Methods and compositions for the repair of articular cartilage defects in mammals

• US 5,723,331 A
• Filed: 06/06/1995
• Issued: 03/03/1998
• Est. Priority Date: 05/05/1994
• Status: Expired due to Fees

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Tubo, Ross A., Barone, Leesa M., Wrenn, Courtney A.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US08/469,966
• Time in Patent Office: 1,001 Days
• Field of Search: 435/240.2, 435/325, 435/366, 435/395, 435/397, 424/423, 514/2, 514/8, 623/11
• US Class Current: 435/366
• CPC Class Codes: A61B 17/00491 Surgical glue applicators surgical adhesives A61L24/00; two-component delivery syringes A61M5/19
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2/3094 Designing or manufacturing processes
• CPC Class Codes: A61F 2002/2817 Bone stimulation by chemical reactions or by osteogenic or biological products for enhancing ossification, e.g. by bone morphogenetic or morphogenic proteins [BMP] or by transforming growth factors [TGF]
• CPC Class Codes: A61F 2002/30125 elliptical or oval
• CPC Class Codes: A61F 2002/30762 Means for culturing cartilage
• CPC Class Codes: A61F 2002/30764 Cartilage harvest sites
• CPC Class Codes: A61F 2002/30971 Laminates, i.e. layered products
• CPC Class Codes: A61F 2230/0008 elliptical or oval
• CPC Class Codes: A61F 2240/001 Designing or manufacturing processes
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• 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 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3654 Cartilage, e.g. meniscus
• 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: C12N 5/0655 Chondrocytes; Cartilage

Abstract

Provided are methods and compositions for the repair of articular cartilage defects in a mammal. Denuded chondrogenic cells are proliferated ex vivo as monolayer cultures in order to expand the pool of available chondrogenic cells. During proliferation the chondrogenic cells stop secreting the extracellular matrix components, type II collagen and sulfated proteoglycans. The proliferated cells then are seeded into a pre-shaped well having a cell contacting, cell abhesive surface. The cells cultured in the well redifferentiate and begin to secrete cartilage-specific extracellular matrix again. Accordingly, essentially unlimited amounts of synthetic cartilage may be prepared from small samples of biopsy tissue. Also provided are methods for surgically repairing articular cartilage defects in mammals using the synthetic cartilage prepared in accordance with the invention.

Claims

1. A method for repairing an articular cartilage defect at a pre-determined site in a mammal, the method comprising the steps of:

(a) surgically implanting at the predetermined site of said defect a piece of synthetic cartilage of a predetermined volume prepared by the method comprising;

(i) seeding denuded chondrogenic cells, proliferated ex vivo, into a preshaped well having a cell contacting, cell abhesive surface, said well being dimensioned to provide a predetermined volume of the synthetic cartilage so that said volume approximates a volume of the cartilage defect to be repaired;

(ii) culturing said cells in said well for a time sufficient to permit said cells to differentiate and form a synthetic cartilage comprising chondrogenic cells dispersed within an endogenously produced extracellular matrix;

(iii) removing said synthetic cartilage from said well; and

(b) permitting the synthetic cartilage to integrate into the predetermined site.

2. The method of claim 1, comprising an additional step of fixing said synthetic cartilage at said pre-determined site.

3. The method of claim 2, wherein said additional step comprises surgically fixing said synthetic cartilage at said pre-determined site.

4. The method of claim 2, wherein said additional step comprises applying a bioadhesive to the interface of said pre-determined site and said synthetic cartilage.

5. The method of claim 1, wherein said method comprises the additional step of excising defective articular cartilage from said site prior to implanting the synthetic cartilage.

6. The method of claim 1, wherein step (a) is effected by implanting a plurality of synthetic cartilage pieces at said site.

7. The method of claim 1, wherein said defect is a partial thickness defect.

8. The method of claim 1, wherein said synthetic cartilage is autogenic or allogeneic.

11. The method of claims 1, 9 or 10, wherein said cell abhesive surface comprises a surface coated with silicon or polytetrafluoroethylene.

12. The method of claims 1, 9 or 10, wherein said well is defined by a material selected from the group consisting of agarose, glass, untreated cell culture plastic and polytetrafluoroethylene.

13. The method of claims 1, 9 or 10, wherein said chondrogenic cells are derived from cartilage.

14. The method of claims 1, 9 or 10, further comprising an additional step of adding a polypeptide growth factor selected from the group consisting of transforming growth factor -β

, platelet derived growth factor, insulin-like growth factor, acidic fibroblast growth factor, basic fibroblast growth factor, epidermal growth factor, hepatocytic growth factor, keratinocyte growth factor, and bone morphogenic protein to the cells prepared by the method in step (a).

9. A method for repairing an articular cartilage defect at a pre-determined site in a mammal, the method comprising the steps of:

(a) surgically implanting at the predetermined site of said defect a piece of synthetic cartilage of a pre-determined volume prepared by the method comprising;

(i) providing a tissue comprising interconnected chondrogenic cells;

(ii) disaggregating said tissue to release denuded cells;

(iii) proliferating said denuded cells ex vivo;

(iv) seeding said proliferated cells into a preshaped well having a cell contacting, cell abhesive surface, said well being dismensioned to provide a predetermined volume of the synthetic cartilage so that said volume approximates a volume of the cartilage defect to be repaired;

(v) culturing said cells in said well for a time sufficient to permit said cells to differentiate and form a formation of synthetic cartilage comprising chondrogenic cells dispersed within an endogenously produced extracellular matrix;

(vi) removing said synthetic cartilage from said well; and

(b) permitting the synthetic cartilage to integrate into the pre-determined site.

10. A method for repairing an articular cartilage defect at a predetermined site in a mammal, the method comprising the steps of:

(a) surgically implanting at the predetermined site of said defect a piece of synthetic cartilage of a pre-determined volume prepared by the method comprising;

(i) seeding denuded chondrogenic cells, proliferated ex vivo, into a preshaped well having a cell contacting, cell abhesive surface, said well being dimensioned to provide a pre-determined volume of the synthetic cartilage so that said volume approximates a volume of the cartilage defect to be repaired;

(ii) adding ascorbate to said cells in said well;

(iii) culturing said cells in said well for a time sufficient to permit said cells to differentiate and form a synthetic cartilage comprising chondrogenic cells dispersed within an endogenously produced extracellular matrix;

(iv) removing said synthetic cartilage from said well; and

(b) permitting the synthetic cartilage to integrate into the predetermimined site.

15. A method for repairing an articular cartilage defect at a pre-determined site in a mammal, the method comprising the steps of:

(a) surgically implanting at the predetermined site of said defect a piece of synthetic cartilage of a predetermined volume prepared by the method consisting of;

(i) seeding denuded chondrogenic cells, proliferated ex vivo, into a preshaped well having a cell contacting, cell abhesive surface, said well being dimensioned to provide a predetermined volume of the synthetic cartilage so that said volume approximates a volume of the cartilage defect to be repaired;

(ii) culturing said cells in said well for a time sufficient to permit said cells to differentiate and form a synthetic cartilage consisting of chondrogenic cells dispersed within an endogenously produced extracellular matrix;

(iii) removing said synthetic cartilage from said well; and

(b) permitting the synthetic cartilage to integrate into the predetermined site.