Compositions for regeneration and repair of cartilage lesions

Abstract

Disclosed is a cartilage repair product that induces both cell ingrowth into a bioresorbable material and cell differentiation into cartilage tissue. Such a product is useful for regenerating and/or repairing both vascular and avascular cartilage lesions, particularly articular cartilage lesions, and even more particularly mensical tissue lesions, including tears as well as segmental defects. Also disclosed is a method of regenerating and repairing cartilage lesions using such a product.

1. A product for repair of cartilage lesions, comprising:
 a. a cartilage repair matrix suitable for conforming to a defect in cartilage; and  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: transforming growth factor β1 (TGFβ1), bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7; 
 wherein the quantity of said TGFβ1 in said mixture is greater than 1% of total proteins in said mixture;  
 wherein the quantity of said BMP-2 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 wherein the quantity of said BMP-3 in said mixture is from about 0.1% to about 15% of total proteins in said mixture; and,  
 wherein the quantity of said BMP-7 in said mixture is from about 0.01% to about 10% of total proteins in said mixture.  
 
 
     
     
       2. The product of  claim 1 , wherein the quantity of said TGFβ1 in said mixture is at least about 10% of total proteins in said mixture. 
     
     
       3. The product of  claim 1 , wherein said mixture of proteins further comprises TGFβ2, TGFβ3, BMP-4, BMP-5, BMP-6, cartilage-derived morphogenetic protein (CDMP), FGF-I, osteocalcin, osteonectin, BSP, lysyloxidase, cathepsin L pre, albumin, transferrin, Apo Al LP and Factor XIIIb. 
     
     
       4. A product for repair of cartilage lesions, comprising:
 a. a cartilage repair matrix; and  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: 
 (i) a bone-derived osteogenic or chondrogenic formulation containing at least one bone morphogenetic protein (BMP); and,  
 (ii) a TGFβ protein that is exogenous to said formulation of (i);  
 
 wherein the weight ratio of said exogenous TGFβ protein to total BMP in said mixture of proteins is greater than about  10:1; and,  
 wherein said exogenous TGFβ protein is present in an amount sufficient to increase cartilage induction by said composition over a level of cartilage induction by said bone-derived osteogenic or chondrogenic protein formulation in the absence of said exogenous TGFβ protein.  
 
     
     
       5. The product of  claim 4 , wherein said exogenous TGFβ protein is TGFβ1. 
     
     
       6. The product of any one of  claim 1  or  5 , wherein the weight ratio of TGFβ1 to all other proteins in said mixture of proteins is at least about  1:10. 
     
     
       7. The product of any one of  claim 1  or  5 , wherein the weight ratio of TGFβ1 to all other proteins in said mixture of proteins is at least about  10:1. 
     
     
       8. The product of  claim 4 , wherein said mixture of proteins comprises TGFβ superfamily proteins consisting of: TGFβ1, bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7, wherein said TGFβ superfamily proteins comprise from about 0.5% to about 99.99% of said mixture of proteins. 
     
     
       9. The product of  claim 4 , wherein said bone-derived osteogenic or chondrogenic formulation comprises TGFβ1, TGFβ2, TGFβ3, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, CDMP, FGF-I, osteocalcin, osteonectin, BSP, lysyloxidase, cathepsin L pre, albumin, transferrin, Apo Al LP and Factor XIIIb. 
     
     
       10. A product for repair of cartilage lesions, comprising:
 a. a cartilage repair matrix; and  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: 
 (i) a TGFβ protein; and,  
 (ii) at least one bone morphogenetic protein (BMP);  
 
 wherein the weight ratio of said TGFβ protein to total BMP in said mixture of proteins is greater than about  10:1.  
 
     
     
       11. The product of any one of  claim 1 ,  4  or  10 , wherein said mixture of proteins comprises Bone Protein (BP). 
     
     
       12. The product of any one of  claim 1 ,  4  or  10 , wherein said cartilage-inducing composition is at a concentration of from about 0.5% to about 33% by weight of said product. 
     
