wool. As used herein, cellulose may be micron-cellulose or nanocellulose. Preferably, the cellulose is micron-cellulose.

[0040] Typically, polymers include 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 includes starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, cellulose, nanocellulose, collagen fibers, keratin, cotton, or mixtures thereof. In another embodiment, the polymer includes starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, guar gum, xanthan gum, cellulose, nanocellulose, keratin, cotton, or mixtures thereof. In yet another embodiment, the polymer includes starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, cellulose, nanocellulose, keratin, cotton, or mixtures thereof. Polymers may include starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, guar gum, xanthan gum, or mixtures thereof. The polymer may include starch, pectin, alginate, chitosan, agarose, gellan gum, gum arabic, or mixtures thereof. Alternatively, the polymer may include cellulose, nanocellulose, collagen fibers, keratin, cotton, or mixtures thereof. In one embodiment, the polymer includes cellulose, nanocellulose, keratin, cotton, or mixtures thereof. The polymer may include cellulose. In one embodiment, the polymer does not include collagen.

[0041] Water-insoluble polymers are typically present in collagen gels as reinforcing materials, such as fibers (including short or continuous fibers), films, sheets or plates, fragments, particles, fillers, whiskers, or spheres. For example, the polymer may be present in the collagen gel as fibers, typically fibers of biopolymers. In one embodiment, the polymer includes 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 includes cellulose fibers, nanocellulose fibers, cotton fibers, silk fibers, collagen fibers, wool fibers, keratin fibers, or mixtures thereof. In one embodiment, the polymer includes cellulose fibers, nanocellulose fibers, cotton fibers, silk fibers, wool fibers, keratin fibers, or mixtures thereof. In one embodiment, the polymer includes cellulose fibers, collagen fibers, keratin fibers, cotton fibers, or mixtures thereof. The polymer may include collagen fibers. In one embodiment, the polymer includes 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, the cellulose fibers may be micron-sized cellulose fibers or nanocellulose fibers. Preferably, the cellulose fibers are micron-sized cellulose fibers.

[0042] When the polymer includes collagen, the collagen is present in the form of a reinforcing material, such as collagen fibers. In one embodiment, the polymer includes no more than 15% of a collagen reinforcing material (e.g., collagen fibers). The polymer may include no more than 10% by weight or 5% by weight (e.g., no more than 2% by weight or 1% by weight) of collagen reinforcing material (e.g., collagen fibers) compared to the total weight of the collagen composition. In another embodiment, the polymer does not contain collagen.

[0043] Typically, collagen gels contain one polymer. However, gels can contain more than one polymer (i.e., a combination of polymers), such as two, three, or four different polymers. A gel may include two polymers. For example, a collagen gel may include starch and pectin, or cellulose fibers and wool fibers. In cases where a collagen gel includes more than one polymer, each polymer may be present in equal or unequal amounts.

[0044] The presence of polymers in collagen gels can be used to produce biomaterials. In particular, polymers or combinations of polymers improve the control over the mechanical properties of biomaterials and any processed biomaterials produced from them, compared to biomaterials that do not contain polymers. For example, polymers can allow for control over the stiffness of the material.

[0045] additive

[0046] According to the present invention, the collagen gel includes additives in addition to the polymer and collagen composition. Therefore, these additives differ from the polymer and collagen composition components of the collagen gel.

[0047] The additive is selected from anti-hydrolysis agents, foaming agents, thickeners, fillers, and flame retardants. The additive may be selected from anti-hydrolysis agents, foaming agents, thickeners, and flame retardants. Preferably, the additive is selected from anti-hydrolysis agents, foaming agents, and fillers. More preferably, the additive is selected from anti-hydrolysis agents, foaming agents, powdered fillers, and low-density fillers. In one embodiment, the additive is an anti-hydrolysis agent. In another embodiment, the additive is a foaming agent. In a further alternative embodiment, the additive is a filler, such as a powdered filler or a low-density filler. In one embodiment, the additive is not a filler.

[0048] Additives can be hydrolysis resistant agents. Antioxidants can also be used as hydrolysis resistant agents. The use of hydrolysis resistant agents generally improves the thermal stability of biomaterials and reduces material expansion under high humidity conditions. Suitable hydrolysis resistant agents include polycarbodiimides (e.g., Permutex). XR-13-906) and aliphatic carbodiimides (e.g., V1094, V1074, or LUBIO AS15). Preferably, the anti-hydrolysis agent is an aliphatic carbodiimide.

[0049] Additives can be foaming agents. Foaming agents are also called heat-generating agents. Surfactants can be used as foaming agents. Suitable foaming agents include N2O, sodium bicarbonate, sodium dodecyl sulfate (SDS), and ovalbumin. Preferably, the foaming agent is sodium bicarbonate or sodium dodecyl sulfate.

[0050] The additive may be a filler. Typically, the filler is a powdered filler or a low-density filler. In one embodiment, the filler is a powdered filler. In an alternative embodiment, the filler is a low-density filler. Fillers, especially powdered fillers and low-density fillers, generally improve the thermal stability of biomaterials and can reduce the elongation of biomaterials. Suitable powdered fillers include cork dust, colloidal silica, zeolite, calcium carbonate, and titanium dioxide. Preferably, the powdered filler is selected from cork dust, zeolite, and calcium carbonate. As used herein, a low-density filler refers to a filler with a density of less than about 5 g / cm³. 3 Typically less than about 3 g / cm³ 3 Or less than about 2.5 g / cm³ 3 The packing material. The density of low-density packing material can be approximately 2 g / cm³. 3 In one embodiment, the density of the low-density packing is less than about 1 g / cm³. 3 Suitable low-density fillers include glass bubbles (e.g., glass bubbles with a diameter of 60 µm and glass bubbles with a diameter of 30 µm, where the diameter represents the median diameter of the glass bubble swarm) and fused silica. Preferably, the low-density filler is glass bubbles. In one embodiment, the filler is not a polymer. In one embodiment, the filler is not cellulose (including micron-cellulose and nano-cellulose). Typically, the filler is not a fiber.

[0051] Additives can be flame retardants, such as zeolite.

[0052] Typically, collagen gels contain one additive. However, the gel may also contain more than one additive (i.e., a combination of additives), such as two or more additives (e.g., two, three, or four different additives), each independently selected from the additives described herein. More than one additive may include more than one additive from the same class of additives (e.g., the gel may include two foaming agents), and / or more than one additive from different classes of additives. A gel may include two additives, each independently selected from the additives described herein. For example, a collagen gel may contain sodium bicarbonate and SDS, or ovalbumin and SDS. In cases where a collagen gel includes more than one additive, each additive may be present in equal amounts or in different amounts.

[0053] Compared to biomaterials without additives, the use of additives or combinations of additives can improve the performance of biomaterials and / or any processed biomaterials produced from biomaterials. For example, additives can improve the thermal stability of materials and reduce material expansion under high humidity conditions.

[0054] Combination of polymers, collagen compositions and additives

[0055] The biomaterial of the present invention includes a dehydrated collagen gel, wherein the gel comprises (a) a collagen composition, (b) a polymer, and (c) an additive. Two or more polymers and / or two or more additives may be present in the gel.

[0056] Polymers (or polymer combinations) and collagen compositions can be linked by chemical bonds (such as chemical crosslinking bonds), by physical means (such as by physical winding or entanglement), or by a combination of both.

[0057] In one embodiment, the collagen composition and one or more polymers are linked by chemical cross-linking bonds. Alternatively, the collagen composition and one or more polymers may be physically intertwined.

[0058] The combination of a water-soluble polymer and a collagen composition may be referred to herein as a polymer blend. In a polymer blend, the polymer and the collagen composition are homogeneous, i.e., forming a single phase. In one embodiment, a collagen gel comprises a polymer blend that includes a collagen composition and a polymer. In another embodiment, a collagen gel comprises a polymer blend that includes a collagen composition and a polymer, wherein the collagen composition and the polymer are chemically crosslinked.

[0059] A combination of a water-insoluble polymer and a collagen composition may be referred to herein as a composite material. In a composite material, the polymer and the collagen composition are heterogeneous, i.e., in different phases. In such a composite material, the polymer is typically present as a reinforcing material, for example as fibers (including short or continuous fibers), films, sheets or plates, fragments, particles, fillers, whiskers, or spheres. In one embodiment, a collagen gel comprises a composite material comprising a collagen composition and a polymer. In another embodiment, a collagen gel comprises a composite material comprising a collagen composition and a polymer, wherein the collagen composition and the polymer are physically wound and / or chemically crosslinked.

[0060] The nature of the interaction between the collagen composition and one or more polymers will depend on factors such as the type of polymer used, the type of crosslinking agent, the crosslinking conditions, and the solubility of the polymer. For example, in cases where the polymer has functional groups that can react with the crosslinking agent, chemically crosslinked materials can be formed, thereby creating chemical crosslinks between the polymer and the collagen composition. If the polymer does not contain any functional groups suitable for crosslinking, the polymer and collagen composition will typically be physically intertwined. Those skilled in the art will readily understand how to achieve chemical crosslinking and / or physical intertwining of a particular polymer.

[0061] If a collagen gel uses more than one type of polymer, the polymers can be linked to the collagen composition in the same or different ways. For example, a collagen gel may include a first polymer that is chemically cross-linked with the collagen composition and a second polymer that is physically entangled with the collagen composition and the first polymer. Alternatively, both the first and second polymers may be chemically cross-linked or physically entangled with the collagen composition.

[0062] Collagen gels typically comprise at least 1% by weight of a polymer compared to the weight of the collagen composition. Collagen gels may comprise at least 2%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, or at least 50% by weight of a polymer compared to the weight of the collagen composition. Preferably, the collagen gel comprises at least 2% or at least 5% by weight of a polymer compared to the weight of the collagen composition.

[0063] The collagen gel may comprise no more than 50% by weight, no more than 40% by weight, no more than 30% by weight, no more than 25% by weight, no more than 20% by weight, no more than 15% by weight, no more than 10% by weight, no more than 5% by weight, no more than 2% by weight, or no more than 1% by weight of polymer compared to the weight of the collagen composition. Preferably, the collagen gel comprises no more than 25% by weight or no more than 20% by weight of polymer compared to the weight of the collagen composition.

