Methods and Compositions for Treating Intervertebral Disc Degeneration

Abstract

A fluid matrix comprising cross-linked remodelable collagen from a donor vertebrate animal is useful for regenerating hydrodynamic function in damaged intervertebral discs in vivo. The matrix may be injectable and may comprise cells and a plurality of purified cell growth factors. The matrix promotes cell growth and elaboration of proteoglycans to facilitate regeneration of native tissues. The collagen in the matrix may be cross-linked using photooxidative catalysis and visible light, and purified cell growth factors are preferably at least partly bone-derived.

1 . A matrix for treating a patient having degenerative disc disease, the matrix comprising
 an injectable fluid comprising digestion-resistant remodelable collagen, said collagen being cross-linked through photooxidative catalysis and irradiation by visible light; and   a plurality of living cells dispersed within said injectable fluid to form an injectable cell matrix for treating degenerative disc disease, said cells having inherent capability to elaborate proteoglycans in vivo.   
     
     
         2 . The injectable matrix of  claim 1 , further comprising
 a plurality of purified cell growth factors dispersed within said injectable cell matrix to form an injectable disc regeneration fluid, said living cells being responsive to said purified cell growth factors by increased elaboration of proteoglycans in vivo.   
     
     
         3 . The matrix of  claim 1  wherein said cells are chondrocytes. 
     
     
         4 . The matrix of  claim 1  wherein said cells are mesenchymal stem cells. 
     
     
         5 . The matrix of  claim 4  wherein said cells are human-derived. 
     
     
         6 . The matrix of  claim 1  wherein said collagen is cross-linked using methylene blue as a photooxidative catalyst. 
     
     
         7 . The matrix of  claim 1  wherein said cells are cultured in vitro to increase their response to said cell growth factors. 
     
     
         8 . The injectable disc regeneration fluid of  claim 2  wherein at least two of said plurality of cell growth factors are bone-derived. 
     
     
         9 . An injectable disc regeneration fluid, comprising
 an injectable cell matrix according to  claim 7 ; and   a plurality of cell growth factors dispersed within said injectable cell matrix to form an injectable disc regeneration fluid, said living cells being responsive to said cell growth factors by increased elaboration of proteoglycans in vivo.   
     
     
         10 . A method of treating a patient presenting with degenerative disc disease, the method comprising
 providing an injectable disc regeneration fluid according to  claim 2 ; and   injecting said injectable disc regeneration fluid into at least one of said patient's intervertebral discs to treat degenerative disc disease in said disc.   
     
     
         11 . A method of continuing treatment of a patient presenting with degenerative disc disease, the method comprising
 treating the patient according to the method of  claim 10 ; and   injecting a plurality of cell growth factors into said at least one of said patient's intervertebral discs after completion of the method of  claim 10  to continue treatment of a patient presenting with degenerative disc disease.   
     
     
         12 . A patient having a history of degenerative disc disease, wherein the patient has been treated by the method of  claim 11 . 
     
     
         13 . An intervertebral disc in vivo, said disc having been injected with injectable disc regeneration fluid according to  claim 2 , 
     
     
         14 . A method of treating a patient presenting with signs of hydrodynamic intervertebral disc dysfunction, the method comprising
 diagnosing hydrodynamic disc dysfunction in at least one intervertebral disc of said patient;   testing said at least one intervertebral disc of said patient to establish cellular proteoglycan production within said at least one disc; and   injecting a plurality of purified cell growth factors into said at least one disc to treat a patient presenting with signs of hydrodynamic intervertebral disc dysfunction.   
     
     
         15 . The method of  claim 14  wherein at least two of said plurality of cell growth factors are bone-derived. 
     
     
         16 . An injectable cell growth medium for intervertebral disc regeneration, said medium comprising
 an injectable fluid comprising digestion-resistant remodelable collagen, said collagen being cross-linked through photooxidative catalysis and irradiation by visible light; and   a plurality of purified cell growth factors dispersed within said fluid to form an injectable cell growth medium.   
     
     
         17 . The injectable cell growth, medium of  claim 16  wherein at least two of said plurality of cell growth factors are bone-derived. 
     
     
         18 . An injectable disc regeneration fluid for intervertebral discs, the material comprising
 injectable cell growth medium according to  claim 17 ; and   cells responsive to said injectable cell growth medium through proteoglycan elaboration in vivo.   
     
     
         19 . The disc regeneration fluid of  claim 18  wherein said cells are chondrocytes. 
     
     
         20 . The disc regeneration fluid of  claim 18  wherein said cells are mesenchymal stem cells. 
     
     
         21 . The disc regeneration fluid of  claim 20  wherein said cells are human-derived. 
     
     
         22 . The disc regeneration fluid of  claim 20  wherein said cells are cultured in vitro to increase their response to said cell growth factors. 
     
     
         23 . An injectable material for treating a patient for hydrodynamic disc dysfunction, the material made by a process comprising
 cross-linking collagen through photooxidative catalysis and irradiation by visible light;   purifying a plurality of bone-derived cell growth factors;   dispersing said purified bone-derived cell growth factors within said cross-linked collagen; and   dispersing cells responsive to said purified plurality of bone-derived cell growth factors within said cross-linked collagen to form an injectable material for treating hydrodynamic disc dysfunction.   
     
     
         24 . The injectable material of  claim 23  wherein said cells are chondrocytes. 
     
     
         25 . The injectable material of  claim 23  wherein said cells are mesenchymal stem cells. 
     
     
         26 . The injectable material of  claim 24  wherein said cells are human-derived. 
     
     
         27 . The injectable material of  claim 24  wherein said cells are cultured in vitro to increase their response to said cell growth factors. 
     
