REPAIR OF CARTILAGE TISSUE USING A MATRIX GEL CONTAINING CHONDROCYTES

• US 20100047316A1
• Filed: 08/31/2007
• Published: 02/25/2010
• Est. Priority Date: 08/31/2006
• Status: Active Grant

• Current Assignee: CartiRegen B.V.
• Original Assignee: Cellcotec B.V. (CellCoTec Ltd.)
• Inventors: Riesle, Jens Uwe, Hendriks, Jeanine Anna Alphonse
• Granted Patent: US 9,072,819 B2
• Time in Patent Office: Days
• US Class Current: 424/425
• CPC Class Codes: A61L 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3817 Cartilage-forming cells, e.g. pre-chondrocytes
• CPC Class Codes: A61L 27/3852 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: C12N 2501/00 Active agents used in cell culture processes, e.g. differentation
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

Abstract

The present invention relates in one aspect to the use of a matrix gel comprising chondrocytes or progenitor cells thereof in a density below that of natural cartilage as a cartilage repair implant wherein said cells exhibit increases production of extracellular matrix material.

Claims

1. A method to repair a cartilage defect in a mammal which method comprises implanting into a cartilage defect in said subject a matrix gel comprising chondrocytes said chondrocytes isolated from a cartilage sample, concentrated and mixed with a matrix gel material wherein said chondrocytes in said matrix gel exhibit increased production of extracellular matrix material;

or implanting into said defect said matrix loaded on a scaffold.

2. The method of claim 1, wherein said matrix gel comprising chondrocytes is essentially free of chondrons.

3. The method of claim 1, wherein said extracellular matrix material is GAG.

4. The method of claim 1, to induce production of cartilage extracellular matrix and/or cartilage tissue.

5. The method of claim 1, wherein said matrix gel comprises a fibrin glue, hyaluronic acid, alginate, propylene fumarate-co-ethylene glycol hydrogel, hydrogypropyl methylcellulose hydrogel, collagen mimetic peptide, chitosan, collagen, gelatin, (ethylene glycol)-co-poly(lactic acid)hydrogel, agarose, Pluronic™

F-127, copolymers of poly(ethylene glycol)-terephthalate (PEGT) and poly(butylene terephthalate) (PBT), or combinations thereof.

6. The method of claim 1, wherein the chondrocyte density in said matrix gel is 0.8 to 40,000 times lower, preferably 8 to 5,000 times lower than that of the isolate as a result of said dilution.

7. The method of claim 1, wherein said chondrocytes are autologous chondrocytes obtained from an autologous cartilage sample.

8. The method of claim 1, wherein said chondrocytes are obtained by separating said chondrocytes from said cartilage sample by electrophoretic separation, or by digestive treatment.

9. The method of claim 8, wherein said digestion is performed in a digestion solution comprising at least one enzyme selected from the group consisting of collagenase, dispase, trypsin, hyaluronidase, chondroitinase ABC, elastase, heparitinase and alpha-chymotrypsin.

10. The method of claim 9 wherein said enzyme is collagenase type II.

11. The method of claim 1, wherein said cartilage sample comprises articular cartilage.

13. The method of claim 1, wherein said mammal has a cartilage disorder which is articular cartilage trauma, meniscus injury, chondrogenesis disorders or arthritis.

12. (canceled)

14. Method for preparing an implant for repairing a cartilage defect in a mammal in need thereof comprising the steps of:

a) providing a sample comprising chondrocytes;

b) isolating said chondrocytes from said sample;

c) concentrating the chondrocytes;

d) mixing the isolated chondrocytes with a matrix gel material to obtain a diluted isolate wherein said cells exhibit increases production of extracellular matrix material; and

e) optionally loading said matrix gel comprising said chondrocytes in or on a scaffold.

15. The method of claim 14, wherein said sample comprises autologous chondrocytes obtained from articular cartilage, preferably from a body part comprising the site of the defect.

16. The method of claim 14, wherein said step b) comprises isolating said chondrocytes from said sample by electrophoretic separation.

17. The method of claim 16, wherein said electrophoretic separation is performed by

a) mincing said sample to fragments;

b) optionally subjecting said fragments to enzymatic digestion;

c) submerging said optionally digested fragments in an electrophoresis gel;

d) subjecting said submerged fragments to electrophoresis conditions, and

e) isolating said chondrocytes from the electrophoresis buffer.

18. The method of claim 14, wherein said matrix gel material is a biocompatible matrix gel material, preferably selected from the group consisting of fibrin glue, hyaluronic acid, alginate, propylene fumarate-co-ethylene glycol hydrogel, hydrogypropyl methylcellulose hydrogel, collagen mimetic peptide, chitosan, collagen, gelatin, (ethylene glycol)-co-poly(lactic acid)hydrogel, agarose, Pluronic™

F-127, copolymers of poly(ethylene glycol)-terephthalate (PEGT) and poly(butylene terephthalate) (PBT), and combinations thereof.

19. The method of claim 14, wherein said cells are mixed with said matrix gel material to result in a cell density corresponding to a dilution of 0.8 to 40,000 times, preferably 8 to 5,000 times relative to that of the autologous source.

20. The method of claim 14, wherein said increased production of extracellular matrix material is exhibited by an increase in the GAG/DNA ratio within said matrix gel.

21. A matrix gel comprising autologous chondrocytes at a density below that at which the chondrocytes are present in the cartilage from which the chondrocytes were obtained produced by the method according to claim 14.

22. A scaffold comprising the matrix gel of claim 21.

23. A matrix gel comprising chondrocytes isolated from a cartilage sample concentrated and mixed with a matrix gel material wherein said chondrocytes in said matrix gel exhibit increased production of extracellular matrix material.

24. A scaffold comprising the matrix gel of claim 26.