[0064] The collagen gel may comprise, by weight, 1% to 50% of polymer, 1% to 40% of polymer, 1% to 30% of polymer, 1% to 25% of polymer, 1% to 20% of polymer, 1% to 15% of polymer, 1% to 10% of polymer, or 1% to 5% of polymer, compared to the weight of the collagen composition. Alternatively, the collagen gel may comprise, by weight, 2% to 50% of polymer, 2% to 40% of polymer, 2% to 30% of polymer, 2% to 25% of polymer, 2% to 20% of polymer, 2% to 15% of polymer, 2% to 10% of polymer, or 2% to 5% of polymer, compared to the weight of the collagen composition. Alternatively, the collagen gel may comprise, by weight, 5% to 50% of polymer, 5% to 40% of polymer, 5% to 30% of polymer, 5% to 25% of polymer, 5% to 20% of polymer, 5% to 15% of polymer, or 5% to 10% of polymer, compared to the weight of the collagen composition. Preferably, the collagen gel comprises a polymer comprising 1% to 25% by weight, 1% to 20% by weight, 2% to 20% by weight, 2% to 15% by weight, or 2% to 10% by weight, relative to the weight of the collagen composition.

[0065] When a collagen gel comprises more than one polymer, the collagen gel may include each polymer within the aforementioned weight range. Alternatively, the collagen gel may include the total amount of polymers within the aforementioned weight range.

[0066] In one embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 30% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 70% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 10% by weight of polymer compared to the weight of the collagen composition.

[0067] In one embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 50% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 50% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 10% by weight of polymer compared to the weight of the collagen composition.

[0068] In one embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition. In another embodiment, the collagen composition comprises (i) 60% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 40% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 10% by weight of polymer compared to the weight of the collagen composition.

[0069] In one preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition. In another preferred embodiment, the collagen composition comprises (i) 70% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 10% by weight of polymer compared to the weight of the collagen composition.

[0070] In one preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition. In another preferred embodiment, the collagen composition comprises (i) 80% to 95% by weight of partially hydrolyzed collagen and (ii) 5% to 20% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 10% by weight of polymer compared to the weight of the collagen composition.

[0071] In one preferred embodiment, the collagen composition comprises (i) 70% to 90% by weight of partially hydrolyzed collagen and (ii) 10% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 25% by weight of polymer compared to the weight of the collagen composition. In another preferred embodiment, the collagen composition comprises (i) 70% to 90% by weight of partially hydrolyzed collagen and (ii) 10% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 1% to 20% by weight of polymer compared to the weight of the collagen composition. In yet another preferred embodiment, the collagen composition comprises (i) 70% to 90% by weight of partially hydrolyzed collagen and (ii) 10% to 30% by weight of one or a mixture of collagen and / or fully hydrolyzed collagen, and the collagen gel comprises 2% to 20% by weight of polymer compared to the weight of the collagen composition. In one preferred embodiment, the collagen composition comprises (i) 70% to 90% by weight of partially hydrolyzed collagen and (ii) 10% to 30% by weight of collagen and / or a mixture of fully hydrolyzed collagen, and the collagen gel comprises 2% to 15% by weight of polymer compared to the weight of the collagen composition.

[0072] As used herein and unless otherwise stated, references to additives by weight (%) refer to the percentage of the additive's weight relative to the weight of the protein in the collagen composition (i.e., the weight of the collagen or collagen derivative component (including partially and fully hydrolyzed collagen) in the collagen composition). References to the amount of additive indicated by M refer to the number of moles of additive contained per liter of solution containing the collagen composition (i.e., a solution of collagen or collagen derivative component (including partially and fully hydrolyzed collagen)).

[0073] When the collagen gel contains an anti-hydrolysis agent, the content of the anti-hydrolysis agent is typically at least 0.1 wt%, for example, at least 0.5 wt%, at least 1.0 wt%, at least 1.5 wt%, at least 2.0 wt%, or at least 2.5 wt%. Preferably, the content of the anti-hydrolysis agent is at least 0.5 wt%, at least 1.0 wt%, or at least 1.5 wt%, for example, at least 0.5 wt%. The content of the anti-hydrolysis agent is typically no more than 5 wt%, for example, no more than 4 wt%, no more than 3 wt%, or no more than 2 wt%. Preferably, the content of the anti-hydrolysis agent is no more than 3 wt% or no more than 2 wt%, for example, no more than 3 wt%. The content of the anti-hydrolysis agent is typically from 0.1 wt% to 5 wt%, for example, from 0.5 wt% to 5 wt%, from 0.5 wt% to 4 wt%, or from 0.5 wt% to 3 wt%. Preferably, the content of the anti-hydrolysis agent is from 0.5 wt% to 3 wt%.

[0074] The content of a foaming agent in a collagen gel typically depends on the foaming agent used. When the collagen gel includes a foaming agent, and the foaming agent is N2O, the N2O content is typically at least 100% by weight, for example, at least 120% by weight, at least 140% by weight, at least 160% by weight, or at least 180% by weight. Preferably, the N2O content is at least 120% by weight or at least 140% by weight. The N2O content typically does not exceed 200% by weight, for example, not exceeding 180% by weight or not exceeding 160% by weight. Preferably, the N2O content does not exceed 180% by weight. The N2O content is typically from 100% by weight to 200% by weight, for example, from 120% by weight to 180% by weight or from 140% by weight to 180% by weight. Preferably, the N2O content is from 140% by weight to 180% by weight. When the collagen gel contains a foaming agent, and the foaming agent is sodium bicarbonate, the content of sodium bicarbonate is typically at least 5% by weight, for example, at least 10% by weight, at least 20% by weight, at least 25% by weight, or at least 50% by weight. Preferably, the content of sodium bicarbonate is at least 10% by weight or at least 20% by weight. The content of sodium bicarbonate is typically no more than 70% by weight, for example, no more than 60% by weight, no more than 50% by weight, or no more than 40% by weight. Preferably, the content of sodium bicarbonate is no more than 60% by weight or no more than 50% by weight. The content of sodium bicarbonate is typically from 5% by weight to 70% by weight. In one embodiment, the content of sodium bicarbonate is from 5% by weight to 50% by weight, for example, from 10% by weight to 50% by weight or from 10% by weight to 40% by weight. In one embodiment, the content of sodium bicarbonate is from 50% by weight to 70% by weight. When the collagen gel contains a foaming agent, and the foaming agent is ovalbumin, the content of ovalbumin is typically at least 1% by weight, for example, at least 2% by weight, at least 5% by weight, at least 8% by weight, or at least 10% by weight. Preferably, the ovalbumin content is at least 2% by weight or at least 5% by weight. The ovalbumin content typically does not exceed 20% by weight, for example, not more than 15% by weight, not more than 12% by weight, or not more than 10% by weight. Preferably, the ovalbumin content does not exceed 15% by weight or not more than 12% by weight. The ovalbumin content typically ranges from 1% by weight to 20% by weight, for example, 2% by weight to 15% by weight, 5% by weight to 15% by weight, or 8% by weight to 12% by weight. Preferably, the ovalbumin content is from 5% by weight to 15% by weight or from 8% by weight to 12% by weight. When the collagen gel includes a foaming agent and the foaming agent is sodium lauryl sulfate (SDS), the SDS content is typically at least 0.1 mM, for example, at least 0.5 mM, or at least 0.75 mM, or at least 1.0 mM, or at least 1.25 mM, or at least 1.5 mM. Preferably, the SDS content is at least 0.5 mM or at least 0.75 mM. The SDS content is usually no more than 5 mM, for example no more than 4 mM, or no more than 3 mM or no more than 2.5 mM.Preferably, the SDS content does not exceed 3 mM or 2.5 mM. The SDS content is typically from 0.1 mM to 5 mM, for example, 0.5 mM to 2.5 mM, or 0.75 mM to 2.5 mM or 1.0 mM to 2.5 mM. Preferably, the SDS content is from 0.5 mM to 2.5 mM or 0.75 mM to 2.5 mM.

[0075] When collagen gels include fillers (e.g., powdered fillers or low-density fillers), the filler content is typically at least 0.1 wt%, for example, at least 0.5 wt%, at least 1.0 wt%, at least 2 wt%, or at least 5 wt%. Preferably, the filler content is at least 0.5 wt% or at least 1.0 wt%. The filler content typically does not exceed 20 wt%, for example, not more than 18 wt%, not more than 15 wt%, not more than 12 wt%, or not more than 10 wt%. Preferably, the filler content does not exceed 15 wt% or not more than 12 wt%. The filler content is typically from 0.1 wt% to 20 wt%, for example, from 0.5 wt% to 18 wt%, or from 1.0 wt% to 15 wt%, or from 2 wt% to 12 wt%. Preferably, the filler content is from 0.5 wt% to 18 wt%, or from 1.0 wt% to 15 wt%.

[0076] Extracted collagen composition

[0077] In one embodiment, the collagen gel can be formed from a collagen composition extracted from animal products, particularly seafood. As used herein, a collagen composition extracted from animal products (e.g., seafood) can be described as an extracted collagen composition. As used herein, a collagen composition extracted from seafood can be described as a marine collagen composition. The seafood mentioned above has been described.

[0078] Advantageously, collagen compositions extracted from seafood can be efficiently utilized to create biomaterials that are well-suited for further processing steps to form leather-like biomaterials. Previously known methods for producing biomaterials from collagen do not use seafood as a collagen source and often require lengthy and complex extraction steps to provide collagen in a form suitable for further processing. Cultured collagen has also been used, but this is inefficient and the process is not easily scalable. Another disadvantage of cultured collagen is that it does not provide an endogenous mixture of natural collagen proteins.

[0079] Collagen in the extracted collagen composition can be extracted using acid (i.e., acid-soluble collagen) or pepsin (i.e., pepsin-soluble collagen). 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 a greater amount of partially and / or fully hydrolyzed collagen. For example, partially hydrolyzed 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 defined herein includes an extracted collagen composition, such as a collagen composition extracted from seafood.

