Cell-support matrix having narrowly defined uniformly vertically and non-randomly organized porosity and pore density and a method for preparation thereof

• US 9,701,940 B2
• Filed: 12/19/2014
• Issued: 07/11/2017
• Est. Priority Date: 09/19/2005
• Status: Active Grant

• Current Assignee: Ocugen, Inc.
• Original Assignee: Histogenics Corporation (Ocugen, Inc.)
• Inventors: Kusanagi, Akihiko, Claesson, Hans P. I., Tarrant, Laurence J. B., Smith, R. Lane
• Primary Examiner(s): Tate, Chris R
• Assistant Examiner(s): Kosar, Aaron J
• Application Number:
  US14/577,610
• Publication Number:
  US 20150104872A1
• Time in Patent Office: 935 Days
• Field of Search: None
• CPC Class Codes: A61F 2/08 Muscles; Tendons; Ligaments sutures A61B17/04
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2/3872 Meniscus for implantation between the natural bone surfaces
• CPC Class Codes: A61F 2002/30062 (bio)absorbable, biodegradable, bioerodable, (bio)resorbable, resorptive
• CPC Class Codes: A61F 2210/0004 bioabsorbable
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61L 2400/18 Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
• CPC Class Codes: A61L 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 2430/10 for reconstruction of tendons or ligaments
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/3817 Cartilage-forming cells, e.g. pre-chondrocytes
• CPC Class Codes: A61L 27/3843 Connective tissue
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2535/00 Supports or coatings for cell culture characterised by topography
• 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 cell-support matrix having narrowly defined uniformly vertically and non-randomly organized porosity and pore density and a method for preparation thereof. The matrix suitable for preparation of cellular or acellular implants for growth and de novo formation of an articular hyaline-like cartilage. A gel-matrix composite system comprising collagen-based matrix having a narrowly defined porosity capable of inducing hyaline-like cartilage production from chondrocytes in vivo and in vitro.

Claims

1. An implant comprising:

a collagen scaffold comprising pores, having a basal surface and an apical surface each with openings to the pores, wherein at least 85% of the pores in the scaffold are vertically oriented; and

a suspension of chondrocytes seeded into the scaffold.

2. The implant of claim 1, wherein at least 85% of the openings of the apical surface have a diameter of 200±

100 micrometers.

3. The implant of claim 2, wherein the apical surface has a pore density of 25±

10 pores per mm².

4. The implant of claim 1, wherein at least 85% of the openings of the basal surface have a diameter of 200±

100 micrometers.

5. The implant of claim 4, wherein the basal surface has a pore density of 25±

10 pores per mm².

6. The implant of claim 1, wherein at least 95% of the basal and apical openings have diameters of 200±

100 micrometers.

7. The implant of claim 6, wherein at least 98% of said basal and apical openings have diameters of 200±

100 micrometers.

8. The implant of claim 1, wherein the collagen scaffold comprises Type I collagen, Type II collagen, Type IV collagen or a combination thereof.

9. The implant of claim 1, wherein the scaffold is formed by polymerizing a collagen solution in the presence of ammonia vapor.

10. The implant of claim 9, wherein the collagen solution is polymerized at a reduced pressure.

11. The implant of claim 10, wherein the collagen solution is polymerized at between 3 Torr and 10 Torr of pressure.

12. The implant of claim 9, wherein the collagen solution comprises between 4 mg/ml and 8 mg/ml of Type I collagen in aqueous solution.

13. The implant of claim 12, wherein the aqueous solution is acidic.

14. The implant of claim 12, wherein the collagen solution comprises a non-ionic surfactant.

15. The implant of claim 14, wherein the non-ionic surfactant comprises a polyoxyethylene-polyoxypropylene block copolymer.

16. The implant of claim 9, further comprising freeze-drying the scaffold prior to seeding the scaffold with chondrocytes.

17. The implant of claim 1, wherein the chondrocytes are seeded as a suspension of chondrocytes in a collagen solution.

18. The implant of claim 17, wherein the collagen chondrocyte suspension has a concentration of 1 million to 12 million chondrocyte cells per mL of collagen solution.

19. The implant of claim 1, wherein the chondrocytes comprise human articular chondrocytes.