Materials and methods for nerve grafting

• US 9,402,868 B2
• Filed: 02/25/2013
• Issued: 08/02/2016
• Est. Priority Date: 08/13/2001
• Status: Expired due to Term

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Muir, David F.
• Primary Examiner(s): Afremova, Vera
• Application Number:
  US13/776,606
• Publication Number:
  US 20130337549A1
• Time in Patent Office: 1,254 Days
• Field of Search: None
• US Class Current: 1/1
• 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 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• CPC Class Codes: A61K 38/51 Lyases (4)
• CPC Class Codes: A61L 2430/32 for nerve reconstruction
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• 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 27/3675 Nerve tissue, e.g. brain, spinal cord, nerves, dura mater
• 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 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3878 Nerve tissue, brain, spinal cord, nerves, dura mater
• CPC Class Codes: A61L 31/047 Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

The subject invention pertains to compositions and methods for promoting repair of damaged nerve tissue using nerve grafts and preparation of nerve grafts. The compositions and methods of the subject invention can be employed to restore the continuity of nerve interrupted by disease, traumatic events or surgical procedures. Compositions of the subject invention comprise one or more chondroitin sulfate proteoglycan (CSPG)-degrading enzymes that promote axonal penetration into damaged nerve tissue and nerve graft. The invention also concerns methods for promoting repair of damaged nerve tissue using the present compositions and nerve tissue treated according to such methods. The invention also includes storage solutions for nerve tissue.

Claims

1. A method for preparing a nerve graft for implantation into a mammal, the method comprising the steps of:

a) providing a nerve graft capable of supporting axonal ingress after implantation and comprising a structurally complete nerve segment containing an intact basal lamina tube;

b) rendering the nerve graft acellular by chemical extraction with a detergent;

c) degrading chondroitin sulfate proteoglycan of the nerve graft by applying, in vitro, at least one chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft so as to condition the nerve graft for enhanced post-implantation axonal ingress from a truncated adult peripheral nerve of the mammal into the nerve graft at a site of coaptation wherein the post-implantation axonal ingress is enhanced relative to a nerve graft to which the at least one chondroitin sulfate proteoglycan-degrading enzyme has not been applied.

2. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyaluronidase, and matrix metalloproteinase, or a combination thereof.

3. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, chondroitinase AC, hyaluronidase, matrix metalloproteinase-2, and matrix metalloproteinase-9, or a combination thereof.

4. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase ABC, chondroitinase A, chondroitinase C, and chondroitinase AC or a combination thereof.

5. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase.

6. The method according to claim 1, wherein the nerve graft has a first end and a second end, and wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied to first end of the nerve graft, or to the second end of the nerve graft, or to both the first end and second end of the nerve graft.

7. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is applied to the nerve graft by placing the nerve graft in a culture medium containing the chondroitin sulfate proteoglycan-degrading enzyme.

8. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme decreases the latency of axonal ingress into the nerve graft once the nerve graft is coapted into the transected adult peripheral nerve.

9. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme increases the rate of axonal ingress into the nerve graft once the nerve graft is coapted into the transected adult peripheral nerve.

10. The method according to claim 1, wherein the nerve graft comprises peripheral nerve tissue.

11. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase, and wherein the chondroitinase is applied to the nerve graft within a concentration range from about 10 units/mL to about 1000 units/mL.

12. The method according to claim 1, wherein the chondroitin sulfate proteoglycan-degrading enzyme is a chondroitinase, and wherein the chondroitinase is applied to the nerve graft within a concentration range from about 100 units/mL to about 500 units/mL.

13. The method according to claim 1, wherein the nerve graft is within the range of about 1 centimeter to about 10 centimeters in length.

14. The method according to claim 1, wherein the nerve graft is greater than about 10 centimeters in length.

15. The method according to claim 1, wherein the nerve graft is derived from a mammal.

16. A method for preparing a nerve graft for implantation into a mammal, the method comprising the steps of:

a) providing a nerve graft comprising a structurally complete nerve segment containing an intact basal lamina tube;

b) killing the cells in the nerve graft; and

c) degrading chondroitin sulfate proteoglycan of the nerve graft by applying, in vitro, at least one chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft so as to enhance post-implantation axonal traversal of an interface between the nerve graft and a transected adult peripheral nerve of the mammal coaptation wherein the post-implantation axonal traversal of the interface is enhanced relative to a nerve graft to which the at least one chondroitin sulfate proteoglycan-degrading enzyme has not been applied.

17. A method for preparing a nerve graft for implantation into a mammal, the method comprising the steps of:

a) providing a nerve graft capable of supporting axonal ingress after implantation and comprising a structurally complete nerve segment containing an intact basal lamina tube;

b) killing the cells in the nerve graft; and

c) degrading chondroitin sulfate proteoglycan of the nerve graft by applying, in vitro, at least one chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft so as to condition the nerve graft to allow for wider dispersal of axon sprouts from a transected adult peripheral nerve of the mammal into the basal lamina tube of the nerve graft coaptation wherein the reduced latency is enhanced relative to a nerve graft to which the at least one chondroitin sulfate proteoglycan-degrading enzyme has not been applied.

18. A method for preparing a nerve graft for implantation into a mammal, the method comprising the steps of:

a) providing a nerve graft capable of supporting axonal ingress after implantation and comprising a structurally complete nerve segment containing an intact basal lamina tube;

b) killing the cells in the nerve graft; and

c) degrading chondroitin sulfate proteoglycan of the nerve graft by applying, in vitro, at least one chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft so as to condition the nerve graft for reduced latency of post-implantation axonal ingress of a transected adult peripheral nerve of the mammal into the nerve graft coaptation wherein the reduced latency is enhanced relative to a nerve graft to which the at least one chondroitin sulfate proteoglycan-degrading enzyme has not been applied.