Material and Methods for Nerve Grafting

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.

1 . A method for preparing a nerve graft for implantation comprising applying at least one chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft. 
     
     
         2 . The method according to  claim 1 , wherein the chondroitin sulfate proteoglycan-degrading enzyme is selected from the group consisting of chondroitinase, hyalurondiase, 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 - 9 . (canceled) 
     
     
         10 . 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. 
     
     
         11 - 23 . (canceled) 
     
     
         24 . 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 a damaged nerve. 
     
     
         25 . 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 a damaged nerve. 
     
     
         26 . The method according to  claim 1 , wherein the nerve graft comprises peripheral nerve tissue. 
     
     
         27 . 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. 
     
     
         28 . 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. 
     
     
         29 - 33 . (canceled) 
     
     
         34 . The method according to  claim 1 , wherein the nerve graft is within the range of about 1 centimeter to about 10 centimeters in length. 
     
     
         35 . The method according to  claim 1 , wherein the nerve graft is greater than about 10 centimeters in length. 
     
     
         36 . The method according to  claim 1 , wherein the nerve graft is a living nerve graft. 
     
     
         37 . The method according to  claim 1 , wherein said nerve graft is a living graft and wherein said method further comprises rendering the living nerve graft acellular. 
     
     
         38 . The method according to  claim 37 , wherein the nerve graft is a living graft, and wherein the living nerve graft is rendered acellular before applying the chondroitin sulfate proteoglycan-degrading enzyme to the nerve graft. 
     
     
         39 . The method according to  claim 1 , wherein the nerve graft is a living graft, and wherein said method further comprises rendering the living nerve graft acellular by freeze-killing the nerve graft. 
     
     
         40 . The method according to  claim 1 , wherein the nerve graft is a living graft, and wherein said method further comprises rendering the living nerve graft acellular by chemical extraction with detergents. 
     
     
         41 . The method according to  claim 1 , wherein the nerve graft is derived from a mammal. 
     
     
         42 - 143 . (canceled)