     
       13. The product of any one of  claim 1 ,  4  or  10 , wherein said cartilage repair matrix is bioresorbable. 
     
     
       14. The product of  claim 1 ,  4  or  10 , wherein said cartilage repair matrix comprises collagen from bovine tendon. 
     
     
       15. The product of  claim 10 , wherein said mixture of proteins comprises TGFβ superfamily proteins consisting of: TGFβ1, bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7, wherein said TGFβ superfamily proteins comprise from about 0.5% to about 99.99% of said mixture of proteins. 
     
     
       16. The product of any one of  claim 8  or  15 , wherein said TGFβ superfamily proteins comprise from about 0.5% to about 25% of said mixture of proteins. 
     
     
       17. The product of any one of  claim 8  or  15 , wherein the quantity of said TGFβ1 in said mixture is from about 0.01% to about 75% of total proteins in said mixture. 
     
     
       18. The product of any one of  claim 1 ,  8  or  15 , wherein the quantity of said TGFβ1 in said mixture is from about 33% to about 99.99% of total proteins in said mixture. 
     
     
       19. The product of any one of  claim 1 ,  8  or  15 , wherein said mixture of proteins further comprises at least one bone matrix protein selected from the group consisting of osteocalcin, osteonectin, bone sialoprotein (BSP), lysyloxidase, cathepsin L pre, osteopontin, matrix GLA protein (MGP), biglycan, decorin, proteoglycan-chondroitin sulfate III (PG-CS III), bone acidic glycoprotein (BAG-75), thrombospondin (TSP) and fibronectin; wherein said bone matrix protein comprises from about 20% to about 98% of said mixture of proteins. 
     
     
       20. The product of any one of  claim 1 ,  8  or  15 , wherein said mixture of proteins further comprises at least one growth factor protein selected from the group consisting of fibroblast growth factor-I (FGF-I), FGF-II, FGF-9, leukocyte inhibitory factor (LIF), insulin, insulin-like growth factor I (IGF-I), IGF-II, platelet-derived growth factor AA (PDGF-AA), PDGF-BB, PDGF-AB, stromal derived factor-2 (SDF-2), pituitary thyroid hormone (PTH), growth hormone, hepatocyte growth factor (HGF), epithelial growth factor (EGF), transforming growth factor-α (TGFα) and hedgehog proteins; wherein said growth factor protein comprises from about 0.01% to about 50% of said mixture of proteins. 
     
     
       21. The product of any one of  claim 1 ,  8  or  15 , wherein said composition further comprises one or more serum proteins. 
     
     
       22. The product of any one of  claim 4  or  10 , wherein the weight ratio of said TGFβ protein to total BMP in said mixture of proteins is greater than about 100:1. 
     
     
       23. A product for repair of cartilage lesions, comprising:
 a. a cartilage repair matrix suitable for conforming to a defect in cartilage; and  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: transforming growth factor β1 (TGFβ1), bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7; 
 wherein the quantity of said TGFβ1 in said mixture is greater than 1% of total proteins in said mixture;  
 wherein the quantity of said BMP-2 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 wherein the quantity of said BMP-3 in said mixture is at least about 0.38% of total proteins in said mixture; and,  
 wherein the quantity of said BMP-7 in said mixture is from about 0.01% to about 10% of total proteins in said mixture.  
 
 
     
     
       24. The product of  claim 10 , wherein said BMP protein is selected from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9 and cartilage-derived morphogenetic protein (CDMP). 
     
     
       25. The product of  claim 10 , wherein said mixture of proteins comprises TGFβ1, TGFβ2, TGFβ3, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, cartilage-derived morphogenetic protein (CDMP), FGF-I, osteocalcin, osteonectin, BSP, lysyloxidase, cathepsin L pre, albumin, transferrin, Apo Al LP and Factor XIIIb. 
     