[0080] The presence of partially and / or fully hydrolyzed collagen in the extracted collagen composition can be used to produce biomaterials. In particular, partially and / or fully hydrolyzed collagen can improve 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 them. Specifically, the presence of partially hydrolyzed collagen can result in biomaterials and / or processed biomaterials that are softer and more elastic compared to biomaterials without partially hydrolyzed collagen.

[0081] Extracted collagen compositions typically comprise collagen and optionally partially and / or fully hydrolyzed collagen. Generally, extracted collagen compositions comprise at least one of acid-soluble collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen. As used herein, acid-soluble collagen is collagen that can be extracted using acid. Extracted collagen compositions may contain at least 20% by weight, at least 30% by weight, or at least 40% by weight of acid-soluble collagen. For example, extracted collagen compositions may contain from 20% by weight to 50% by weight of acid-soluble collagen. Extracted collagen compositions may 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, extracted collagen compositions may contain from 1% by weight to 40% by weight of partially hydrolyzed collagen. Extracted collagen compositions may 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 fully hydrolyzed collagen. For example, the extracted collagen composition may contain 1% to 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 another 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 of acid-soluble collagen provided above can also be applied to collagen.

[0082] The extracted collagen composition can be an endogenous composition, meaning it contains collagen, collagen derivatives (such as partially and / or fully hydrolyzed collagen), and other components (such as naturally occurring impurities) because they are naturally present in seafood. For example, collagen will typically have intact terminal peptide regions. This can make products formed from the extracted collagen composition more appealing to certain consumer groups. Advantageously, collagen compositions extracted from seafood can be used in their endogenous form without requiring complex processing. Furthermore, the endogenous composition can contain collagen derivatives that can improve the efficiency of biomaterial manufacturing and advantageously influence the performance of any processed biomaterial produced from it.

[0083] Manufacturing method

[0084] This invention also relates to methods for manufacturing the biomaterials described herein. Typically, methods for producing biomaterials include:

[0085] a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and

[0086] b) Dehydrate the collagen gel to form biomaterials.

[0087] The collagen composition can be any collagen composition as defined herein. In one embodiment, the present invention provides a method for producing biomaterials, the method comprising:

[0088] a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and

[0089] b) Dehydrating collagen gel to form biomaterials;

[0090] The collagen composition comprises (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen, wherein

[0091] The collagen composition comprises at least 30% by weight of partially hydrolyzed collagen, and

[0092] The additives are selected from anti-hydrolysis agents, foaming agents, thickeners, fillers and flame retardants.

[0093] The polymer can be any polymer or combination of polymers as defined herein. The additive can be any additive or combination of additives as defined herein.

[0094] In one embodiment, the forming step (a) includes contacting the collagen composition, polymer, and additives 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 and / or additives are added to the crosslinkable collagen mixture prior to any crosslinking. However, the polymer and / or additives may be added to a partially crosslinked crosslinkable collagen mixture, i.e., immediately after some crosslinking has occurred.

[0095] When an additive, acting as a foaming agent, is to be added to a biomaterial, step (a) may include agitating the crosslinkable collagen mixture (e.g., using a mechanical beater, stirrer, or mixer) or oscillating the crosslinkable collagen mixture. Typically, agitation and / or oscillation are performed immediately after one or more crosslinking agents have been added to the mixture of the collagen composition, polymer, and additive. When the foaming agent is N₂O, oscillation is typically performed in a closed container after the addition of pressurized N₂O. Alternatively, step (a) may include contacting an acid (e.g., hydrochloric acid) with the collagen composition, polymer, additive, and crosslinking agent. Typically, the acid is added to the mixture of the collagen composition, polymer, and additive first, followed by the crosslinking agent. This is typically used when the foaming agent is an alkaline reagent (e.g., sodium bicarbonate) that reacts with the acid to produce bubbles.

[0096] In some embodiments, the crosslinking forms crosslinking bonds between the collagen composition and the polymer. Alternatively or additionally, the crosslinking forms crosslinking bonds within the collagen composition and around the polymer, causing the collagen composition and the polymer to be physically entangled.

[0097] The formation step may include adding a fatliquoring component and / or dye or pigment to a collagen composition, a crosslinkable collagen mixture, a polymer, or an additive. The formation step is further described below. In one embodiment, the present invention provides a method for producing the biomaterial of the present invention, the method comprising:

[0098] a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and

[0099] b) Dehydrating collagen gel to form biomaterials;

[0100] The formation step includes adding fatliquoring components and / or dyes or pigments to the collagen composition.

[0101] In one embodiment, the present invention provides a method for producing the biomaterial of the present invention, the method comprising:

[0102] a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and

[0103] b) Dehydrating collagen gel to form biomaterials;

[0104] The forming step includes contacting the collagen composition, polymer, and additives 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 fat-adding components and / or dyes or pigments to the collagen composition, crosslinkable collagen mixture, polymer, or additives.

[0105] This article also discloses a method for producing biomaterials, the method comprising:

[0106] a) Extraction of collagen compositions from animal products, preferably seafood; and

[0107] b) Forming a collagen gel comprising a collagen composition, a polymer, and additives, and dehydrating the collagen gel to form a biomaterial; wherein the collagen composition comprises collagen and optionally partially and / or fully hydrolyzed collagen.

[0108] Extraction step (a) may further include adding partially hydrolyzed collagen 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, wherein the collagen composition comprises 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, wherein the collagen composition comprises at least 30% by weight of partially hydrolyzed collagen.

[0109] Any of the methods described herein may also include processing the biomaterial to form a processed biomaterial. The processed biomaterial may be a leathery biomaterial. One or more processing steps may include one or more of drying, dyeing, fatliquoring, finishing, and coating of the biomaterial.

[0110] Any of the methods described herein may also include adding a textile pad to a collagen gel, biomaterial, or processed biomaterial. Suitable textiles known to those skilled in the art include cotton, wool, silk, leather, goatskin, flax, jute, hemp, felt, linen, fiberglass cloth, nylon, polyester, Tencel, rayon, Kevlar, viscose, spandex, acrylic, polyamide, and mixtures thereof. Additional suitable textiles include bamboo, fur, ramie, polyurethane, polypropylene, cellulose acetate, rubber, neoprene, expanded polytetrafluoroethylene (e.g., Gore-Tex), polyethylene (e.g., Tyvek), m-phenylenediamine / isophthaloyl chloride copolymer (e.g., Nomex aramid), and mixtures thereof. Textile blends and / or recycled textiles are also included. Adding a textile pad may involve bonding a textile pad layer to a biomaterial or processed biomaterial. For example, textile padding can be added by laminating an adhesive layer onto a biomaterial / processed biomaterial and / or a textile padding layer using a coating method. Suitable coating methods are known to those skilled in the art, including spraying, brushing, dagger-roll coating, roll-to-roll coating, reverse roll coating, curtain coating, and slot coating. The lamination process can use lamination rollers (as part of a roll-to-roll process) or lamination lines (where lamination is performed using pressure and / or heat to bond the padding layer to the collagen gel / biomaterial / processed biomaterial, typically via an adhesive; this adhesive itself may be on the roller). Alternatively, the textile padding layer can be placed on or within a crosslinkable collagen mixture before crosslinking (or on or within a partially crosslinked crosslinkable collagen mixture), for example, by partially or completely immersing the textile in the crosslinkable collagen mixture and then bonding it to the biomaterial via crosslinking bonds during the (remaining) crosslinking step. Textile padding can also be added by casting the biomaterial or processed biomaterial onto the textile padding layer.

[0111] In one embodiment, the extraction step (a) includes washing the animal product, such as seafood, with an alkaline solution, optionally further washing the seafood with a defatting agent, contacting the seafood with an acidic solution of pH 4 to 5, and obtaining the extracted collagen composition from the acidic solution; the formation step (b) includes contacting the extracted collagen composition, polymer, and additives 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; and optionally, the method further includes a processing step (c) following the formation step (b) to form processed biomaterial, wherein the processing step includes fatliquoring the biomaterial, staining the fatliquored biomaterial, drying the stained and fatliquored biomaterial, and mechanically processing the dried biomaterial.

[0112] This article also describes a biomaterial that can be obtained by methods as described herein. The biomaterial may be a leather-like biomaterial.

[0113] (a) Extraction

[0114] The following sections describe the extraction of collagen compositions from animal products. References to animal products herein may be considered as references to specific animal products (e.g., seafood) from which collagen is extracted.

[0115] Collagen can be extracted from animal products, such as seafood, by contacting the animal products with an acidic solution. The acidic solution can be any solution with a pH less than 7. Typically, the acidic solution may have a pH of 3 to 6, preferably 4 to 6. In a preferred embodiment, the acidic solution has a pH of 4 to 5. The acidic solution may comprise any weak acid or diluted strong acid with a suitable 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.

[0116] Animal products may be contacted with an acidic solution for at least 6 hours, at least 12 hours, or at least 24 hours. Typically, animal products are contacted with an acidic solution for approximately 12 hours. Animal products and the acidic solution may be contacted at temperatures below 30°C, below 20°C, or below 10°C. Typically, animal products and the acidic solution are contacted at a temperature of approximately 4°C. After contacting the animal products with the acidic solution, the extracted collagen composition can be separated from the solution. The extracted collagen composition may be freeze-dried (lyophilized).

[0117] The extraction step may also include washing the animal products with an alkaline solution before contacting them with an acidic solution. The animal products may be washed with an alkaline solution more than once. For example, they may be washed twice with an alkaline solution. The alkaline solution can be used to remove non-collagenous proteins from the animal products and to break them down. The alkaline solution can 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 sodium hydroxide solution.

