Method and material for enhanced tissue-biomaterial integration

• US 20040170663A1
• Filed: 10/09/2003
• Published: 09/02/2004
• Est. Priority Date: 10/09/2002
• Status: Active Grant

• Current Assignee: Synthasome, Inc.
• Original Assignee: Jennifer H. Elisseeff
• Inventors: Ratcliffe, Anthony, Elisseeff, Jennifer H., Wang, Dongan
• Granted Patent: US 7,862,831 B2
• Time in Patent Office: Days
• US Class Current: 424/423
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61L 24/0005 Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: A61L 24/0031 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3616 Blood, e.g. platelet-rich plasma
• CPC Class Codes: A61L 27/3633 Extracellular matrix [ECM]
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

The present invention relates to the covalent binding of a hydrogel to an extracellular matrix (ECM). The integration of the hydrogel with the tissue is superior to that in previous techniques. Moreover, unlike previous techniques, the present invention does not require a photoinitiator. Potential therapeutic applications include tissue repair and delivery of drugs or cells.

The ECM is first exposed, then treated with a priming agent. Then a polymerizable agent is added and crosslinked to the ECM.

Two primary embodiments of methods are disclosed. In the first, the priming agent is an oxidizer which creates tyrosyl radicals in the ECM, which are then bound by acrylate groups in the polymerizable agent. In the second, the priming agent contains aldehydes which bind amino groups in the ECM.

Claims

1. A method of integrating a hydrogel to an extracellular matrix, comprising the steps of:

(a) providing an exposed extracellular matrix;

(b) priming the extracellular matrix by treating with a priming agent to create a primed extracellular matrix;

(c) adding to the primed extracellular matrix a solution of a polymerizable agent; and

(d) reacting the primed extracellular matrix and polymerizable agent to create a hydrogel covalently bound to the extracellular matrix.

2. The method of claim 1, wherein:

said extracellular matrix comprises a plurality of tyrosine residues,

said priming step comprises oxidizing said extracellular matrix,

said priming agent comprises an oxidizing agent,

said primed extracellular matrix comprises a plurality of tyrosyl radicals,

said polymerizable agent comprises an acrylate group reactable with said tyrosyl radicals, and

said reacting step comprises binding said polymerizable agent to the tyrosyl radicals and crosslinking said polymerizable agent.

3. The method of claim 2, wherein said extracellular matrix is disposed in a living body.

4. The method of claim 3, wherein said body is that of a mammal.

5. The method of claim 4, wherein said body is that of a human.

6. The method of claim 3, wherein said providing step comprises treating said extracellular matrix with chondroitinase ABC.

7. The method of claim 2, wherein said extracellular matrix comprises a collagen.

8. The method of claim 7, wherein said collagen is disposed in a cartilage.

9. The method of claim 2, wherein said oxidizing agent comprises a hydrogen peroxide solution.

10. The method of claim 2, wherein said priming step further includes exposing said extracellular matrix and said polymerizable agent to a source of electromagnetic radiation.

11. The method of claim 2, wherein said source of electromagnetic radiation is a source of ultraviolet light.

12. The method of claim 2, wherein said polymerizable agent has at least one free radical polymerizable group selected from the group consisting of an of an acrylate, a diacrylate, oligoacrylate, dimethacrylate, and oligomethacrylate.

13. The method of claim 2, wherein said reacting step comprises exposing said extracellular matrix and said polymerizable agent to a source of ultraviolet radiation.

14. The method of claim 2, wherein said reacting step comprises using a redox initiator.

15. The method of claim 2, wherein said reacting step comprises an ionic crosslinking reaction.

16. The method of claim 2, wherein said reacting step comprises an enzymatic crosslinking reaction.

17. The method of claim 2, wherein said method is carried out transdermally.

18. The method of claim 2, wherein a diffusible drug substance is added with said polymerizable agent.

19. The method of claim 2, wherein a plurality of living cells are added with said polymerizable agent.

20. The method of claim 2, whereby a solid repair in a tissue is created.

25. The method of claim 1, wherein:

said extracellular matrix comprises a plurality of amino groups,

said priming agent comprises a compound with an aldehyde group and a carrier compound with an amino group,

said priming step comprises reacting said priming agent with said extracellular matrix to covalently bind said extracellular matrix and said priming agent,

said primed extracellular matrix consists of said extracellular matrix covalently bound to said priming agent, and

said reacting step consists of covalently binding said polymerizable agent to said primed extracellular matrix and crosslinking said polymerizable agent.

