Neomorphogenesis of cartilage in vivo from cell culture

• US 5,041,138 A
• Filed: 04/17/1989
• Issued: 08/20/1991
• Est. Priority Date: 11/20/1986
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Hospital, Massachusetts Institute of Technology
• Inventors: Langer, Robert S., Vacanti, Joseph P., Vacanti, Charles A.
• Primary Examiner(s): NOT, DEFINED
• Application Number:
  US07/339,155
• Time in Patent Office: 855 Days
• Field of Search: 623/16, 623/66, 623/11, 623/15, 623/10, 623/18, 424/548, 514/21, 514/801, 128/DIG. 8, 530/840, 435/240.2
• US Class Current: 424/422
• CPC Class Codes: A61F 2/02 Prostheses implantable into the body closure means for urethra or rectum or for artificial body openings therefor A61F2/0004
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61F 2/30 Joints
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2002/30062 (bio)absorbable, biodegradable, bioerodable, (bio)resorbable, resorptive
• CPC Class Codes: A61F 2210/0004 bioabsorbable
• 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 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: C12N 2533/18 Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• 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
• CPC Class Codes: Y10S 530/84 Bones; tendons; teeth; cartilage
• CPC Class Codes: Y10S 623/919 Bone

Abstract

Methods and artificial matrices for the growth and implantation of cartilaginous structures and surfaces are disclosed. In the preferred embodiments, chondrocytes are grown on biodegradable, biocompatible fibrous polymeric matrices. Optionally, the cells are proliferated in vitro until an adequate cell volume and density has developed for the cells to survive and proliferate in vivo. One advantage of the matrices is that they can be cast or molded into a desired shape, on an individual basis, so that the final product closely resembles a patient'"'"'s own ear or nose. Alternatively, flexible matrices can be used which can be manipulated at the time of implantation, as in a joint, followed by remodeling through cell growth and proliferation in vivo. The cultured cells can also be maintained on the matrix in a nutrient media for production of bioactive molecules such as angiogenesis inhibiting factor.

Claims

1. A method for making a cartilaginous structure comprising

providing a biocompatible, biodegradable synthetic polymeric matrix in a nutrient environment and

attaching cartilage cells to the matrix to form a cartilaginous structure suitable for implantation into a patient to replace defective or missing cartilage.

2. The method of claim 10 wherein the polymer is selected from the group consisting of on histologic evaluation. In vitro, all polymer fibers (controls and experimentals) began to dissolve by day 27. On gross and histologic examination using Hematoxylin and Eosin stain, none of the 16 specimens designated as controls displayed any evidence of cartilage. In contrast, 18 of 20 specimens in the experimental group showed evidence of cartilage formation grossly, as well as histologically using Hematoxylin and Eosin stain. Histologic examination of the implants removed at day 8 showed the fibers were imbedded in fibrous tissue with evidence of a mild inflammatory response consisting of infiltrates of polymorphonuclear leukocytes and giant cells, and isolated "nests" of cartilage. During the time intervals to day 18 and day 28, these islands of cartilage grew and coalesced into a large homogenous mass of cartilage. There was no evidence of neovascularization in the 49- and 81- day implants, and there was decreasing evidence of an inflammatory response with time as characterized by a decrease in the number of polymorphonuclear leukocytes and giant cells. Very little evidence of the polymer fibers was seen after 28 days. This increase in the size of the cartilage appeared to be at the expense of the fibrous tissue previously seen and associated at least temporarily with a decrease in neovascularization and resolution of the mild inflammatory response originally noted. Also associated with this was the absorption of the biodegradable polymer fibers. In time, the polymer fibers were progressively replaced by

3. The method of claim 1 further comprising coating the polymer with a material selected from the group consisting of basement membrane components, agar, agarose, gelatin, gum arabic, collagens, fibronectin, laminin, glycosaminoglycans, attachment septides, functional equivalents and mixtures thereof.

4. The method of claim 1 further comprising providing cells selected from the group consisting of bone, skin and nerve cells.

5. The method of claim 1 wherein the matrix is formed as a rigid structure.

6. The method of claim 1 wherein the matrix is formed as a flexible structure conformable to a joint surface.

7. The method of claim 1 wherein the nutrient environment is in vivo further comprising attaching the cells on a matrix and implanting the attached cells on the matrix in an animal without first proliferating the cells on the matrix in vitro.

8. The method of claim 1 further comprising attaching the cells on a matrix, proliferating the cells on the matrix in vitro in a nutrient media, then implanting the cells on the matrix in vitro.

9. The method of claim 1 further comprising implanting the structure as part of an ear.

10. The method of claim 1 further comprising implanting the structure as part of a nose.

11. The method of claim 1 further comprising implanting the structure on the surface of a joint.

12. The method of claim 1 for producing bioactive molecules in vitro further comprising culturing the cells on the matrix in vitro in a nutrient medium until bioactive molecule is produced.

13. The method of claim 12 further comprising extracting the bioactive molecule from the nutrient medium.

14. The method of claim 12 further comprising extracting the bioactive molecule from the cells on the matrix.

15. The method of claim 1 further comprising providing a substance in the nutrient environment or polymer and determining if the substance has an effect on the chondrocyte cells.

16. The method of claim 1 further comprising characterizing the effect of the substance on the chondrocyte cells.

17. The method of claim 1 further comprising providing cells capable of differentiating into cartilage cells and inducing differentiation into cartilage cells.

18. The method of claim 4 wherein the cartilage cells are induced to form bone cells.

19. The method of claim 1 wherein the structure is implanted to repair cartilage damaged by inflammation.

20. The method of claim 1 wherein the structure is implanted to repair cartilage damaged by trauma.

21. The method of claim 1 wherein the structure is implanted to repair cartilage damaged by aging.

22. The method of claim 1 wherein the structure is implanted to repair cartilage when is congenitally defective.