     
         28 . A method of hydrating an intervertebral disc annulus fibrosus in vivo, the method comprising
 testing said disc for cellular proteoglycan production within said disc; and   injecting cell growth medium according to  claim 16  into said disc to hydrate the annulus fibrosus.   
     
     
         29 . A method of reducing susceptibility to herniation of an intervertebral disc in a patient having a history of intervertebral disc herniation, the method comprising
 testing said disc for cellular proteoglycan production within said disc; and   injecting cell growth medium according to  claim 16  into said disc to reduce susceptibility to herniation through hydration of the annulus fibrosus.   
     
     
         30 . A method of increasing the height of a patient presenting with hydrodynamic disc dysfunction in at least one intervertebral disc, the method comprising
 testing said at least one disc for cellular proteoglycan production within said at least one disc; and   injecting cell growth medium according to  claim 16  into said at least one disc to increase the height of said patient by increasing intervertebral spacing through increased proteoglycan production in said at least one disc.   
     
     
         31 . An injectable cell suspension for treating a patient having degenerative disc disease, the suspension comprising
 an injectable fluid comprising a plurality of purified cell growth factors; and   a plurality of living cells dispersed within said injectable fluid to form an injectable cell suspension for treating degenerative disc disease, said cells being responsive to said cell growth factors by increased elaboration of proteoglycans.   
     
     
         32 . The injectable cell suspension of  claim 31  wherein said cells are chondrocytes. 
     
     
         33 . The injectable cell suspension of  claim 31  wherein said cells are mesenchymal stem cells. 
     
     
         34 . The injectable cell suspension of  claim 33  wherein said cells are human-derived. 
     
     
         35 . The injectable cell suspension of  claim 31  wherein said cells are cultured in vitro to increase their response to said cell growth factors. 
     
     
         36 . The injectable cell suspension of  claim 31  wherein at least two of said plurality of cell growth factors are bone-derived. 
     
     
         37 . A method of treating a patient presenting with degenerative disc disease, the method comprising
 providing an injectable cell growth medium according to  claim 16 ; and   injecting said injectable cell growth medium into at least one of said patient's intervertebral discs to treat degenerative disc disease in said disc.   
     
     
         38 . A method of treating a patient presenting with hydrodynamic disc dysfunction, the method comprising
 providing an injectable material according to  claim 23 ; and   injecting said injectable material into at least one of said patient's intervertebral discs to treat hydrodynamic disc dysfunction in said disc.   
     
     
         39 . A method of treating a patient presenting with degenerative disc disease, the method comprising
 providing an injectable cell suspension according to  claim 31 ; and   injecting said injectable cell suspension into at least one of said patient's intervertebral discs to treat degenerative disc disease in said disc.   
     
     
         40 . A method of cross-linking collagen to make digestion-resistant remodelable cross-linked collagen, the method comprising
 providing a hydrogel comprising collagen;   containing said hydrogel within a semipermeable membrane, said membrane being substantially transparent to visible light and substantially permeable to at least one photooxidative catalyst;   transporting at least one photooxidative catalyst through said semipermeable membrane and into said hydrogel; and   irradiating said hydrogel with visible light to cross-link said collagen.   
     
     
         41 . The method of  claim 40  comprising an additional step between said containing step and said transporting step, the additional step being
 submerging said hydrogel-containing semipermeable membrane in a high salt:high sucrose solution for about 24 to about 72 hours. 
 
     
     
         42 . The method of  claim 41  wherein said hydrogel comprises nucleus pulposus tissue. 
     
     
         43 . The method of  claim 42  wherein said transporting step occurs at a substantially constant hydrogel temperature of about 10° C. 
     
     
         44 . Digestion-resistant remodelable cross-linked collagen made by the method of  claim 43 . 
     
     
         45 . The method of  claim 41  wherein said hydrogel comprises substantially Type II collagen. 
     
     
         46 . The method of  claim 43  wherein said semipermeable membrane comprises dialysis tubing having a molecular weight cutoff of about 3500 Daltons. 
     
     
         47 . The method of  claim 43  wherein said transporting step and said irradiating step are substantially simultaneous. 
     
     
         48 . The method of  claim 43  wherein said transporting step occurs while said semipermeable membrane containing said hydrogel is immersed in a solution comprising at least one photooxidative catalyst. 
     
     
         49 . The method of  claim 43  comprising an additional step after said irradiating step, the additional step being
 terminating said transporting step and said irradiating step when said collagen cross-linking is substantially complete. 
 
     
     
         50 . The method of  claim 49  comprising an additional step after said terminating step, the additional step being
 extracting said cross-linked collagen from said hydrogel. 
 
     
     
         51 . The method of  claim 43  wherein said at least one photooxidative catalyst comprises methylene blue. 
     
     
         52 . A fluid matrix for treating intervertebral disc disease in a vertebrate, said fluid comprising nucleus pulposus tissue of a donor vertebrate. 
     
     
         53 . The fluid matrix of  claim 52  wherein said nucleus pulposus tissue is cross-linked. 
     
     
         54 . The fluid matrix of  claim 53  further comprising a growth factor. 
     
     
         55 . The fluid matrix of  claim 54  further comprising a plurality of living cells. 
     
     
         56 . The fluid matrix of  claim 55 , wherein said plurality of living cells comprise chondrocytes. 
     
     
         57 . The fluid matrix of  claim 55 , wherein said plurality of living cells comprise mesenchymal stem cells. 
     
     
         58 . The fluid matrix of  claim 55 , wherein said plurality of living cells are human-derived. 
     
     
         59 . The fluid matrix of  claim 52 , wherein said nucleus pulposus tissues are decellularized.