[0118] The extraction step may also include washing the animal product with a defatting agent before contacting it with an acidic solution. The defatting agent is used to remove fat from the product. Typically, the defatting agent is an alcohol solution, an organic solvent (such as chloroform, petroleum ether, or n-hexane), or supercritical CO2. In a preferred embodiment, the defatting agent is an alcohol solution. The alcohol content in the water of the alcohol solution is less than 70% v / v, less than 50% v / v, or less than 30% v / v. Typically, the alcohol content in the water of the alcohol solution is between 5% v / v and 20% v / v. Using a solution of alcohol in water instead of pure alcohol prevents dehydration of the animal product, which reduces the extraction efficiency of the collagen composition. The alcohol can be methanol, ethanol, propanol, propanol (isopropanol), butanol, or butanol. In one embodiment, the alcohol solution is a solution of isopropanol.

[0119] In cases where animal products are washed with both an alkaline solution and a degreasing agent before being exposed to an acidic solution, the alkaline solution washing may be performed before or after the degreasing agent washing. Typically, animal products are washed with an alkaline solution before the degreasing agent washing. In one embodiment, the animal products are washed with an alkaline solution before the alcohol solution washing.

[0120] Animal products can be washed with water before and / or after each part of the extraction process. For example, animal products can be washed before contact with an alkaline solution, between contact with an alkaline solution and a degreasing agent, and between contact with a degreasing agent and an acidic solution.

[0121] The extraction method described herein does not require further processing steps such as enzymatic digestion or purification of the collagen composition. Compared to methods that require such steps, this reduces the time and resources required to perform the extraction. It also preserves the collagen protein in an undigested form, eliminating the need to break protein chains.

[0122] (b) Formation

[0123] The collagen composition may be or may include an extracted collagen composition, which is formed into a collagen gel with a polymer or polymer combination and an additive or additive combination, and the collagen gel is dehydrated to form a biomaterial.

[0124] Typically, the collagen composition is first provided in a suitable solution for forming the biomaterial. For example, the collagen composition may be diluted in water or a buffer solution, or a lyophilized collagen composition may be dissolved in water or a buffer solution. Suitable concentrations are from 1 mg / mL to 200 mg / mL, for example from 10 mg / mL to 100 mg / mL, of collagen proteins (including collagen, acid-soluble collagen, partially hydrolyzed collagen, and fully hydrolyzed collagen).

[0125] Advantageously, the collagen composition used in this invention may comprise an extracted collagen composition, which is generally highly soluble in aqueous solutions at pH 5 to 8 (e.g., pH 6 to 8, pH 6 to 7, or approximately pH 7). This means that a small amount of aqueous solvent (such as water) can be used without the need to add a large amount of acid to lower the pH and dissolve the collagen composition. In one embodiment, the solubility of the collagen composition at pH 5 to 8 and at a temperature of 25°C is at least 20 mg / mL, at least 30 mg, at least 40 mg, or at least 50 mg of collagen composition per mL of aqueous solvent. Preferably, the solubility of the collagen composition at pH 6 to 7 and at a temperature of 25°C is at least 20 mg / mL, at least 30 mg, at least 40 mg, or at least 50 mg of collagen composition per mL of aqueous solvent. For collagen compositions containing collagen, it may be necessary to add an acidic solvent to achieve dissolution. Any suitable weak acid or diluted strong acid, such as an aqueous solution of acetic acid, formic acid, or hydrochloric acid, can be used. Typically, collagen is dissolved separately in an acidic solvent along with partially hydrolyzed and / or fully hydrolyzed collagen. The collagen solution is then added to the solution of partially hydrolyzed and / or fully hydrolyzed collagen to obtain a combined solution of the collagen composition at a pH of 5 to 8 (preferably, pH 6 to 8 or 6 to 7). Collagen can be dissolved in aqueous solutions with a pH 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 approximately 2. However, in collagen compositions containing small amounts of collagen, only a small amount of acidic solution is required. Furthermore, the entire collagen composition is generally still soluble in solutions with pH values ​​between 5 and 8. Physical agitation such as stirring, mixing, and sonication can also be used to aid dissolution. Increased temperatures (i.e., temperatures above approximately 25°C) can also be used to aid dissolution. Because a smaller amount of solvent is required, it is advantageous to use a highly soluble collagen composition in this step, and the solvent is easier to remove during dehydration.

[0126] This contrasts with collagen blends, which are typically used in known processes to produce biomaterials and contain only or primarily collagen. Such collagen blends are generally insoluble at pH values ​​between 5 and 8 and require much lower pH values ​​to dissolve, such as below 5. Using a collagen composition that does not require highly acidic conditions for dissolution means that the solution can be easily neutralized later during gel formation. This improves manufacturing efficiency and further reduces the total amount of solvent required.

[0127] Typically, polymers (or combinations of polymers) and additives (or combinations of additives) are added to a solution of the collagen composition. When the polymers and / or additives are water-soluble, the same techniques described above can be used to dissolve the polymers and / or additives together with the collagen composition in the solution. Alternatively, the polymers and / or additives can be dissolved separately from the collagen composition, and then the solutions of the polymers and / or additives and the collagen composition can be combined. The polymers and additives can be dissolved in the same separate solutions or in different solutions. When the polymers and / or additives are insoluble in water, they are typically added to the solution of the collagen composition immediately after the collagen composition has been completely dissolved.

[0128] As part of the formation step, the collagen composition may be or may include an extracted collagen composition and optionally a polymer and / or additives (where the polymer and / or additives are capable of forming chemical crosslinks). The collagen composition may be crosslinked using any suitable protein crosslinking method known in the art. Typically, the collagen composition, one or more polymers, and one or more additives are contacted with one or more crosslinking agents to form a crosslinkable collagen mixture. The one or more crosslinking agents may be any molecule having two, three, or more functional reactive groups (capable of forming crosslinks between collagen molecules). Alternatively, the one or more crosslinking agents may be molecules that can be used to initiate a photo-initiated crosslinking process. The crosslinking agent may be an enzyme. Typically, the one or more crosslinking agents are one or more reagents selected from the group consisting of alcohols, aldehydes, amines, azides, carboxylic acids, carbodiimides, chromium salts, epoxides, acylhydrazides, isocyanates, mercapto groups, N-hydroxysuccinimides, imino esters, maleimides, haloacetyl groups, pyridyl disulfides, aryl azides, diaziridine, aglycones, and Staudinger linkers. Typically, one or more cross-linking agents may include glutaraldehyde, transglutaminase, carbodiimide, hydrazide, or genipin. For example, one or more cross-linking agents may include glutaraldehyde and / or transglutaminase. Suitable amounts of cross-linking agents are known to those skilled in the art. Typically, based on the total weight of the collagen composition, 0.1 wt% to 40 wt%, for example, 1 wt% to 10 wt%, of the cross-linking agent can be used. When glutaraldehyde is used as the cross-linking agent, the amount of glutaraldehyde in the cross-linkable composition may be, for example, 0.5 wt% to 10 wt%, for example, 1 wt% to 5 wt%. Alternatively, when the cross-linking agent is an enzyme, the amount of enzyme is typically 0.1 U to 70 U per gram of collagen composition, for example, 0.1 U to 40 U, ​​1 U / g to 60 U / g, 10 U / g to 60 U / g, 20 U / g to 60 U / g, or 30 U / g to 60 U / g. The amount of cross-linking agent can be from 1 U / g to 10 U / g. When transglutaminase is used as the cross-linking agent, the amount of transglutaminase can be, for example, from 0.5 U / g to 10 U / g, or, for example, from 1 U / g to 5 U / g. Alternatively, the amount of transglutaminase can be from 5 U / g to 60 U / g, for example, from 10 U / g to 60 U / g or from 20 U / g to 60 U / g.

[0129] Crosslinkable collagen blends may also contain dyes or pigments. Therefore, in one embodiment, a dye or pigment is added to the collagen composition, the crosslinkable collagen blend, the polymer, or the additive. For example, the dye or pigment may be a water-based dye or pigment, an alcohol-based dye or pigment, an acid dye, a direct dye, a mordant dye, or a basic dye. In one embodiment, the dye is a water-based dye or a water-based pigment. Using a dye in this step helps ensure that the resulting biomaterial is uniformly stained across its thickness.

[0130] Crosslinkable collagen blends may also contain fatliquoring components, such as fatliquoring emulsions. Therefore, in one embodiment, a fatliquoring component is added to the collagen composition, crosslinkable collagen blend, polymer, or additive. Fatliquoring emulsions may include salts of fats, such as sulfonates, sulfites, and / or phosphates of triglycerides. Using a fatliquoring emulsion at this step improves the penetration depth, speed, and uniformity of the fatliquoring agent into the biomaterial compared to fatliquoring after gel formation, while still providing beneficial softening and waterproofing properties. Fatliquoring is further described in the description of the processing steps.

[0131] Crosslinkable collagen mixtures may include one or more other additional components, such as one or more plasticizers. Plasticizers help make the resulting biomaterial soft and flexible. Suitable plasticizers are known to those skilled in the art. In one embodiment, the plasticizer is glycerol, and the crosslinkable collagen mixture includes glycerol. Based on the weight of the collagen composition, the plasticizer may be used in amounts, for example, from 5 wt% to 50 wt%. When glycerol (also known as glycerol) is used as the plasticizer, the amount of glycerol in the crosslinkable composition may be, for example, from 10 wt% to 40 wt%, such as 20 wt% to 40 wt%.

[0132] One or more defoamers can also be added to the crosslinkable collagen mixture, such that the collagen composition (and the crosslinkable collagen mixture) further includes one or more defoamers. Defoamers can also be referred to as defoaming agents. Defoamers remove air bubbles or foam that form within the composition, which improves the handling of the composition and helps form a more homogeneous biomaterial. Typically, physical agitation, such as stirring or agitation, is used in conjunction with defoamers to ensure complete elimination of air bubbles and foam. In one embodiment, physical agitation may be applied to the collagen composition and / or crosslinkable collagen mixture including the defoamer 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 defoamers compared to mixtures containing only collagen. For example, the inventors have attempted to reproduce Example 2 of EP3205668, which describes leather manufactured using a biomanufacturing technique from bovine collagen formation. It was found that the crosslinkable collagen mixture was very thick and contained a large number of air bubbles that were not easily removed even at pH 2 by conventional methods such as ultrasonic treatment. It is believed that the presence of partially hydrolyzed collagen in the collagen used in this invention produces a thinner and non-viscous mixture, which helps to remove foam using defoamers.