     
       26. A method for repair of cartilage lesions, comprising implanting and fixing into a cartilage lesion a product comprising:
 a. a cartilage repair matrix suitable for conforming to a defect in cartilage; and  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: transforming growth factor β1 (TGFβ1), bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7; 
 wherein the quantity of said TGFβ1 in said mixture is greater than 1% of total proteins in said mixture;  
 wherein the quantity of said BMP-2 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 wherein the quantity of said BMP-3 in said mixture is from about 0.1% to about 15% of total proteins in said mixture; and,  
 wherein the quantity of said BMP-7 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 
 
       whereby implanting and fixing said product into said cartilage lesion enhances repair of said defect in cartilage as compared to in the absence of said product. 
     
     
       27. A method for repair of cartilage lesions, comprising implanting and fixing into a cartilage lesion a product comprising:
 a. a cartilage repair matrix; and,  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: 
 (i) a bone-derived osteogenic or chondrogenic formulation of proteins containing at least one bone morphogenetic protein (BMP); and,  
 (ii) a TGFβ protein that is exogenous to said formulation of (i);  
 
 wherein the weight ratio of said exogenous TGFβ protein to total BMP in said mixture of proteins is greater than about  10:1; and,  
 wherein said exogenous TGFβ protein is present in an amount sufficient to increase cartilage induction by said composition over a level of cartilage induction by said bone-derived osteogenic or chondrogenic protein formulation in the absence of said exogenous TGFβ protein;  
 
       whereby implanting and fixing said product into said cartilage lesion enhances repair of said defect in cartilage as compared to in the absence of said product. 
     
     
       28. The method of  claim 27 , wherein said TGFβ protein is TGFβ1. 
     
     
       29. The method of any one of  claim 26  or  28 , wherein the weight ratio of TGFβ1 to all other proteins in said mixture of proteins is at least about 1:10. 
     
     
       30. The method of any one of  claim 26  or  28 , wherein the weight ratio of TGFβ1 to all other proteins in said mixture of proteins is at least about 10:1. 
     
     
       31. A method for repair of cartilage lesions, comprising implanting and fixing into a cartilage lesion a product comprising:
 a. a cartilage repair matrix; and,  
 b. a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: 
 (i) a TGFβ protein; and,  
 (ii) at least one bone morphogenetic protein (BMP);  
 
 wherein the weight ratio of said TGFβ protein to total BMP in said mixture of proteins is greater than about  10:1;  
 
       whereby implanting and fixing said product into said cartilage lesion enhances repair of said defect in cartilage as compared to in the absence of said product. 
     
     
       32. The method of  claim 31 , wherein said TGFβ protein is TGFβ1. 
     
     
       33. The method of any one of  claim 26 ,  27  or  31 , wherein said cartilage lesion is an articular cartilage lesion. 
     
     
       34. The method of any one of  claim 26 ,  27  or  31 , wherein said cartilage lesion is a meniscal cartilage lesion. 
     
     
       35. The method of  claim 34 , wherein said lesion is a vascular meniscus lesion. 
     
     
       36. The method of  claim 34 , wherein said lesion is an avascular meniscus lesion. 
     
     
       37. The method of any one of  claim 27  or  31 , wherein the weight ratio of said TGFβ protein to total BMP in said mixture of proteins is greater than about 100:1. 
     
     
       38. The product of  claim 10 , wherein said TGFβ protein is TGFβ1. 
     
     
       39. A method for repair of segmental cartilage lesions, comprising implanting and fixing into a segmental cartilage lesion:
 a. a first product comprising: 
 (i) a cartilage repair matrix configured as a sheet; and  
 (ii) a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: transforming growth factor β1 (TGFβ1), bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7; 
 wherein the quantity of said TGFβ1 in said mixture is greater than 1% of total proteins in said mixture;  
 wherein the quantity of said BMP-2 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 wherein the quantity of said BMP-3 in said mixture is from about 0.1% to about 15% of total proteins in said mixture; and,  
 wherein the quantity of said BMP-7 in said mixture is from about 0.01% to about 10% of total proteins in said mixture; and,  
 
 
 b. a second product comprising a cartilage repair matrix configured to replace cartilage removed from a segmental lesion;  
 
       wherein said second product is implanted into said lesion and wherein said first product is implanted between an edge of said lesion and said second product to provide an interface between said lesion and said second product. 
     
