Patterned substrates and methods for nerve regeneration

• US 6,676,675 B2
• Filed: 04/19/2001
• Issued: 01/13/2004
• Est. Priority Date: 04/19/2000
• Status: Expired due to Fees

• Current Assignee: Iowa State University Research Foundation Incorporated (Iowa State University)
• Original Assignee: Iowa State University Research Foundation Incorporated (Iowa State University)
• Inventors: Jeftinija, Srdija, Mallapragada, Surya K., Shanks, Howard, Heath, Carole, Miller, Cheryl A.
• Primary Examiner(s): Jackson, Gary
• Application Number:
  US09/837,303
• Publication Number:
  US 20020051806A1
• Time in Patent Office: 999 Days
• Field of Search: 606/152
• US Class Current: 606/152
• 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 2430/32 for nerve reconstruction
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3821 Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
• CPC Class Codes: A61L 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61L 31/06 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds A61L31/041 takes precedence
• CPC Class Codes: A61L 31/146 Porous materials, e.g. foams or sponges
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 25/08 electrically charged
• CPC Class Codes: C12N 2502/08 cells of the nervous system
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2535/10 Patterned coating
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: Y10S 977/701 Integrated with dissimilar structures on a common substrate
• CPC Class Codes: Y10S 977/796 for electrical or electronic purpose

Abstract

Micropatterned substrates and methods for fabrication of artificial nerve regeneration conduits and methods for regenerating nerves are provided. Guidance compounds or cells are seeded in grooves formed on the patterned substrate. The substrates may also be provided with electrodes to provide electrical guidance cues to the regenerating nerve. The micropatterned substrates give physical, chemical, cellular and/or electrical guidance cues to promote nerve regeneration at the cellular level.

Claims

1. A method for regenerating a severed nerve, comprising:

providing a substrate having a surface containing one or more substantially linear grooves;

placing one or more guidance factors for nerve regeneration into said one or more grooves;

wherein said one or more guidance factors comprises poly(vinyl alcohol);

positioning said substrate in proximity to a severed end of said nerve such that said one or more grooves is substantially coextensive to said severed end; and

allowing said severed nerve to grow into said one or more grooves of said substrate.

2. The method of claim 1 wherein said substrate is in the form of a cylinder.

3. The method of claim 2 wherein said cylindrical form has an inner and outer surface and said one or more grooves are disposed on said inner surface.

4. The method of claim 1 wherein said one or more guidance factors further comprise negative guidance factors disposed on said surface in between said one or more grooves.

5. The method of claim 1, wherein said substrate comprises a material selected from the group consisting of poly(D,L-lactide), lactic acid, glycolic acids, glycolide trimethylene carbonate, polyester, polyglycolic acid, collagen, polylactic acid, poly(organo)phosphazine, polyorthoester, glycosaminoglycan, L-lactide, ε

-caprolactone, polyurethane, polyimides, and polystyrene.

6. The method of claim 1, wherein said substrate comprises poly(D,L-lactide).

7. The method of claim 1, wherein said substrate comprises copolymers of lactic and glycolic acids.

8. The method of claim 1, wherein said substrate further comprises at least one electrode.

9. The method of claim 8, wherein said at least one electrode is positioned within said one or more grooves.

10. The method of claim 1, wherein said one or more grooves are at least about 1 μ

m in width.

11. The method of claim 10, wherein said one or more grooves are up to about 50 μ

m in width.

12. The method of claim 1, wherein said one or more grooves are at least about 5 μ

m in width.

13. The method of claim 12, wherein said one or more grooves are up to about 10 μ

m in width.

14. The method of claim 1, wherein said one or more grooves are spaced at least about 10 μ

m apart.

15. The method of claim 14, wherein said one or more grooves are spaced up to about 20 μ

m apart.

16. The method of claim 14, wherein said one or more grooves are spaced up to about 100 μ

m apart.

17. The method of claim 1, wherein said one or more grooves are at least about 1 μ

m in depth.

18. The method of claim 17, wherein said one or more grooves are up to about 4 μ

m in depth.

19. The method of claim 1, wherein said one or more guidance factors comprises a material selected from the group consisting of Schwann cells, stem cells, nerve growth factor, laminin, collagen, polylysine, and chicken plasma.