[0133] Suitable defoamers are known to those skilled in the art. In one embodiment, the defoamer is a food-grade defoamer. The defoamer may be a silicone-based emulsion, a polypropylene glycol composition, or a copolymer of ethylene oxide (EO) and propylene oxide (PO). In one embodiment, the defoamer is a silicone-based emulsion. The defoamer may be used, for example, in amounts from 0.001 wt% to 5 wt%, where wt% refers to the weight of the active ingredient of the defoamer relative to the weight of the total solution. Typically, the amount of defoamer added to the solution will depend on the amount of foam generated during formation, which may be affected by the various molecular weights of collagen extracted from different sources. The defoamer 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, defoamers are used in amounts ranging from 0.001 wt% to 5 wt%, 0.01 wt% to 3 wt%, 0.1 wt% to 3 wt%, 0.1 wt% to 2 wt%, 0.1 wt% to 1 wt%, or 0.1 wt% to 0.5 wt%. Preferably, defoamers are used in amounts not exceeding 1 wt%. For example, defoamers can be used in amounts ranging from 0.001 wt% to 1 wt%, 0.01 wt% to 1 wt%, or 0.1 wt% to 1 wt%. When a silicone-based emulsion is used as the defoamer, the amount of silicone-based emulsion in the crosslinkable composition can be, for example, 0.001 wt% to 5 wt%, or, for example, 0.1 wt% to 2 wt%.

[0134] Defoamers can also improve the properties of biomaterials and any processed biomaterials produced from biomaterials. In particular, it has been found that low concentrations of defoamers (e.g., 0.1 wt% to 3 wt%) increase the tensile strength of the resulting biomaterials.

[0135] Typically, crosslinkable collagen mixtures have a pH of 6 to 8, preferably about 7, before crosslinking. If the crosslinkable collagen mixture has a pH outside this range, a suitable amount of acid or base can be added to achieve the desired pH. As mentioned above, the collagen compositions used in this invention are advantageously highly soluble in aqueous solutions at pH values ​​of 5 to 8 (preferably 6 to 8 or 6 to 7). Therefore, in one embodiment, the formation step does not require neutralization to obtain a crosslinkable mixture with a pH of 6 to 8. In one embodiment, the formation step involves a neutralization step to raise the pH of the crosslinkable mixture to the range of 6 to 8, wherein the neutralization step involves raising the pH of the crosslinkable mixture by no more than 3, no more than 2, or no more than 1.

[0136] Crosslinkable collagen mixtures (comprising collagen compositions, polymers, and additives) can be crosslinked to form collagen gels comprising collagen compositions, polymers, and additives. Crosslinking can be achieved by allowing the crosslinkable collagen mixture to stand for at least 1 second, at least 10 seconds, at least 30 seconds, at least 1 minute, at least 2 minutes, at least 5 minutes, or at least 15 minutes. Typically, the crosslinkable collagen mixture is allowed to stand for at least 30 seconds, at least 1 minute, at least 2 minutes, or at least 5 minutes. Alternatively, the crosslinkable collagen mixture is allowed to stand for at least 30 minutes, at least 1 hour, or at least 2 hours. Alternatively, the crosslinkable collagen mixture is allowed to stand for at least 12 hours, at least 24 hours, 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 temperatures ranging from about 1°C to about 60°C, for example, from about 1°C to about 50°C, from about 1°C to about 30°C, from about 1°C to about 20°C, or from about 1°C to about 10°C. Alternatively, the crosslinkable collagen mixture may be allowed to stand at a temperature of approximately 15°C to approximately 60°C, such as approximately 20°C to approximately 60°C, approximately 20°C to approximately 50°C, or approximately 25°C to approximately 50°C. In one embodiment, the crosslinkable collagen mixture is allowed to stand at a temperature of approximately 4°C. In another embodiment, the crosslinkable collagen mixture is allowed to stand at a temperature of approximately 20°C.

[0137] In some embodiments, the crosslinking step forms chemical crosslinking bonds between the collagen composition and the polymer. In some embodiments, the crosslinking step forms crosslinking bonds within the collagen composition, resulting in a physical network of crosslinked collagen composition entangled or wrapped with the polymer (and additives). In some embodiments, the crosslinking step forms crosslinking bonds between the collagen composition and the polymer, as well as a physically entangled network of crosslinked collagen composition and polymer (and additives).

[0138] Collagen gels are dehydrated to form biomaterials. Suitable dehydrating solvents miscible with water (such as ketones or alcohols) can be used to dehydrate the collagen gel. Typically, collagen gels are dehydrated in acetone or ethanol. Dehydration in a dehydrating solvent such as acetone or ethanol ensures uniform dehydration throughout the collagen gel, particularly preventing one side of the collagen gel from drying faster than the other. Alternatively, the collagen gel can be dehydrated using a dehydrator at temperatures ranging from approximately 25°C to approximately 100°C, for example, from approximately 25°C to approximately 45°C. Typically, the collagen gel is dehydrated using a dehydrator at temperatures ranging from approximately 30°C to approximately 40°C, for example, from approximately 35°C. Alternatively, the collagen gel can be dehydrated using a dehydrator at temperatures ranging from approximately 50°C to approximately 100°C, for example, from approximately 75°C to approximately 100°C, or from approximately 80°C to approximately 100°C. Higher temperature dehydration, such as dehydration at temperatures from about 80°C to about 100°C, is particularly suitable when the crosslinkable collagen mixture contains a foaming agent as an additive; wherein the foaming agent readily decomposes at high temperatures to generate bubbles (e.g., when the foaming agent is sodium bicarbonate). The collagen gel can be dehydrated in a 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% and 35%, for example, between 10% and 35%.

[0139] The shape of the dehydrated collagen gel (i.e., biomaterial) is not limited and can include any two-dimensional or three-dimensional shape. The shape of the biomaterial can be controlled, for example, by crosslinking a crosslinkable collagen mixture in a suitably shaped mold. Alternatively, the collagen gel can be shaped and / or reshaped using suitable forming techniques before and / or after dehydration. Such shaping may 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, dagger-roll coating (KOR coating), slotted die coating, roll-to-roll coating, curtain coating, dip coating, flow coating, spray coating, and brush coating. Coating methods may include coating the collagen gel directly or using an adhesive layer onto a carrier or substrate (such as a paper, polymer, or fabric carrier or substrate), followed by removal of the carrier or substrate after dehydration. Alternatively, the collagen gel can be coated onto a polymer or biopolymer layer that has already been coated onto a carrier or substrate.

[0140] Typically, the biomaterial is formed within a sheet, thus the biomaterial is provided in sheet form. The sheet can be of any thickness, but is typically less than 5 cm thick. Common sheet thicknesses include less than 3 cm, 2 cm, 1 cm, 0.5 cm, 0.2 cm, or 0.1 cm. The sheet can be of uniform thickness or can have varying thicknesses. The sheet can be formed by placing a crosslinkable collagen mixture into a suitable mold to provide the desired thickness and then crosslinking it to form a gel. Alternatively, the sheet can be formed by a continuous manufacturing method, such as coating. The sheet can comprise a single layer of biomaterial or multiple layers of biomaterial, for example, multilayer biomaterials with different properties. The sheet can comprise multiple layers of the same biomaterial, or two or more layers of biomaterial with different compositions.

[0141] The collagen gel formed after cross-linking can be temporarily frozen to facilitate removal from the mold. The gel is then typically thawed before dehydration.

[0142] The biomaterials according to the invention generally have high tensile strength. For example, the biomaterials may have a tensile strength of at least 5 MPa, at least 10 MPa, at least 15 MPa, at least 20 MPa, or at least 25 MPa. Preferably, the biomaterials have a tensile strength of at least 10 MPa, at least 15 MPa, or at least 20 MPa. In one embodiment, the biomaterials have a tensile strength of about 5 MPa to about 25 MPa. Tensile strength is typically measured according to the standard method ISO 3376 (2020).

[0143] Furthermore, the biomaterials according to the invention are generally semi-soft and flexible. The biomaterials typically have a uniform collagen structure throughout their entire thickness. Additionally, the properties of the biomaterials can be easily modified by the amount and type of the polymers, additives, one or more crosslinking agents, and optional other additional components used.

[0144] (c) Processing

[0145] The biomaterial can be further processed to form a processed biomaterial. Therefore, the method described herein may further include a processing step (c) following the sheet-forming step (b) to form the processed biomaterial. The processed biomaterial is typically a leather-like material.

[0146] Processing steps may include any step or combination of steps that produce a leather-like material. As used herein, a leather-like material refers to a material with physical properties similar to natural leather. Typically, a leather-like material is both strong and flexible. When a leather-like material is double-folded, it does not exhibit cracking. Although processing steps are typically performed on biomaterials formed from dehydrated collagen gels, at least fatliquoring and / or dyeing processing steps may be incorporated into the aforementioned gel-forming process. Where fatliquoring and / or dyeing steps are included in the gel-forming process, the biomaterial formed after dehydration of the collagen gel can be a leather-like material that does not require further processing.

[0147] Leather-like materials can have a durability greater than 3 mm, greater than 5 mm, or greater than 7 mm. In one embodiment, the leather-like material has a durability of approximately 5 mm to approximately 10 mm. Durability indicates the distension and strength of the leather grain. Durability is typically measured using a tensile tester according to standard methods ISO 3379 (2015) or DIN 53325.

[0148] Leather-like materials can exhibit lightfastness, meaning that no shading changes or surface degradation are observed after 10, 20, 30, or 40 hours. Lightfastness is typically measured according to the standard method ISO 105-B02 (2014). Lightfastness can also be measured using light with wavelengths of 300-400 nanometers.

[0149] Leather-like materials may possess resistance to environmental aging, such that no shading or surface degradation is observed after 20, 40, or 60 hours of accelerated environmental aging testing. In one embodiment, accelerated environmental aging testing may include placing the biological material at a temperature of 60 ± 2 °C and a humidity of 90 ± 5% RH.