     
       40. The method of  claim 39 , wherein said second product further comprises a cartilage-inducing composition contained on or within said matrix comprising a mixture of proteins comprising: transforming growth factor β1 (TGFβ1), bone morphogenetic protein (BMP)-2, BMP-3, and BMP-7;
 wherein the quantity of said TGFβ1 in said mixture is greater than 1% of total proteins in said mixture;  
 wherein the quantity of said BMP-2 in said mixture is from about 0.01% to about 10% of total proteins in said mixture;  
 wherein the quantity of said BMP-3 in said mixture is from about 0.1% to about 15% of total proteins in said mixture; and,  
 wherein the quantity of said BMP-7 in said mixture is from about 0.01% to about 10% of total proteins in said mixture.  
 
     
     
       41. The method of any one of  claim 26 ,  27  or  31 , wherein said lesion is a tear and wherein said matrix is configured as a sheet, wherein said step of implanting comprises inserting said product directly into said tear. 
     
     
       42. A composition comprising:
   at least one TGFβ protein; and        at least one bone morphogenetic protein  ( BMP );      wherein the weight ratio of said at least one TGFβ protein to said at least one BMP is a chondrogenesis inducing ratio greater than  10 : 1 .     
     
     
       43. The composition of  claim 42 , wherein the at least one TGFβ protein is TGFβ 1 , TGFβ 2 , TGFβ 3 , TGFβ 4 , TGFβ 5 , or a mixture thereof. 
     
     
       44. The composition of  claim 43 , wherein the at least one TGFβ protein is TGFβ 1 . 
     
     
       45. The composition of  claim 42 , wherein the at least one TGFβ protein is exogenous. 
     
     
       46. The composition of  claim 42 , wherein the weight ratio of the at least one TGFβ protein to the at least one bone morphogenetic protein is greater than about  100 : 1 . 
     
     
       47. The composition of  claim 46 , wherein the weight ratio of the at least one TGFβ protein to the at least one bone morphogenetic protein is greater than about  1 , 000 : 1 . 
     
     
       48. The composition of  claim 47 , wherein the weight ratio of the at least one TGFβ protein to the at least one bone morphogenetic protein is greater than about  10 , 000 : 1 . 
     
     
       49. The composition of  claim 42 , wherein the at least one bone morphogenetic protein is BMP-   2 , BMP -   3 , BMP -   4 , BMP -   5 , BMP -   6 , BMP -   7 , BMP -   8 , BMP -   9 , CDMP, or a mixture thereof.   
     
     
       50. The composition of  claim 49 , wherein the at least one bone morphogenetic protein is BMP-   2 , BMP -   3 , BMP -   7 , or a mixture of two or more thereof.   
     
     
       51. The composition of  claim 49 , wherein the at least one bone morphogenetic protein is BMP-   2 .   
     
     
       52. The composition of  claim 49 , wherein the at least one bone morphogenetic protein is BMP-   3 .   
     
     
       53. The composition of  claim 49 , wherein the at least one bone morphogenetic protein is BMP-   7 .   
     
     
       54. The composition of  claim 49 , wherein the at least one bone morphogenetic protein comprises BMP-   2 , BMP -   3 , and BMP -   7 .   
     
     
       55. The composition of  claim 54 , wherein the at least one TGFβ protein is TGFβ 1 . 
     
     
       56. The composition of  claim 42 , wherein the at least one TGFβ protein or the at least one bone morphogenetic protein are constituents of a bone- derived protein mixture.   
     
     
       57. The composition of  claim 56 , wherein the at least one TGFβ protein and the at least one bone morphogenetic protein are constituents of a bone- derived protein mixture.   
     
     
       58. The composition of  claim 57 , wherein the protein mixture is Bone Protein ( BP ). 
     
     
       59. The composition of  claim 56 , wherein the protein mixture comprises the TGFβ superfamily proteins TGFβ 1 , BMP-   2 , BMP -   3 , and BMP -   7 , and wherein said TGFβ superfamily proteins comprise from about  0 . 5   %  to about  99 . 99   %  of said protein mixture.   
     