20. The method of claim 1, wherein said one or more guidance factors comprises Schwann cells.

21. The method of claim 20, wherein said one or more guidance factors comprises stem cells.

22. The method of claim 21 wherein said stem cells are neuronal stem cells.

23. The method of claim 20, wherein said one or more guidance factors further comprises laminin.

24. The method of claim 23, wherein said one or more guidance factors further comprises stem cells.

25. The method of claim 24, wherein said stem cells are neuronal stem cells.

26. The method of claim 23 wherein neurite alignment of said severed nerve along said one or more grooves is at least about 90 percent.

27. The method of claim 1, wherein said one or more grooves contain Schwann cells at a concentration of at least about 50,000 cells/cm2.

28. The method of claim 27, wherein said one or more grooves contain Schwann cells at a concentration of up to about 400,000 cells/cm2.

29. The method of claim 1, wherein said one or more grooves contain laminin at a concentration of at least about 100 μ

g/ml.

30. The method of claim 29, wherein said one or more grooves contain laminin at a concentration of up to about 200 μ

g/ml.

31. The method of claim 1, wherein said severed nerve is part of the peripheral nervous system of a vertebrate.

32. The method of claim 1, wherein said severed nerve is part of the central nervous system of a vertebrate.

33. The method of claim 1, wherein said severed nerve is an optic nerve.

34. A method for regenerating a severed nerve, comprising:

providing a porous guidance conduit having an inner surface and an outer surface;

providing a substrate containing one or more substantially linear grooves, wherein said one or more grooves contain one or more guidance factors for nerve regeneration and wherein said substrate is disposed on the inner surface of said conduit;

positioning said guidance conduit in proximity to a severed end of said nerve; and

allowing said severed nerve to grow into said grooves of said substrate.

35. The method of claim 34, wherein said guidance conduit is sutured to at least one end of said severed nerve.

36. A method for regenerating a severed nerve, comprising:

providing a substrate having a surface containing a plurality of substantially linear grooves, wherein said one or more grooves are substantially parallel and contain one or more guidance factors for nerve regeneration;

wherein said substrate comprises a material selected from the group consisting of poly(D,L-lactide), and copolymers of lactic and glycolic acids;

providing a porous guidance conduit having an inner surface wherein said substrate is disposed on the inner surface of said conduit;

attaching said guidance conduit between the severed ends of said nerve; and

allowing said severed nerve to grow into said grooves of said substrate.

37. The method of claim 36 wherein said one or more guidance factors comprises a material selected from the group consisting of Schwann cells and laminin.

38. The method of claim 37 wherein said one or more grooves are at least about 5 μ

m wide, spaced at least about 10 μ

m apart, and are at least about 1 μ

m deep.

39. The method of claim 38 wherein said one or more grooves are no greater than about 10 μ

m wide, spaced no greater than about 100 μ

m apart, and are no greater than about 4 μ

m deep.

40. An apparatus for regenerating a severed nerve comprising:

a substrate having a surface containing one or more substantially linear grooves at least about 5 μ

m wide, spaced at least about 10 μ

m apart, and at least about 1 μ

m deep, wherein said one or more grooves contain one or more guidance factors for nerve regeneration, said guidance factors being selected from the group consisting of Schwann cells, stem cells, nerve growth factor, laminin, collagen, polylysine and chicken plasma; and

one or more negative guidance factors comprising poly(vinyl alchohol).

41. The apparatus of claim 40 wherein said substrate is in the form of a cylinder.

42. The apparatus of claim 40 wherein said cylindrical form has an inner and an outer surface and said one or more grooves are disposed on said inner surface.

43. The apparatus of claim 40 wherein said one or more negative guidance factors are disposed in between said one or more grooves.

44. The apparatus of claim 40, wherein said substrate comprises a material selected from the group consisting of poly(D,L-lactide), lactic acid, glycolic acids, glycolide trimethylene carbonate, polyester, polyglycolic acid, collagen, polylactic acid, poly(organo)phosphazine, polyorthoester, glycosaminoglycan, L-lactide, ε

-caprolactone, polyurethane, polyimides, and polystyrene.

