Cell seeding of ceramic compositions

• US 6,544,290 B1
• Filed: 02/13/1998
• Issued: 04/08/2003
• Est. Priority Date: 05/19/1995
• Status: Expired due to Fees

• Current Assignee: Life Science Enterprises , INC.
• Original Assignee: Etex Corporation (Zimmer Biomet Holdings, Inc.)
• Inventors: Rey, Christian, Aiolova, Maria, Lee, Dosuk D.
• Primary Examiner(s): Isabella, David J.
• Application Number:
  US09/023,340
• Time in Patent Office: 1,880 Days
• Field of Search: 623/11.11, 623/16.11, 623/23.61, 623/23.62, 623/23.63, 424/602, 424/422, 424/426, 428/404, 423/308, 433/180, 433/201.1
• US Class Current: 623/23.63
• CPC Class Codes: A61F 2/28 Bones joints A61F2/30
• 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 2002/30762 Means for culturing cartilage
• CPC Class Codes: A61F 2002/4648 Means for culturing bone graft
• CPC Class Codes: A61F 2210/0004 bioabsorbable
• CPC Class Codes: A61F 2310/00293 containing a phosphorus-containing compound, e.g. apatite
• CPC Class Codes: A61K 9/2009 Inorganic compounds
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/426 Immunomodulating agents, i.e. cytokines, interleukins, interferons
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 24/02 containing inorganic materials
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 27/12 Phosphorus-containing materials, e.g. apatite
• CPC Class Codes: A61L 27/3821 Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
• CPC Class Codes: A61L 27/3847 Bones
• CPC Class Codes: A61L 27/3852 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/425 of phosphorus containing material, e.g. apatite
• CPC Class Codes: A61L 27/54 : Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/58 : Materials at least partially resorbable by the body
• CPC Class Codes: C01B 25/16 : Oxyacids of phosphorus; Salts thereof peroxyacids or salts thereof C01B15/00
• CPC Class Codes: C01B 25/32 : Phosphates of magnesium, calcium, strontium, or barium
• CPC Class Codes: C01B 25/321 : Methods for converting an alkaline earth metal ortho-phosphate into another ortho-phosphate by reaction, e.g. of phosphate rock with phosphoric acid C01B25/322
• CPC Class Codes: C01B 25/325 : Preparation by double decomposition
• CPC Class Codes: C04B 2111/00836 : for medical or dental applications
• CPC Class Codes: C04B 28/344 : the phosphate binder being present in the starting composition solely as one or more phosphates
• CPC Class Codes: Y10S 623/919 : Bone

Abstract

The present invention provides a synthetic, poorly-crystalline apatitic (PCA) calcium phosphate material seeded with cells. Preferably, the cells are tissue-forming or tissue-degrading cells. The compositions provided by the present invention are useful for a variety of applications, including in vivo and in vitro tissue growth (preferably bone or cartilage), osseous augmentation, and methods of diagnosing disease states by assaying tissue-forming potential of cells isolated from a host. The invention also provides in vitro cell culture systems and cell encapsulation matrices.

Claims

1. A cell-scaffold composition prepared in vitro for use as a biocompatible implant, comprising:

i) a three-dimensional scaffold comprising a hydrated precursor that comprises amorphous calcium phosphate that is capable of being converted into a synthetic poorly crystalline apatitic calcium phosphate; and

ii) at least one cell attached in vitro to the surface of the scaffold.

2. The implant of claim 1, wherein said implant further comprises a promoter.

3. The implant of claim 1, wherein the at least one cell is selected from the group consisting of chondrocytes, osteocytes, osteoblasts, osteoclasts, mesenchymal stem cells, fibroblasts, muscle cells, hepatocytes, parenchymal cells, cells of intestinal origin, nerve cells, and skin cells.