     
       60. The composition of  claim 59 , wherein said TGFβ superfamily proteins comprise from about  0 . 5 %  to about  25   %  of said protein mixture.   
     
     
       61. The composition of  claim 59 , wherein the quantity of said TGFβ 1  in said mixture is from about  0 . 01 %  to about  75   %  of total proteins in said mixture.   
     
     
       62. The composition of  claim 56 , wherein the TGFβ protein is exogenous to the protein mixture. 
     
     
       63. The composition of  claim 62 , wherein said exogenous TGFβ protein is TGFβ 1 . 
     
     
       64. The composition of  claim 63 , wherein the quantity of TGFβ 1  is from about  0 . 1 %  to about  1   %  of total proteins in the mixture.   
     
     
       65. The composition of  claim 56 , wherein the protein mixture comprises a quantity of TGFβ 1  that is at least about  10 %  of the total proteins in said protein mixture.   
     
     
       66. The composition of  claim 65 , wherein the weight ratio of TGFβ 1  to all other proteins in said protein mixture is at least about  10 : 1 . 
     
     
       67. The composition of  claim 56 , wherein the mixture of proteins comprises TGFβ superfamily proteins including TGFβ-   1 , BMP -   2 , BMP -   3  and BMP -   7  and wherein the TGFβ superfamily proteins comprise from about  0 . 5   %  to about  99 . 99   %  of the mixture of proteins.   
     
     
       68. The composition of  claim 67 , wherein the quantity of TGFβ 1  in the protein mixture is from about  33 %  to about  99 . 99   %  of total proteins in said mixture.   
     
     
       69. The composition of  claim 56 , wherein said mixture of proteins further comprises at least one bone matrix protein selected from the group consisting of osteocalcin, osteonectin, bone sialoprotein ( BSP ) , lysyloxidase, cathepsin L pre, osteopontin, matrix GLA protein  ( MGP ) , biglycan, decorin, proteoglycan - chondroitin sulfate III  ( PG - CS III ) , bone acidic glycoprotein  ( BAG -   75   ) , thrombospondin  ( TSP )  and fibronectin; wherein said bone matrix protein comprises from about  20   %  to about  98   %  of said mixture of proteins.   
     
     
       70. The composition of  claim 56 , wherein said mixture of proteins further comprises at least one growth factor protein selected from the group consisting of fibroblast growth factor- I  ( FGF - I ) , FGF - II, FGF -   9 , leukocyte inhibitory factor  ( LIF ) , insulin, insulin - like growth factor I  ( IGF - I ) , IGF - II, platelet - derived growth factor AA  ( PDGF - AA ) , PDGF - BB, PDGF - AB, stromal derived factor -   2   ( SDF -   2   ) , pituitary thyroid hormone  ( PTH ) , growth hormone, hepatocyte growth factor  ( HGF ) , epithelial growth factor  ( EGF ) , transforming growth factor -α ( TGF α)  and hedgehog proteins; wherein said growth factor protein comprises from about  0 . 01   %  to about  50   %  of said mixture of proteins.   
     
     
       71. The composition of  claim 56 , wherein said composition further comprises one or more serum proteins. 
     
     
       72. The composition of  claim 42 , further comprising a matrix including collagen. 
     
     
       73. The composition of  claim 42 , further comprising a matrix including a gel. 
     
     
       74. The composition of  claim 42 , further comprising a matrix including a polymer. 
     
     
       75. The composition of  claim 42 , further comprising an injectable matrix. 
     
     
       76. The composition of  claim 42 , further comprising a matrix having an acidic pH. 
     
     
       77. The composition of  claim 42 , wherein the composition is adapted to repair cartilage lesions selected from articular lesions, vascular meniscal lesions, and avascular meniscal lesions. 
     
     
       78. The composition of  claim 77 , wherein the composition is adapted to repair articular cartilage lesions. 
     
     
       79. The composition of  claim 42 , further comprising a matrix configured as a sheet. 
     
     
       80. The composition of  claim 42 , further comprising a bioresorbale matrix and wherein the at least one TGFβ protein and the at least one bone morphogenetic protein are about  0 . 5 %  to about  33   %  by weight of the composition.