[0150] Leather-like materials may exhibit water stain fastness, meaning that no shading or surface degradation is observed after 10, 13, or 16 hours. Water stain fastness is typically measured according to the standard method ISO 15700 (1998).

[0151] The leather-like material may have a Martindale abrasion resistance of at least 3,000 cycles, at least 4,000 cycles, or at least 5,000 cycles (all measured at 9 kPa). In one embodiment, the leather-like material has a Martindale abrasion resistance of approximately 3,000 to approximately 6,000 cycles at 9 kPa, as measured using a Martindale abrasion tester. Martindale abrasion resistance is typically measured according to the standard method ISO 17076-2 (2011).

[0152] Leather-like materials may have “Veslic” colorfastness to rubbing, meaning that no degradation is observed after 100 wet cycles and 100 dry cycles, 125 wet cycles and 125 dry cycles, or 150 wet cycles and 150 dry cycles. “Veslic” colorfastness to rubbing is usually measured according to the standard method ISO 11640 (2018).

[0153] The leather-like material may have a tensile strength of at least 5 MPa, 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 approximately 5 MPa to approximately 25 MPa. Tensile strength is typically measured according to the standard method ISO 3376 (2020). The elongation of the leather-like material is measured using the same method as for tensile strength, and can be calculated by comparing the length of the leather-like material sample after the tensile strength test with its length before the tensile strength test. The elongation of the leather-like material may be less than 150% of the sample before the tensile strength test, for example, less than 100% or 75% of the sample before the tensile strength test.

[0154] The leather-like material may have a tear strength of at least 5 N, at least 10 N, or at least 15 N. In one embodiment, the leather-like material has a tear strength of approximately 10 N to approximately 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 self-recording tensile testing machine at a lateral kinetic rate of 300 ± 10 mm / min.

[0155] Leather-like materials can possess flexural strength such that no surface degradation is observed after 9,000, 12,000, or 15,000 buckling cycles. Flexural strength is typically measured using a Bally curvature meter according to standard method ISO 5402-1 (2017).

[0156] Leather-like materials may have low levels of heat shrinkage, such as less than 50%, less than 40%, or less than 30%. Preferably, the leather-like material has less than 20% heat shrinkage, more preferably less than 15%. The amount of heat shrinkage is measured by placing the sample at 80°C for 15 minutes. The specimen is typically 20mm x 30mm before the heat shrinkage test. The area of ​​the sample is measured before and after the test, and the shrinkage is calculated as follows:

[0157] Shrinkage (%) = (1 - final area / initial area) * 100

[0158] Leather-like materials may contain low levels of chemical impurities. For example, leather-like materials may contain less than one or more of the following impurities: 75 ppm formaldehyde, 1 ppm chlorophenol, 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 (SVHCs) as defined by the European Chemicals Agency, and 1 ppm organotin compounds.

[0159] This leather-like material can exhibit low swelling levels under high humidity conditions, for example, a swelling ratio (by volume) of less than 90%, less than 80%, or less than 70%. The swelling ratio is tested as follows: the sample is placed at 50°C and 90% relative humidity for 96 hours. The sample volume is measured before and after the test, and the swelling ratio is calculated as follows:

[0160] Swelling (%) = (1 - final volume / initial volume) * 100

[0161] Processing steps may include one or more of fatliquoring, staining, and drying of the biomaterial. Typically, processing step (c) includes fatliquoring, staining, and drying. In one embodiment, the biomaterial is fatliquored, then the fatliquored biomaterial is stained, and then the stained and fatliquored biomaterial is dried. In another embodiment, the biomaterial is fatliquored, and then the fatliquored biomaterial is dried. Processing steps may also include machining the biomaterial. As used herein, machining the biomaterial may include bending, folding, and / or rolling the biomaterial. In one embodiment, the biomaterial is first fatliquored, then the fatliquored biomaterial is stained, then the stained and fatliquored biomaterial is dried, and then the dried biomaterial is processed.

[0162] Lipidification is a process that fixes fats, oils, and / or waxes into the fibers of a material by coating it 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, lipidification may involve contacting a biomaterial or processed biomaterial with an emulsion of vegetable oil in acetone. In the processing of natural leather, lipidification is used to re-oil the leather surface to improve softness and suppleness. Lipidification also adds water-repellent properties. Lipidification of biomaterials formed from marine collagen compositions can produce a hard but brittle material. However, this hard and brittle lipid-added material can still be processed into a leather-like material through further processing steps, particularly through further dyeing, drying, alcohol solution treatment, and / or machining.

[0163] Dyeing can be performed using any suitable dye or pigment. Suitable leather dyes and pigments are known in the art and may include water-based dyes and pigments, alcohol-based dyes and pigments, acid dyes, direct dyes, mordant dyes, or basic 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 conjunction with or instead of a drying step. Typically, the alcohol solution is an ethanol solution.

[0164] The drying step typically involves drying the biomaterial at a temperature of at least 30°C, at least 40°C, or at least 50°C. Drying helps soften the hardened biomaterial resulting from fatliquoring. The drying step can be carried out in a dehydrator. Typically, the water content of the dried biomaterial can be between 5% and 25%.

[0165] Typically, processing also includes finishing and / or coating steps to impart the desired aesthetic appearance to the biomaterial. Suitable finishing chemicals and formulations are known to those skilled in the art and may include waterproofing chemicals, beeswax, or synthetic polymers. If water-based finishing formulations are used, the finished biomaterial must be dried as described above to remove the water solvent.

[0166] Processing may also include any other processes commonly applied to natural leather, including rehydration, splitting, scraping, neutralization, filling, shaping, conditioning, softening, or polishing.

[0167] Manufacturing of products

[0168] Biomaterials or processed biomaterials can be used to manufacture articles comprising biomaterials or processed biomaterials. Manufacturing may include any step of shaping and / or cutting the biomaterials or processed biomaterials. Articles may be made from any available leather-like material, such as accessories, shoes, and furniture.

[0169] Example

[0170] Examples 1 to 10 relate to biomaterials made from collagen compositions. Examples 11, 12, and 14 relate to biomaterials comprising collagen compositions and polymers. Examples 15 to 21 relate to biomaterials according to the present invention comprising collagen compositions, polymers, and additives.

[0171] Example 1

[0172] Leather-like processed biomaterials are prepared according to the methods described below. The biomaterials are made from a collagen composition.

[0173] (a) Extraction

[0174] Cut 80g of cod skin (wet weight) into strips of 3cm x 5cm and rinse with deionized water. Add 250mL of 0.1M NaOH and stir at room temperature for 2 hours (replace the NaOH after 1 hour). Rinse the skin with water.

[0175] Add 200 mL of 10% v / v isopropanol aqueous solution and stir for 1 hour at room temperature. Wash the skin with water.

[0176] Add 200 mL of 1 M AcOH and store overnight at 4°C. Then separate the extract. Add another 200 mL of 1 M AcOH to the fish skin and store overnight again at 4°C.

[0177] The two extracts were combined and freeze-dried to provide a white, amorphous solid. Yield: 100 g / kg (dry weight fish skin).

[0178] (b) Formation

[0179] Dissolve 100 mg of fish skin extract from step (a) in 2 mL of water at room temperature. Add 30 wt% glycerol and 2 wt% glutaraldehyde and mix thoroughly. Transfer the solution to a mold and allow it to stand at 4°C for two days for gel formation.

[0180] Place the gel in the refrigerator for 1 hour and remove it from the mold. After thawing, place the gel in 50 mL of acetone for dehydration (shake at 40 rpm for 48 hours, then replace with fresh acetone after 24 hours).

[0181] After dehydration, the material is semi-soft and flexible.

[0182] (c) Processing

[0183] Fat grafting was performed on the material: The sample was immersed in a 20% v / v vegetable oil solution in acetone and placed on a vibrating table at 40 rpm for 8 hours. The sample was then removed from the solution, and excess oil solution was wiped off. After fat grafting, the material became very hard.

[0184] The fat-added material is then immersed in ethanol-based black leather dye for 1 hour for dyeing. After dyeing, excess dye is wiped off.

[0185] The dyed material was then dried in a dehydrator at 40°C for 5 hours.

[0186] The sample turned into a leather-like texture after being bent multiple times.

[0187] Example 2

[0188] The biomaterial was prepared according to the method of Example 1, but the extraction step (a) was omitted and 100% fish gelatin was used as the starting material for step (b) instead of fish skin extract.

[0189] After completing processing step (c), the resulting material is softer and more elastic than the material produced in Example 1.

[0190] Example 3

[0191] Biomaterials were prepared according to the method of Example 1, but the staining process in processing step (c) was omitted. It can be observed that the hard biomaterials produced after fatliquoring become less hard after drying.

[0192] Example 4

[0193] Biomaterials were prepared according to the method of Example 1, but the staining process in processing step (c) was combined with the formation step (b). In this example, the water-based dye was mixed with the collagen solution before gel formation. It can be observed that the resulting biomaterial is uniformly stained across its thickness.

[0194] Example 5

[0195] The following examples describe methods for creating biomaterials using partially hydrolyzed collagen and optionally fully hydrolyzed collagen without using collagen.

[0196] Partially hydrolyzed collagen and fully hydrolyzed collagen were purchased from Louis Francois and Inner Vita.

[0197] Gel formation

[0198] Dissolve 2.5 g of the protein mixture in 50 mL of degassed type II water at approximately 50-60 °C. After complete dissolution, sonicate the protein mixture for 30 seconds and cool to room temperature (20-25 °C). Add 0.75 g of glycerol (Intralabs) to the protein mixture and stir for 1-2 minutes until completely dissolved. Add 0.2 mL of glutaraldehyde (Alfa Aesar, 2 wt% of protein) to the solution at room temperature and mix for 1-2 minutes before pouring into a 12 cm × 12 cm or 25 cm × 25 cm mold. After 30 minutes, a hydrogel will form at room temperature.