26. The method of claim 25, wherein said extracellular matrix is disposed in a living body.

27. The method of claim 26, wherein said body is that of a mammal.

28. The method of claim 27, wherein said body is that of a human.

29. The method of claim 25, wherein said polymerizable agent has at least one free radical polymerizable group selected from the group consisting of an of an acrylate, a diacrylate, oligoacrylate, dimethacrylate, and oligomethacrylate.

30. The method of claim 25, wherein said reacting step comprises exposing said extracellular matrix and said polymerizable agent to a source of ultraviolet radiation.

31. The method of claim 25, wherein said reacting step comprises using a redox initiator.

32. The method of claim 25, wherein said reacting step comprises an ionic crosslinking reaction.

33. The method of claim 25, wherein said reacting step comprises an enzymatic crosslinking reaction.

34. The method of claim 25, wherein said method is carried out transdermally.

35. The method of claim 25, wherein a diffusible drug substance is added with said polymerizable agent.

36. The method of claim 25, wherein a plurality of living cells are added with said polymerizable agent.

37. The method of claim 25, whereby a solid repair in a tissue is created.

21. A hydrogel produced by the method comprising the following steps:

(e) providing an exposed extracellular matrix;

(f) priming the extracellular matrix by treating with a priming agent to create a primed extracellular matrix;

(g) adding to the primed extracellular matrix a solution of a polymerizable agent; and

(h) reacting the primed extracellular matrix and polymerizable agent to create a hydrogel covalently bound to the extracellular matrix;

wherein

said extracellular matrix comprises a plurality of tyrosine residues, said priming step comprises oxidizing said extracellular matrix, said priming agent comprises an oxidizing agent, said primed extracellular matrix comprises a plurality of tyrosyl radicals, said polymerizable agent comprises an acrylate group reactable with said tyrosyl radicals, and said reacting step comprises binding said polymerizable agent to the tyrosyl radicals and crosslinking said polymerizable agent.

22. A hydrogel covalently bound to an extracellular matrix at a plurality of tyrosine residues of said matrix.

23. A hydrogel in accordance with claim 22, wherein said extracellular matrix is composed of a plurality of collagen fibers in a cartilage matrix.

24. A method of forming a hydrogel covalently bound to a cartilage tissue, comprising the steps of:

(a) exposing a plurality of tyrosine residues disposed in a plurality of collagen fibers further disposed in a cartilage tissue by treating the tissue with a glycanase to remove a plurality of polysaccharides;

(b) oxidizing the tyrosine residues, by treating with a hydrogen peroxide solution and a source of ultraviolet light, to produce a plurality of tyrosyl groups in the collagen;

(c) adding a polymerizable agent comprising one or more acrylate groups; and

(d) reacting the tyrosyl groups and the polymerizable agent to create a hydrogel covalently bound to the collagen.

38. A hydrogel produced by the method comprising the following steps:

(e) providing an exposed extracellular matrix;

(f) priming the extracellular matrix by treating with a priming agent to create a primed extracellular matrix;

(g) adding to the primed extracellular matrix a solution of a polymerizable agent; and

(h) reacting the primed extracellular matrix and polymerizable agent to create a hydrogel covalently bound to the extracellular matrix;

wherein,

said extracellular matrix comprises a plurality of amino groups, said priming agent comprises a compound with an aldehyde group and a carrier compound with an amino group, said priming step comprises reacting said priming agent with said extracellular matrix to covalently bind said extracellular matrix and said priming agent, said primed extracellular matrix consists of said extracellular matrix covalently bound to said priming agent, and said reacting step consists of covalently binding said polymerizable agent to said primed extracellular matrix and crosslinking said polymerizable agent.

39. A hydrogel covalently bound via a priming agent to an extracellular matrix at a plurality of amino groups in said extracellular matrix.