Method for making a biofabricated material containing collagen fibrils
Described herein is a method for producing a biofabricated material from collagen or collagen-like proteins which are recombinantly produced and which contain substantially no 3-hydroxyproline. The collagen or collagen-like proteins are isolated from animal sources, or produced by recombinant DNA techniques or by chemical synthesis. The collagen or collagen-like proteins are fibrillated, crosslinked, dehydrated and lubricated thus forming the biofabricated material having a substantially uniform network of collagen fibrils.
1. A method for making a material comprising a network of recombinant non-human collagen fibrils, wherein said material contains no more than 25% by weight water; said material has a collagen density of at least 70 mg/cc; and wherein said material contains at least 1% of a lubricant, comprising:
fibrillating an aqueous solution or suspension of isolated or purified recombinant non-human collagen molecules into collagen fibrils,
crosslinking said collagen fibrils by contacting them with at least one crosslinking agent, and
dehydrating the crosslinked collagen fibrils so that they contain less than 25% by weight water, and
further incorporating at least 1% by weight of at least one lubricant prior to, during, or after any of said fibrillating, crosslinking, and dehydrating.
2. The method of claim 1 , further comprising staking the material.
3. The method of claim 1 , further comprising casting, molding or forming the material
into one having a top and bottom surface or an inner and outer surface.
4. The method of claim 1 , wherein the non-human collagen molecules are recombinantly expressed in Escherichia coli.
5. The method of claim 1 , wherein the non-human collagen molecules are recombinantly expressed in Pichia.
6. The method of claim 1 , wherein the collagen comprises Type I collagen.
7. The method of claim 1 , wherein the collagen comprises Type III collagen.
8. The method of claim 1 , wherein fibrillating comprises adding a salt or a combination of salts.
9. The method of claim 1 , wherein fibrillating comprises adjusting the pH to pH 5.5 or more with an acid, a base, or a buffer.
10. The method of claim 1 , wherein said crosslinker is at least one selected from the group consisting of an amine, carboxylic acid, sulfate, sulfite, sulfonate, aldehyde, hydrazide, sulfhydryl, diazirine, aryl, azide, acrylate, epoxide, phenol, chromium compound, vegetable tannin, and syntan.
11. The method of claim 1 that comprises dehydrating the network of collagen fibrils by contacting them with an agent that removes bound water from collagen.
12. The method of claim 1 that comprises dehydrating the network of collagen fibrils by contacting them with at least one dehydrating or dewatering solvent.
13. The method of claim 1 that comprises dehydrating the network of collagen fibrils by contacting them with at least one dehydrating or dewatering solvent comprising acetone.
14. The method according to claim 1 , further comprising uniformly applying the at least one lubricant on or throughout the material, wherein the concentration by weight of the at least one lubricant on or in identical unit volumes of the material varies by no more than 20%.
15. The method according to claim 1 , wherein the lubricant is selected from the group consisting of at least one fat, biological, mineral or synthetic oil, cod oil, sulfonated oil, polymer, and organofunctional siloxane.
16. The method according to claim 1 , further comprising uniformly distributing a dye, stain, pigment, resin, polymer, or paint on or throughout the material, wherein the concentration by weight of dye, stain, pigment, resin, polymer, or paint in identical unit volumes of the material varies by no more than 20%.
17. The method according to claim 1 , further comprising uniformly applying a surface coating or surface finish on or throughout the material, wherein the concentration by weight of the surface coating or surface finish on or in identical unit volumes of the material varies by no more than 20%.
18. The method according to claim 1 , wherein the crosslinked and lubricated collagen fibrils are distributed uniformly throughout the material, wherein the weight of the fibrils in identical unit volumes of the material varies by no more than 20%.
19. The method according to claim 1 that further comprises incorporating at least one filler into said material; wherein the concentration of the filler by weight in or on identical unit volumes of the material varies by no more than 20%.
20. The method according to claim 1 , further comprising incorporating into or onto said material at least one woven or nonwoven material or further comprising incorporating said material into at least one woven or nonwoven material.
21. The method according to claim 1 , wherein the collagen contains substantially no 3-hydroxyproline.
22. The method according to claim 1 , wherein the collagen contains substantially no hydroxylysine.
23. The method according to claim 1 , wherein the network of recombinant non-human collagen fibrils have no more than 40% by weight of the collagen fibrils in the form of collagen fibers having a diameter of 10 μm or more.
24. The method according to claim 1 , wherein the network of recombinant non-human collagen fibrils have no more than 10% by weight of the collagen fibrils in the form of collagen fibers having a diameter of 5 μm or more.
25. The method according to claim 1 , wherein said material comprises no more than 1% of actin.
26. The method according to claim 1 , wherein said method is performed for a time and under conditions that produce a material having a tensile strength of at least 1 MPa, a flexibility determined by elongation at break of at least 1%, a softness as determined by ISO 17235 of at least 2 mm, a thickness of at least 0.05 mm.
27. The method of claim 1 , wherein said recombinant non-human collagen molecules are isolated from cells that are cultured in vitro.
28. The method of claim 1 , wherein said recombinant non-human collagen molecules are purified.
29. The method of claim 28 , wherein said recombinant non-human collagen molecules have a degree of purity of at least 40% by weight based on its total protein content.
30. The method according to claim 1 , wherein said material contains at least 10% by weight water.
31. The method according to claim 1 , wherein said material has a collagen density of at least 100 mg/cc.
32. The method according to claim 1 , wherein said material has a collagen density of 100 mg/cc to 500 mg/cc.
33. A method for making a material comprising a network of recombinant collagen fibrils, wherein said material contains no more than 25% by weight water and no more than 1% of actin; said material has a collagen density of at least 70 mg/cc; and wherein said material contains at least 1% of a lubricant, comprising:
fibrillating an aqueous solution or suspension of isolated or purified recombinant collagen molecules into collagen fibrils,
crosslinking said collagen fibrils by contacting them with at least one crosslinking agent, and
dehydrating the crosslinked collagen fibrils so that they contain less than 25% by weight water, and
further incorporating at least 1% by weight of at least one lubricant prior to, during, or after any of said fibrillating, crosslinking, and dehydrating.
34. The method according to claim 33 , wherein said material contains at least 10% by weight water.
35. The method according to claim 33 , wherein said material has a collagen density of at least 100 mg/cc.
36. The method according to claim 33 , wherein said material has a collagen density of 100 mg/cc to 500 mg/cc.