[0199] Dehydration

[0200] The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0201] Example 6

[0202] The following examples describe methods for using partially hydrolyzed collagen and optionally fully hydrolyzed collagen to prepare biomaterials.

[0203] Freeze-dried type I collagen sheets isolated from pig skin and bovine tendons were purchased from Wuxi BIOT Biotechnology Co., Ltd. Marine collagen was extracted from cod skin according to established procedures. Partially hydrolyzed and fully hydrolyzed collagen were purchased from Louis Francois and Inner Vita. XIAMETER AFE-1530 defoamer (silicone-based emulsion) was purchased from Dow Chemical Company.

[0204] Gel formation

[0205] Cut 0.25-0.5g of collagen sheets into small pieces and weigh them for use. Dissolve 2-2.25g of partially hydrolyzed collagen and / or a mixture of partially hydrolyzed and fully hydrolyzed collagen in 25mL of degassed type II water at 50-60°C. Cool the protein solution to room temperature (20-25°C). Add the prepared collagen blocks to 25mL of 0.01M HCl aqueous solution. After the collagen is fully dissolved, add 0.75g of glycerol to the solution. To defoam the solution, add 0.2-0.4% (w / w) of defoaming emulsion and stir the solution further for at least 30 minutes until the foam is completely eliminated. Combine the protein solution and neutralize it by adding an aliquot of 5M NaOH aqueous solution. Add 0.2ml of glutaraldehyde (2w / w%) dropwise and stir the solution for 1-2 minutes before pouring it into a 12cm×12cm or 25cm×25cm mold. After 30 minutes, a hydrogel forms at room temperature.

[0206] Dehydration

[0207] The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0208] Example 7

[0209] Biomaterials containing various collagen components were prepared according to the methods of Examples 5 and 6. The method of Example 5 was used for compositions without collagen, while the method of Example 6 was used for compositions containing collagen. The tensile strength of the biomaterials was tested using an Instron 34SC-05 tensile strength tester according to the standard method ISO 3376:2020.

[0210] Table 1 describes the composition and tensile strength of the biomaterials.

[0211] Table 1: Composition of biomaterials

[0212]

[0213] *A: >20MPa; B: 16-20MPa; C: 11-16MPa; D: 6-11MPa; E: <6MPa

[0214] in conclusion

[0215] The results in Table 1 demonstrate that biomaterials formed from collagen compositions containing partially hydrolyzed collagen (items 3 to 16) exhibit improved tensile strength compared to biomaterials formed from collagen compositions containing collagen as the sole collagen-containing component (items 1 and 2).

[0216] The results also showed that adding defoamer at low concentrations successfully prevented foam formation during the gelation process, thus improving the mechanical strength of the biomaterial.

[0217] Marine collagen is a particularly advantageous source of collagen. For example, biomaterials made from 100% marine collagen (entry 1; 5.4 MPa) exhibit higher tensile strength than biomaterials made from 100% bovine collagen (entry 2; 4.1 MPa).

[0218] Example 8

[0219] Biomaterials containing various collagen components were prepared according to the methods of Examples 5 and 6. A fatliquoring emulsion (40 wt% protein) was added to the collagen composition during gel formation. The results are shown in Table 2.

[0220] Table 2: Tensile strength of biomaterials made from collagen compositions including fat-added emulsions

[0221]

[0222] Example 9

[0223] Biomaterials containing various collagen components were prepared according to the methods of Examples 5 and 6, but the transglutaminase (protein-glutamine γ-glutamyltransferase, EC2.3.2.13) provided by StabizymTGL-100 was used as a cross-linking agent instead of glutaraldehyde.

[0224] Table 3: Composition of biomaterials containing transglutaminase as a cross-linking agent

[0225]

[0226] discuss

[0227] Adding defoamers improves the tensile strength of biomaterials. For example, biomaterials made with the composition of item 2 (0.001 wt% defoamer; 31.5 MPa) exhibited higher tensile strength than those made with item 1 (without defoamer; 25.6 MPa).

[0228] Example 10

[0229] Biomaterials containing various collagen components were prepared according to the method of Example 9. A water-based dye (Metropolitan Leather) was added to the solution during gel formation. The results are shown in Table 4.

[0230] Table 4: Tensile strength of biomaterials made from collagen compositions including water-based dyes

[0231]

[0232] Example 11

[0233] Biomaterials comprising a chemically cross-linked polymer blend of a collagen composition and a polymer are prepared according to the following method.

[0234] Gel formation

[0235] Dissolve a protein mixture (5g, 90% partially hydrolyzed collagen, 10% fully hydrolyzed collagen) and 2%, 5%, 10%, 15%, and 20% (w / w protein) potato starch (Sigma Aldrich) in 50mL of degassed type II water at approximately 50-60°C. After complete dissolution, sonicate the solution for 30 seconds and cool to room temperature (20-25°C). Add 0.75g glycerol (Intralabs) to the protein mixture and stir for 1-2 minutes until completely dissolved. Add 0.38mL glutaraldehyde (Alfa Aesar, 2w / w protein) to the solution at room temperature and mix for 1-2 minutes before pouring into a 12cm x 12cm mold. After 30 minutes, a hydrogel will form at room temperature.

[0236] Dehydration

[0237] The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0238] As shown in Table 5, biomaterials including pectin, alginate, agarose, chitosan, gellan gum and gum arabic were prepared in a similar manner.

[0239] Table 5: Biomaterials comprising polymer blends of collagen compositions and polymers

[0240]

[0241] Example 12

[0242] Biomaterials comprising a physically wound composite material of collagen composition and polymer are prepared according to the following method.

[0243] Gel formation

[0244] A protein mixture (2.5 g, 90% partially hydrolyzed collagen, 10% fully hydrolyzed collagen) was dissolved in 50 mL of degassed type II water at approximately 50-60 °C. After complete dissolution, the solution was sonicated for 30 seconds and cooled to room temperature (20-25 °C). 0.75 g of glycerol (Intralabs) was added to the protein mixture and stirred for 1-2 minutes until completely dissolved. 0.03 g of cotton fibers (pretreated to remove wax) was dispersed in the solution. 25 U (10 U / g) of transglutaminase was added to the solution. After 30 minutes, a hydrogel formed at 50 °C.

[0245] Dehydration

[0246] The collagen hydrogel was dehydrated in a dehydrator at 35°C for 10 hours and then peeled off from the mold.

[0247] As shown in Table 6, biomaterials including nanocellulose fibers, collagen fibers, and keratin fibers were fabricated in a similar manner.

[0248] Table 6: Biomaterials comprising physically wound composites of collagen compositions and polymers

[0249]

[0250] Example 13

[0251] The crosslinkable collagen mixture is placed on a fixed coating paper carrier, which is positioned in front of the blade with a preset gap (0.1 mm to 5 mm, typically 1.2 mm). The blade is then activated, causing the blade to move perpendicular to its edge, thereby uniformly applying the mixture onto the coating paper with a motion parallel to the direction of the coating paper. Once the sample is coated, it is transferred to a dehydration oven, where it is dehydrated at a predetermined temperature for a predetermined time.

[0252] Once dehydration is complete, remove the sample from the coated paper.

[0253] Example 14

[0254] The biomaterials were prepared according to the methods described in Example 9 (“Control”, without polymer) and Example 12 (“Reinforced with cellulose”, where cellulose was used as the polymer). The compositions of the two biomaterials before dehydration are shown below:

[0255] Table 7: Composition of biomaterials

[0256]

[0257] ●The wt.% of the film-forming solution (FFS) is calculated as the weight of the component relative to the weight of the FFS. FFS is defined as partially hydrolyzed collagen + fully hydrolyzed collagen + cellulose, therefore the wt.% of the component relative to the FFS is (component / (partially hydrolyzed collagen + fully hydrolyzed collagen + cellulose) * 100).

[0258] After dehydration, the biomaterial contains approximately 8% water and 10% cellulose.

[0259] Table 8 shows the mechanical properties of the biomaterials.

[0260] Table 8: Mechanical properties of biomaterials.

[0261]

[0262] As shown in Table 8, cellulose-reinforced biomaterials are more flexible than their polymer-free counterparts. Furthermore, cellulose-reinforced biomaterials exhibit lower thermal shrinkage and higher tensile strength than their polymer-free counterparts. Cellulose-reinforced polymers are also softer than polymer-free biomaterials.

[0263] Example 15

[0264] The following examples describe biomaterials prepared using a foaming agent as an additive.

[0265] Biomaterials were prepared using the components described in Table 9. The biomaterials were prepared according to the following method. Unless otherwise stated, all methods began with the dispersion of the protein contents in 100 mL of water. The solution was then heated to 50°C to melt and dissolve the proteins, and this temperature (50°C) was maintained throughout the solution preparation process.

[0266] Entries 9-1 and 9-2 (N2O)

[0267] The solution was stirred throughout the preparation process. After the protein dissolved, the polymer was added at a dry weight of 10% of the protein weight. Plasticizers and fatliquoring agents were then added. If applicable, a humectant was added next. The crosslinking agent was added at a concentration of 10 U / g protein. Immediately after adding the crosslinking agent, the solution was transferred to a sealed container, pressurized with N2O, and agitated to mix. The pressurized mixture was then extruded through a nozzle into a casting mold or onto paper for coating. The foaming solution was then dehydrated to obtain a foamed film.

[0268] Table 9: Composition of biomaterials containing foaming agents

[0269]

[0270] Entries 9-3 to 9-7, 9-12 to 9-16 (Sodium bicarbonate, and optionally SDS)

[0271] Stirring was maintained throughout the solution preparation process. After the protein dissolved, the polymer was added. Sodium dodecyl sulfate (SDS) was then added, if applicable. After the addition of the polymer (and optionally SDS), the plasticizer, humectant, and fatliquoring agent were added in sequence. Subsequently, the dye / pigment was optionally added. Finally, sodium bicarbonate was added. The solution was then treated using one of the following two methods (both methods were used for samples 9-3 to 9-7; the following method (1) was used for samples 9-12 to 9-16):

[0272] 1) Add crosslinking agent. After adding the crosslinking agent, cast or coat the solution into a film. Place the sample at a temperature of 75-95℃ for 20 minutes to allow the sodium bicarbonate to decompose and form bubbles. During this stage, the sample dehydrates and forms foam.