4. The implant of claim 1, wherein the at least one cell comprises at least one tissue-forming cell.

5. The implant of claim 4 wherein the at least one cell comprises at least one bone-forming cell.

6. The implant of claim 4 wherein the at least one cell comprises at least one cartilage-forming cell.

7. The implant of claim 1, wherein the at least one cell comprises a tissue-degrading cell.

8. The implant of any one of claims 3-7 or 1, wherein the at least one cell comprises a preparation selected from the group consisting of primary tissue explants, preparations of primary tissue explants, isolated cells, cell lines, transformed cell lines, and host cells.

9. The implant of claim 1, wherein the PCA calcium phosphate is strongly resorbable.

10. The implant of claim 9 wherein the PCA calcium phosphate has a X-ray diffraction pattern substantially as shown in FIG. 5d.

11. The implant of claim 9 wherein the PCA calcium phosphate has an X-ray diffraction pattern comprising broad peaks at 2θ

values of 26°

, 28.5°

, 32°

, and 33°

.

12. The implant of claim 9 wherein the PCA calcium phosphate has a calcium to phosphate ratio of less than about 1.5.

13. The implant of claim 9, wherein the hydrated precurser is a paste.

14. The implant of claim 13, wherein the hydrated precurser is characterized by a tendency to harden at 22°

C. after a time longer than one hour.

15. The implant of claim 13, wherein the hydrated precurser is characterized by a tendency to harden at 37°

C. after a time shorter than one hour.

16. The implant of claim 13, wherein the hydrated precurser is characterized by a tendency to harden at 22°

C. after about 10-30 minutes.

17. The implant of claim 13, wherein the hydrated precursor is characterized in that it substantially hardens within about 15-40 minutes when placed in vivo.

18. The implant of claim 1, further comprising an additional material selected to change a physical parameter of the implant, which physical parameter is selected from the group consisting of:

strength, resorption time, adherence, injectability, frictional characteristics, and release kinetics.

19. The implant of claim 1, further comprising a biologically active agent.

20. The implant of claim 19, wherein the biologically active agent is one that affects a process selected from the group consisting of cell growth, cell migration, cell differentiation, and cell localization.

21. The implant of claim 19, wherein the biologically active agent is selected from the group consisting of growth factors and extracellular matrix components.

22. The implant of claim 19, wherein the biologically active agent is selected from the group consisting of lamin, fibronectin, collagen, and combinations thereof.

23. The implant of claim 19, wherein the biologically active agent is selected from the group consisting of nutrients, angiogenic factors, and immunomodulatory factors.

24. The implant of claim 2, wherein the promoter comprises a second calcium phosphate that participates in the conversion reaction and is selected to have appropriates stoichiometry so that the resultant PCA has a Ca/P ratio within the range of 1.1-1.9.

25. The implant of claim 24 wherein the second calcium phosphate is selected from the group consisting of calcium metaphosphate, dicalcium phosphate dehydrate, heptacalcium decaphosphate, tricalcium phosphates, calcium pyrophosphate dehydrate, PCA calcium phosphate, calcium pyrophosphate, octacalcium phosphate, tetracalcium phosphate, and amorphous calcium phosphates.

26. The vehicle of claim 24 wherein the promoter is DCPD.

27. The vehicle of claim 24 wherein the promoter is stoichiometric hydroxyapatite.

28. The implant of claim 24 wherein the promoter is selected from the group consisting of calcium sources, phosphate sources, and combinations thereof, the promoter being one that participates in the reaction and being selected for appropriate stoichiometry so that the resultant PCA calcium phosphate has a Ca/P ration in the range of approximately 1.1-1.9.

29. The implant of claim 2, wherein promoter does not participate in the conversion reaction.

30. The implant of claim 29 wherein the promoter is selected from the group consisting of Al₂O₃, mica, glass, sand, and heat.

31. The implant of claim 23 wherein the promoter is a granular material having an average grain diameter of approximately 1-200 μ

m.

32. The vehicle of claim 23 wherein the promoter is bioresorbable.