45. The apparatus of claim 40, wherein said substrate comprises poly(D,L-lactide).

46. The apparatus of claim 40, wherein said substrate comprises copolymers of lactic and glycolic acids.

47. The apparatus of claim 40, wherein said substrate further comprises at least one electrode.

48. The apparatus of claim 47, wherein said electrode is positioned in said one or more grooves.

49. The apparatus of claim 40, wherein said one or more grooves are at least about 1 μ

m in width.

50. The apparatus of claim 49, wherein said one or more grooves are up to about 50 μ

m in width.

51. The apparatus of claim 40, wherein said one or more grooves are at least about 5 μ

m in width.

52. The apparatus of claim 51, wherein said one or more grooves are up to about 10 μ

m in width.

53. The apparatus of claim 40, wherein said one or more grooves are about 10 μ

m in width.

54. The apparatus of claim 40, wherein said one or more grooves are spaced at least about 10 μ

m apart.

55. The apparatus of claim 40, wherein said one or more grooves are spaced up to about 20 μ

m apart.

56. The apparatus of claim 55, wherein said one or more grooves are spaced up to about 100 μ

m apart.

57. The apparatus of claim 40, wherein said one or more grooves are at least about 1 μ

m in depth.

58. The apparatus of claim 57, wherein said one or more grooves are up to about 4 μ

m in depth.

59. The apparatus of claim 40, wherein said one or more guidance factors comprises Schwann cells.

60. The apparatus of claim 59, wherein said one or more guidance factors comprises stem cells.

61. The apparatus of claim 60, wherein said stem cells are neuronal stem cells.

62. The apparatus of claim 40, wherein said one or more guidance factors comprises laminin.

63. The apparatus of claim 62, wherein said one or more guidance factors further comprises stem cells.

64. The apparatus of claim 63, wherein said stem cells are neuronal stem cells.

65. The apparatus of claim 40, wherein said one or more grooves contain Schwann cells at a concentration of at least about 50,000 cells/cm2.

66. The apparatus of claim 40, wherein said one or more grooves contain Schwann cells at a concentration of up to about 400,000 cells/cm2.

67. The apparatus of claim 40, wherein said one or more grooves contain laminin at a concentration of at least about 100 μ

g/ml.

68. The apparatus of claim 40, wherein said one or more grooves contain laminin at a concentration of up to about 200 μ

g/ml.

69. An apparatus adapted for connection to at least one end of a severed nerve, comprising:

a cylindrical porous guidance conduit having an inner and an outer surface;

a substrate having a surface containing one or more substantially linear grooves, wherein said grooves contain one or more guidance factors for nerve regeneration; and

wherein said substrate is disposed on the inner surface of said guidance conduit.

70. An apparatus adapted for connection to at least one end of a severed nerve, comprising:

a cylindrical guidance conduit having an inner and an outer surface;

wherein said guidance conduit is sutured to at least one end of said severed nerve;

a substrate having a surface containing one or more substantially linear grooves, wherein said grooves contain one or more guidance factors for nerve regeneration; and

wherein said substrate is disposed on the inner surface of said guidance conduit.

71. An apparatus for regenerating a severed nerve, comprising:

a porous cylindrical guidance conduit having an inner and outer surface;

a substrate having a surface containing one or more substantially linear grooves, wherein said one or more grooves contain one or more guidance factors for nerve regeneration, said guidance factors being selected from the group consisting of Schwann cells, stem cells, nerve growth factor, laminin, collagen, polylysine and chicken plasma; and

wherein said substrate is disposed on the inner surface of said guidance conduit.

72. The apparatus of claim 71, wherein said substrate comprises a material selected from group consisting of poly(D,L-lactide) or copolymers of lactic and glycolic acids.

73. The apparatus of claim 72, wherein said one or more guidance factors comprises a material selected from the group consisting of Schwann cells and laminin.

74. The apparatus of claim 71 wherein said one or more grooves are at least about 5 μ

m wide, spaced at least about 10 μ

m apart, and are at least about 1 μ

m deep.

75. The apparatus of claim 74 wherein said one or more grooves are no greater than about 10 μ

m wide, spaced no greater than about 100 μ

m apart, and are no greater than about 4 μ

m deep.