[0273] 2) Add a small amount of 6M HCl (0.2-0.5 mL) to the solution. The acid-base reaction will generate foam, leading to bubble formation. If appropriate, then add a crosslinking agent. After adding the crosslinking agent, cast or coat the solution into a film. Then, dehydrate the sample at 65-85℃.

[0274] Items 9-8, 9-10, 9-11, 9-17 to 9-19 (SDS)

[0275] Keep the solution stirred throughout the preparation process. After the protein dissolves, add the polymer (if appropriate). Add sodium dodecyl sulfate (SDS), followed by the plasticizer and (if appropriate) the humectant and fatliquoring agent. Add the crosslinking agent. Immediately after adding the crosslinking agent, beat the solution using a mechanical whipping device, stirrer, or mixer (such as a benchtop mixer, food processor, or high-shear mixer). After beating but before the crosslinking reaction is complete, cast or coat the solution into a film and then dehydrate it.

[0276] Item 9-9 (Ovalbumin, SDS)

[0277] The method is the same as in entries 9-8, except that the first step involves adding and dissolving the ovalbumin in the solution (before adding the polymer).

[0278] The density of the foamed samples was determined. The density of samples containing N₂O as a foaming agent was measured to be 0.9–1.1 g / cm³. 3 The density of samples containing sodium bicarbonate as a foaming agent ranged from 0.9 to 1.2 g / cm³. 3 This is lower than the density of the corresponding control biomaterial prepared without the addition of a foaming agent, which has a density of 1.2–1.3 g / cm³. 3 .

[0279] Example 16

[0280] The following examples describe biomaterials prepared using powdered fillers as additives.

[0281] The protein contents were dispersed in 50 mL of water at a concentration of 10% protein (w / v). The solution was then heated to 50°C to dissolve the protein, and this temperature (50°C) was maintained throughout the solution preparation process. Cellulose fibers and plasticizers were added under continuous mixing at the concentrations shown in Table 10. Powdered filler was added and mixed with a high-shear mixer for at least 1 minute. Crosslinking agent was added under continuous mixing. Immediately after the addition of crosslinking agent, the solution was cast or coated into a film and subsequently dehydrated.

[0282] Table 10: Composition of biomaterials containing powdered fillers

[0283]

[0284] Example 17

[0285] The following examples illustrate the performance of biomaterials containing powdered fillers as additives.

[0286] Biomaterials were prepared according to the method of Example 16, and their composition is shown in Table 11. The powdered fillers used were cork powder, colloidal silica, zeolite, calcium carbonate, and titanium dioxide.

[0287] Table 11: Composition of biomaterials containing powdered fillers used for mechanical testing

[0288]

[0289] The mechanical test results are shown in Table 12.

[0290] Table 12: Mechanical testing of biomaterials containing powdered fillers

[0291]

[0292] The results showed that, compared with biomaterials without powdered fillers, biomaterials with powdered fillers exhibited reduced elongation and improved thermal stability.

[0293] Example 18

[0294] The following examples describe biomaterials prepared using anti-hydrolysis agents as additives.

[0295] The protein contents were dispersed in 50 mL of water at a concentration of 10% protein (w / v). The solution was then heated to 50°C to dissolve the protein, and this temperature (50°C) was maintained throughout the solution preparation process. Cellulose fibers and plasticizers were added under continuous mixing at the concentrations shown in Table 13. An anti-hydrolysis agent was added, and the mixture was stirred at 800 rpm for at least 3 minutes using a high-speed mixer. A cross-linking agent was added under continuous mixing. Immediately after adding the cross-linking agent, the solution was cast or coated into a film, followed by dehydration.

[0296] Table 13: Composition of biomaterials containing anti-hydrolysis agents

[0297]

[0298] Example 19

[0299] The following examples describe the performance of biomaterials containing anti-hydrolysis agents as additives.

[0300] Biomaterials were prepared according to the method of Example 18, and their composition is shown in Table 14. The anti-hydrolysis agent used was Permutex. XR-13-906 (polycarbodiimide), V1094, V1074, and LUBIO AS 15 (aliphatic carbodiimide).

[0301] Table 14: Composition of biomaterials containing anti-hydrolysis agents for mechanical testing

[0302]

[0303] The mechanical test results are shown in Table 15.

[0304] The results showed that, compared with biomaterials without hydrolysis inhibitors, biomaterials with hydrolysis inhibitors exhibited improved thermal stability and reduced swelling in high humidity environments.

[0305] Table 15: Mechanical testing of biomaterials containing anti-hydrolysis agents

[0306]

[0307] Example 20

[0308] The following examples describe biomaterials prepared using low-density fillers as additives.

[0309] The protein contents were dispersed in 100 mL of water at a concentration of 25% protein (wt. / wt.). The solution was then heated to 50°C to melt and dissolve the protein, and this temperature (50°C) was maintained throughout the solution preparation process. Stirring was maintained throughout the solution preparation. After the protein dissolved, cellulose was added at the concentrations shown in Table 16. Following the addition of cellulose, plasticizers, humectants, fatliquoring agents, and dyes were added sequentially. Low-density fillers were then added. An antifoaming agent was then added to the solution, and the solution was degassed under vacuum stirring. Finally, a crosslinking agent was added to the solution, which was then coated and dehydrated at 65-85°C.

[0310] Table 16: Composition of biomaterials containing low-density fillers

[0311]

[0312] Example 21

[0313] The following examples describe the performance of biomaterials containing low-density fillers as additives.

[0314] Biomaterials were prepared according to the method of Example 20, and their composition is shown in Table 17. The low-density fillers used were glass bubbles (60 µm and 30 µm in diameter).

[0315] Table 17: Composition of biomaterials containing low-density fillers used for mechanical testing

[0316]

[0317] The results of the mechanical tests are shown in Table 18.

[0318] The results showed that biomaterials containing low-density fillers exhibited reduced elongation compared to biomaterials without low-density fillers.

[0319] Table 18: Mechanical Testing of Biomaterials Containing Low-Density Fillers

[0320]

Claims

1. A biomaterial comprising a dehydrated collagen gel, wherein the collagen gel comprises (a) a collagen composition, (b) a polymer and (c) an additive; The collagen composition comprises (i) partially hydrolyzed collagen and (ii) collagen and / or fully hydrolyzed collagen. The collagen composition comprises at least 30% by weight of partially hydrolyzed collagen, and The additives mentioned therein are selected from anti-hydrolysis agents, foaming agents, thickeners, fillers and flame retardants.

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

4. The biomaterial according to any one of the preceding claims, wherein the collagen composition comprises 70% to 95% by weight of partially hydrolyzed collagen.

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

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

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

8. The biomaterial according to any one of claims 1 to 4, wherein the collagen composition comprises 5% to 30% by weight of fully hydrolyzed collagen, preferably, wherein the collagen composition comprises 5% to 20% by weight of fully hydrolyzed collagen.

9. The biomaterial according to any one of the preceding claims, wherein the collagen gel comprises a polymer blend, the polymer blend comprising the collagen composition and the polymer.

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

11. The biomaterial according to any one of claims 1 to 8, wherein the collagen gel comprises a composite material comprising the collagen composition and the polymer.

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

13. The biomaterial according to claim 12, wherein the cellulose fiber is a micron-sized cellulose fiber or a nano-sized cellulose fiber.

14. The biomaterial according to any one of the preceding claims, wherein the collagen composition further comprises an antifoaming agent.

15. The biomaterial of claim 14, wherein the defoamer is present in an amount from 0.001 wt% to 5 wt%.

16. The biomaterial of claim 15, wherein the defoamer is present in an amount of 0.1 wt% to 2 wt%.

17. The biomaterial according to any one of the preceding claims, wherein the additive is an anti-hydrolysis agent.

18. The biomaterial according to any one of claims 1 to 17, wherein the anti-hydrolysis agent is selected from polycarbodiimide and aliphatic carbodiimide.

19. The biomaterial according to any one of claims 1 to 16, wherein the additive is a foaming agent.

20. The biomaterial according to any one of claims 1 to 16 or claim 19, wherein the foaming agent is selected from NO2, sodium bicarbonate, sodium dodecyl sulfate and ovalbumin.

21. The biomaterial according to any one of claims 1 to 16, wherein the additive is a filler.

22. The biomaterial according to any one of claims 1 to 16 or claim 21, wherein the filler is a powdered filler or a low-density filler.

23. The biomaterial according to any one of claims 1 to 16, 21 or 22, wherein the filler is a powdered filler selected from cork powder, colloidal silica, zeolite, calcium carbonate and titanium dioxide.

24. The biomaterial according to any one of claims 1 to 16, 21 or 22, wherein the filler is a low-density filler selected from glass bubbles and fumed silica.

25. The biomaterial according to any one of the preceding claims, wherein the collagen gel comprises two or more additives, each additive being independently selected from anti-hydrolysis agents, foaming agents, thickeners, fillers, and flame retardants.

26. The biomaterial according to any of the preceding claims, wherein at least one of the partially hydrolyzed collagen, collagen and fully hydrolyzed collagen is extracted from seafood.

27. A method for producing biomaterials according to any one of claims 1 to 26, the method comprising: a) Forming a collagen gel comprising a collagen composition, polymers, and additives; and b) dehydrating the collagen gel to form the biomaterial.

28. The method of claim 27, wherein the forming step comprises adding a fatliquoring component and / or a dye or pigment to the collagen composition and / or polymer.

29. The method of claim 27 or claim 28, wherein the method further comprises processing the biomaterial to form processed biomaterial.

30. The method according to any one of claims 27 to 29, further comprising adding a textile liner to the biomaterial or processed biomaterial.

31. A leather-like processed biomaterial comprising the biomaterial according to any one of claims 1 to 26.

32. A composite material comprising the biomaterial according to any one of claims 1 to 26 or the leather-like processed biomaterial according to claim 31, and a textile lining.

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