Method of obtaining viable small tissue particles and use for tissue repair

• US 8,497,121 B2
• Filed: 03/23/2012
• Issued: 07/30/2013
• Est. Priority Date: 12/20/2006
• Status: Expired due to Fees

• Current Assignee: Zimmer Orthobiologics, Inc. (Zimmer Biomet Holdings, Inc.)
• Original Assignee: Zimmer Orthobiologics, Inc. (Zimmer Biomet Holdings, Inc.)
• Inventors: Yao, Jian Q., Zaporojan, Victor
• Primary Examiner(s): Ariani, Kade
• Application Number:
  US13/428,873
• Publication Number:
  US 20120183586A1
• Time in Patent Office: 494 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 38/4833 Thrombin (3.4.21.5)
• CPC Class Codes: A61K 9/14 Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles microspheres A61K9/16; microcapsules A61K9/50; nanocapsules, nanoparticles of the matrix type A61K9/51
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue

Abstract

The invention provides a composition including isolated small living tissue particles, a method of making the tissue particles, and a method of using the composition to ameliorate a tissue defect. The tissue particles are composed of cells and their associated extracellular molecules and are sized, in certain embodiments, to be smaller than about 1 mm. Another aspect of the inventive tissue particles is the large percentage of viable cells. In certain embodiments, the tissue particles are made from cartilage and the composition may also contain additives such as adhesives, solutions, and bioactive agents.

Claims

1. A composition comprising:

a plurality of isolated allogenic human juvenile cartilage particles, the cartilage particles including chondrocytes, at least about 50% of which are viable, and chondrocyte-associated extracellular biomolecules, each cartilage particle having at least one dimension less than 1 mm; and

a cartilage particle maintenance solution.

2. The composition of claim 1, wherein the cartilage particles are obtained from a juvenile donor less than 12 years of age.

3. The composition of claim 1, wherein the cartilage particles comprise articular cartilage.

4. The composition of claim 1, wherein the percentage of viable chondrocytes is selected from at least about 60%, at least about 65%, at least about 70%, at least about 75% and at least about 80%.

5. The composition of claim 4, wherein the percentage of viable chondrocytes is at least about 75%.

6. The composition of claim 1, wherein the percentage of viable chondrocytes is determined by a membrane permeability assay.

7. The composition of claim 6, wherein the membrane permeability assay includes fluorescent labeling of nucleic acids.

8. The composition of claim 1, wherein the chondrocyte-associated extracellular biomolecules are selected from proteins, polysaccharides, proteoglycans and combinations thereof.

9. The composition of claim 8, wherein the chondrocyte-associated extracellular biomolecules are combinations of proteins, polysaccharides and proteoglycans.

10. The composition of claim 1, wherein the cartilage particles have a volume less than 1 mm³.

11. The composition of claim 10, wherein the cartilage particles have a volume of about 2×

10<sup>−

4</sup> mm³.

12. The composition of claim 1, wherein the maintenance solution comprises a buffer.

13. The composition of claim 12, wherein the buffer maintains the solution pH between about 5 and about 7.

14. The composition of claim 13, wherein the buffer maintains the solution pH between about 6.8 and about 7.5.

15. The composition of claim 1, further comprising at least one bioactive agent.

16. The composition of claim 15, wherein the at least one bioactive agent is selected from a growth factor, a hormone, a nutrient and combinations thereof.

17. The composition of claim 16, wherein the growth factor is selected from a BMP, an IGF, a TGF, a PDGF and combinations thereof.

18. The composition of claim 17, wherein the growth factor is a TGF.

19. The composition of claim 1, further comprising an adhesive.

20. The composition of claim 19, wherein the adhesive is selected from fibrin, hyaluronic acid, fibrin glue, fibrin clot, collagen gel, alginate gel, gelatin-resorcin-formalin adhesive, mussel-based adhesive, dihydroxyphenylalanine (DOPA) based adhesive, chitosan, transglutaminase, poly(amino acid)-based adhesive, cellulose-based adhesive, polysaccharide-based adhesive, synthetic acrylate-based adhesives, platelet rich plasma (PRP), platelet poor plasma (PPP), clot of PRP, clot of PPP, solubilized basement membrane (MATRIGEL®

), monostearoyl glycerol co-succinate (MGSA), monostearoyl glycerol co-succinate/polyethylene glycol (MGSA/PEG) copolymers, laminin, elastin, proteoglycans, and combinations thereof.

21. The composition of claim 20, wherein the adhesive includes fibrin.