Silk fibroin fiber bundles for matrices in tissue engineering

• US 20050089552A1
• Filed: 11/16/2004
• Published: 04/28/2005
• Est. Priority Date: 11/16/2001
• Status: Abandoned Application

• Current Assignee: Allergan Incorporated (Abbvie Incorporated)
• Original Assignee: Tissue Regeneration, Inc. (Abbvie Incorporated), Trustees of Tufts College
• Inventors: Horan, Rebecca L., Altman, Gregory H., Horan, David J., Kaplan, David L.
• Application Number:
  US10/990,199
• Publication Number:
  US 20050089552A1
• Time in Patent Office: Days
• US Class Current: 424/445
• CPC Class Codes: A61F 2/00 Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents as cosmetic articles, see the relevant subclasses, e.g. wigs, hair pieces, A41G3/00, A41G5/00; artificial nails A45D31/00; dental prostheses A61C13/00; materials for prostheses A61L27/00; artificial hearts A61M60/00; artificial kidneys A61M1/14
• CPC Class Codes: A61F 2/08 Muscles; Tendons; Ligaments sutures A61B17/04
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 9/70 Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug hollow drug-filled fibres A61K9/0092
• CPC Class Codes: A61L 2430/10 for reconstruction of tendons or ligaments
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/3608 Bone, e.g. demineralised bone matrix [DBM], bone powder
• CPC Class Codes: A61L 27/3662 Ligaments, tendons
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3821 Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61L 27/386 Ligaments, tendons
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/48 with macromolecular fillers
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 14/43518 : from spiders
• CPC Class Codes: C08L 89/00 : Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00
• CPC Class Codes: C12N 2533/50 : Proteins
• CPC Class Codes: C12N 5/066 : Tenocytes; Tendons, Ligaments
• CPC Class Codes: C12N 5/0663 : Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: Y10T 428/249921 : Web or sheet containing structurally defined element or component
• CPC Class Codes: Y10T 428/298 : Physical dimension
• CPC Class Codes: Y10T 442/2566 : Organic solvent resistant [e.g., dry cleaning fluid, etc.]

Abstract

The present invention provides a novel silk-fiber-based matrix having a wire-rope geometry for use in producing a ligament or tendon, particularly an anterior cruciate ligament, ex vivo for implantation into a recipient in need thereof. The invention further provides the novel silk-fiber-based matrix which is seeded with pluripotent cells that proliferate and differentiate on the matrix to form a ligament or tendon ex vivo. Also disclosed is a bioengineered ligament comprising the silk-fiber-based matrix seeded with pluripotent cells that proliferate and differentiate on the matrix to form the ligament or tendon. A method for producing a ligament or tendon ex vivo comprising the novel silk-fiber-based matrix is also disclosed.

Claims

1. A fiber construct comprising sericin-extracted silkworm fibroin fibers, wherein the fibers have a diameter of about 20 to about 40 μ

m and an average ultimate tensile strength of at least about 0.67 N/fiber.

2. The fiber construct of claim 1, wherein the fibers are organized in parallel.

3. The fiber construct of claim 1, wherein the fibers are organized in a helical, wire rope, twisted, braided, mesh-like or cabled geometry.

4. The fiber construct of claim 1, wherein the fibers comprise a coating and/or surface modification that promotes cellular attachment and/or tissue differentiation and proliferation thereon.

5. The fiber construct of claim 4, wherein said coating and/or surface modification comprises an arginine-glycine-aspartate (RGD) peptide.

6. The fiber construct of claim 4, wherein said coating and/or surface modification comprises a growth factor.

7. The fiber construct of claim 1, wherein the construct comprises a group of at least one to about 1300 fibroin fibers.

8. The fiber construct of claim 7, wherein the construct further comprises at least two groups forming a bundle.

9. The fiber construct of claim 8, wherein the construct further comprises at least two bundles forming a strand.

10. The fiber construct of claim 9, wherein the construct further comprises at least two strands forming a cord.

11. The fiber construct of claim 7, wherein the group comprises fibroin fibers organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry.

12. The fiber construct of claim 8, wherein the bundle comprises groups organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry.

13. The fiber construct of claim 9, wherein the strand comprises bundles organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry.

14. The fiber construct of claim 10, wherein the cord comprises strands organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry.

15. The fiber construct of claim 1, wherein the construct has an average ultimate tensile strength of at least about 6.67 N.

16. The fiber construct of claim 1, further comprising cells.

17. The fiber construct of claim 16, wherein the cells are selected from the group consisting of stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells.

18. A method for producing a sericin-extracted silk-fibroin fiber construct comprising:

contacting at least one silkworm fibroin fiber with an aqueous solution of Na₂Co₃ and detergent to extract sericin from the fiber.

23. The method as in one of claims 18-22, wherein the sericin is extracted at a temperature no greater than about 90°

C.

24. The method as in one of claims 18-22, further comprising the step of coating the fibroin fibers with a coating and/or surface modifier that promotes cellular attachment and/or tissue proliferation on the fibers.

25. The method of claim 24, wherein said coating and/or surface modifier comprises an arginine-glycine-aspartate (RGD) peptide.

26. The method of claim 24, wherein said coating and/or surface modifier comprises a growth factor.

28. The method of claim 18, further comprising the step of placing the fiber in a twisted, helical, braided, mesh-like or cabled geometry.

33. The method as in any of claims 18-22, further comprising:

a. contacting the construct with cells; and

b. culturing the construct under conditions suitable for cell growth and regeneration.

34. The method of claim 33, wherein the cells are stem cells, muscle cells, bone marrow stromal cells, pluripotent cells, or fibroblast cells.

19. A method for producing a sericin-extracted silk-fibroin fiber construct comprising:

a. arranging at least 2 silkworm fibroin fibers to form a group; and

b. contacting the group with an aqueous solution of Na₂Co₃ and detergent to extract sericin from the fibers.

27. The method of claim 19, wherein said group comprises up to 1300 fibroin fibers.

29. The method of claim 19, wherein the fibers are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry.

20. A method for producing a sericin-extracted silk-fibroin fiber construct comprising:

a. arranging at least 2 groups of silkworm fibroin fibers to form a bundle; and

b. contacting the bundle with an aqueous solution of Na2Co3 and detergent to extract sericin from the fibers.

30. The method of claim 20, wherein the groups are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a bundle.

21. A method for producing a sericin-extracted silk-fibroin fiber construct comprising:

a. arranging at least 2 bundles of silkworm fibroin fibers to form a strand; and

b. contacting the strand with an aqueous solution of Na2Co3 and detergent to extract sericin from the fibers.

31. The method of claim 21, wherein the bundles are organized in a parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a strand.

22. A method for producing a sericin-extracted silk-fibroin fiber construct comprising:

a. arranging at least 2 strands of silkworm fibroin fibers to form a cord; and

b. contacting the cord with an aqueous solution of Na2Co3 and detergent to extract sericin from the fibers.

32. The method of claim 22, wherein the strands are organized in a in parallel, helical, wire rope, twisted, braided, mesh-like or cabled geometry forming a cord.