Helically organized silk fibroin fiber bundles for matrices in tissue engineering

• US 6,902,932 B2
• Filed: 11/16/2001
• Issued: 06/07/2005
• Est. Priority Date: 11/16/2001
• Status: Expired due to Term

• Current Assignee: Allergan, Inc. (Canada) (Abbvie Incorporated), Trustees of Tufts College
• Original Assignee: Tissue Regeneration, Inc. (Abbvie Incorporated), Trustees of Tufts College
• Inventors: Horan, Rebecca L., Altman, Gregory H., Horan, David J., Kaplan, David L.
• Primary Examiner(s): Naff, David M.
• Application Number:
  US10/008,924
• Publication Number:
  US 20030100108A1
• Time in Patent Office: 1,299 Days
• Field of Search: 435/174, 435/177, 435/180, 435/182, 435/395, 424/423, 424/93.7
• US Class Current: 435/395
• 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 silk-fiber-based matrix composition comprising sericin-extracted silkworm fibroin fibers, said fibers being biocompatible and helically organized into fiber bundles, wherein said matrix supports ingrowth of cells around said fibroin fibers and is biodegradable.

2. The matrix as recited in claim 1, wherein the silk-fiber based matrix comprises fibroin fibers obtained from Bombyx mori silkworm.

3. The matrix as recited in claim 1, wherein the matrix comprises a composite of the sericin-extracted fibroin fibers and collagen fibers.

4. The matrix as recited in claim 1, wherein the matrix comprises a composite of the sericin-extracted fibroin fibers and one or more silk foams, films, meshes or sponges.

5. The matrix as recited in claim 1, wherein the matrix comprises a composite of the sericin-extracted fibroin fibers and one or more degradable polymers selected from group consisting of Collagens, Polylactic acid or its copolymers, Polyglycolic acid or its copolymers, Polyanhydrides, Elastin, Glycosamino glycans, and Polysaccharides.

6. The matrix as recited in claim 1, further comprising pluripotent or fibroblast cells seeded on said matrix.

7. The matrix as recited in claim 6, wherein said pluripotent or fibroblast cells are autologous.

8. The matrix as recited in claim 7, wherein said pluripotent or fibroblast cells are allogeneic.

9. The matrix as recited in claim 7, wherein said pluripotent cells are selected from the group consisting of bone marrow stromal cells and adult or embryonic stem cells.

10. The matrix as recited in claim 7, wherein said fibroblast cells are mature human ACL fibroblast cells.

11. The matrix as recited in claim 7, wherein said pluripotent or fibroblast cells proliferate and differentiate on said matrix to form said predetermined ligament or tendon.

12. The matrix as recited in claim 7, further comprising a surface modification agent which enhances proliferation and differentiation of said pluripotent or fibroblast cells on said matrix.

13. The matrix as recited in claim 1, wherein said matrix comprises a shape of a ligament or tendon selected from the group consisting of anterior cruciate ligament, posterior cruciate ligament, rotator cuff tendon, medial collateral ligament of the elbow, flexor tendon of the hand, ligaments and tendons of the temporomandibular joint, and lateral ligament of the ankle.

14. The matrix as recited in claim 13, wherein said ligament is an anterior cruciate ligament.

15. The matrix of claim 1, wherein said helically-organized fibers comprise a cylindrical shape.

16. The matrix of claim 12, wherein said surface modification comprises an arginine-glycine-aspartate peptide.

17. The matrix of claim 1, wherein said matrix comprises an ultimate tensile strength of greater than 2000N and a linear stiffness of between 100-600N/mm.

18. A silk-fiber-based matrix comprising helically organized fiber bundles of sericin-extracted biodegradable silkworm fibroin fibers and bone marrow stromal cells attached thereto.

19. A composition consisting essentially of sericin-extracted silkworm fibroin fibers, said fibers being biodegradable and organized into a matrix being helically organized into fiber bundles and comprising an ultimate tensile strength of greater than 2000N and a linear stiffness of between 100-600N/mm, wherein sericin is completely removed from said fiber.