Scaffold formed of tissue treated to eliminate cellular and cytosolic elements

• US 6,652,872 B2
• Filed: 04/04/2001
• Issued: 11/25/2003
• Est. Priority Date: 07/06/1999
• Status: Expired due to Fees

• Current Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Original Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Inventors: Nevo, Zvi, Robinson, Dror
• Primary Examiner(s): Naff, David M.
• Application Number:
  US09/826,389
• Publication Number:
  US 20020009805A1
• Time in Patent Office: 965 Days
• Field of Search: 435/174, 435/177, 435/395, 424/423, 424/93.7, 623/11.11
• US Class Current: 424/423
• CPC Class Codes: A61F 2/3094 Designing or manufacturing processes
• CPC Class Codes: A61F 2002/30762 Means for culturing cartilage
• CPC Class Codes: A61F 2002/30766 Scaffolds for cartilage ingrowth and regeneration
• 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: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3608 Bone, e.g. demineralised bone matrix [DBM], bone powder
• CPC Class Codes: A61L 27/3612 Cartilage, synovial fluid
• CPC Class Codes: A61L 27/3629 Intestinal tissue, e.g. small intestinal submucosa
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 2533/92 Amnion; Decellularised dermis or mucosa
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

Abstract

A scaffold for use as a growth supportive base for cells and tissue explants is formed of naturally occurring derived connective or skeletal tissue that has been treated for elimination of cellular and cytosolic components comprising DNA, RNA, peptides, lipids and proteoglycans. The treated tissue can be crosslinked and treated for producing a higher porosity such as by mechanical or chemical treatment. Alternatively, the treated tissue is ground to form flakes having a size below 1000μ, and the flakes are attached to each other to provide a scaffold having a porosity of at least 85%. The scaffold may be composed of several single-flake layers arranged on top of the other. The scaffold can be impregnated with cells to form tissue such as epiphysial tissue, and may contain adhesive molecules or growth factors. A prosthesis for implanting can be formed using the scaffold. The scaffold may be composed of fused epiphyses.

Claims

1. A scaffold for use as growth supportive base for cells and tissue explants from three-dimensional tissue, comprising:

a naturally occurring derived connective or skeletal tissue which has been treated for elimination of cellular and cytosolic elements comprising DNA, RNA, peptides, lipids and proteoglycans, and which has been grinded to form flakes having a size below 1000μ

, which flakes are attached to each other, the scaffold having a porosity of at least 85%.

2. A scaffold according to claim 1, having at least 90% porosity.

3. A scaffold according to claim 2, having at least 95%-98% porosity.

4. A scaffold according to claim 1, where the naturally occurring connective or skeletal tissue is selected from the group consisting of:

cartilage, epiphysis, perichondrium, periosteum, meniscus, ligaments, skin, demineralized bone and subinternal layer of intestine.

5. A scaffold according to claim 1, further comprising at least one compound selected from the group consisting of:

adhesive molecules and growth factors.

6. A scaffold according to claim 5, wherein the adhesive molecules are selected from the group consisting of:

integrines, proteoglycan, glycosaminoglycans, laminin, fibronectin, hyaluronic acid, polylysine and lysozyme.

7. A scaffold according to claim 5, wherein the growth factors are selected from the group consisting of:

FGF'"'"'s, TGF'"'"'s, BMP'"'"'s, and IGF'"'"'s.

8. A scaffold according to claim 1, impregnated with cells forming a three-dimensional tissue.

9. A scaffold according to claim 8, wherein the three-dimensional tissue is composed of cells from autologous or allogeneic sources selected from the group consisting of:

mesenchymal tissue, articular cartilage, epiphyseal tissue, perichondrial and periosteal flaps, skeletal tissue, liver and kidney tissues, neuronal tissue, dermis tissue and fibrous tissue, cells from an early embryonal stage, and stem progenitor cells from young and adult tissues.

10. A scaffold according to claim 1, wherein said three-dimensional tissue comprises epiphysial tissue.

11. A scaffold according to claim 10, wherein said epiphysial tissue is obtained from 17-22 week old fetus.

12. A prosthesis for implanting into a body of an individual comprising as an active ingredient the scaffold of claim 8.

13. A prosthesis for implanting into a body of an individual comprising as an active ingredient the scaffold of claim 9.

14. A prothesis according to claim 12, formed by at least 5 single-flake layers of the scaffold stacked on each other.

15. A prosthesis according to claim 14, wherein the edges of the layers are fused to each other, by thermo treatment.

16. A prosthesis according to claim 14, wherein the edges of the layers are fused to each other, by addition of adhesive molecules.