Methods and compositions for regenerating connective tissue

Abstract

Connective tissue regenerative compositions and methods of repairing and regenerating connective tissue using such compositions are provided. The compositions generally comprise a bioactive hydrogel matrix comprising a polypeptide, such as gelatin, and a long chain carbohydrate, such as dextran. The hydrogel matrix may further include polar amino acids, as well as additional beneficial additives. Advantageously, the compositions include further components, such as osteoinductive or osteoconductive materials, medicaments, stem or progenitor cells, and three-dimensional structural frameworks. The compositions are useful for regenerating connective tissue, and can be administered to an area having injury to, or a loss of, connective tissue, such as bone, cartilage, tendon, and ligament.

1 . A method for regenerating connective tissue selected from bone, cartilage, ligament, and tendon, comprising administering a bioactive hydrogel matrix to a site in need of connective tissue regeneration, said bioactive hydrogel matrix comprising:
 a polypeptide;   a long chain carbohydrate;   one or more components selected from the group consisting of polar amino acids, polar amino acid analogs or derivatives, divalent cation chelators, and combination thereof; and   at least one osteoinductive or osteoconductive material selected from the group consisting of hydroxyapatite, demineralized bone matrix (DBM), bone morphogenetic proteins (BMPs), transforming growth factors (TGFs), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), platelet-derived growth factors (PDGFs), epidermal growth factors (EGFs), vascular endothelial growth factors (VEGFs), and vascular permeability factors (VPFs).   
     
     
         2 . The method of  claim 1 , wherein the polypeptide is a tissue-derived polypeptide derived from tissue selected from the group consisting of collagens, gelatins, keratin, decorin, aggrecan, and glycoproteins or is a tissue-derived polypeptide derived from extracts of tissue selected from the group consisting of submucosal tissues, arteries, vocal chords, pleura, trachea, bronchi, pulmonary alveolar septa, ligaments, auricular cartilage, abdominal fascia, liver, kidney, neurilemma, arachnoid, dura mater, and pia mater. 
     
     
         3 . The method of  claim 1 , wherein the polypeptide has a molecular mass of about 3,000 to about 3,000,000 Da. 
     
     
         4 . The method of  claim 1 , wherein the long chain carbohydrate is a polysaccharide or a sulfated polysaccharide. 
     
     
         5 . The method of  claim 4 , wherein the long chain carbohydrate is a polysaccharide selected from the group consisting of dextran, dextrin, heparan, heparin, hyaluronic acid, chondroitin, alginate, agarose, carageenan, amylopectin, amylose, glycogen, starch, cellulose, chitin, and chitosan, or the long chain carbohydrate is a sulfated polysaccharide selected from the group consisting of heparan sulfate, heparin sulfate chondroitin sulfate, dextran sulfate, dermatan sulfate, and keratan sulfate. 
     
     
         6 . The method of  claim 1 , wherein the long chain carbohydrate has a molecular mass of about 2,000 to about 8,000,000 Da. 
     
     
         7 . The method of  claim 1 , wherein the hydrogel matrix comprises one or more polar amino acids selected from the group consisting of tyrosine, cysteine, serine, threonine, asparagine, glutamine, aspartic acid, glutamic acid, arginine, lysine, histidine, and mixtures thereof. 
     
     
         8 . The method of  claim 1 , wherein the hydrogel matrix comprises ethylenediaminetetraacetic acid or a salt thereof. 
     
     
         9 . The method of  claim 1 , wherein the bioactive hydrogel matrix further comprises at least one medicament selected from the group consisting of antivirals, antibacterials, anti-inflammatories, immunosuppressants, analgesics, anticoagulants, and healing promotion agents. 
     
     
         10 . The method of  claim 1 , wherein the bioactive hydrogel matrix further comprises cells selected from the group consisting of stem cells, progenitor cells, and mixtures thereof. 
     
     
         11 . The method of  claim 1 , wherein, after said administering step, the bioactive hydrogel matrix is flowable and is at least partially contained within a three-dimensional structural framework. 
     
     
         12 . The method of  claim 11 , wherein the three-dimensional structural framework includes a bioreabsorbable material. 
     
     
         13 . The method of  claim 11 , wherein the three-dimensional framework includes a material selected from the group consisting of coralline, metals, sponge, bioactive glass, ceramics, calcium salts, collagen, keratin, fibrinogen, alginate, chitosan, allogenic bone, autologous bone, hyaluronan, polyethylene, poly(vinylidene fluoride), poly(tetrafluoroethylene), poly(vinyl alcohol), poly(hydroxyalkanoate), poly(ethylene terephthalate), poly(butylene terephthalate), poly(methy  1  methacrylate), poly(hydroxyethyl methacrylate), poly(N-isopropylacrylamide), poly(dimethyl siloxane), polydioxanone, and polypyrrole, poly(glycolic acid), poly(lactic acids), poly(ethylene oxides), poly(lactide-co-glycolides), poly(s-caprolactone), polyanhydrides, polyphosphazenes, poly(ortha-esters), and polyimides, and combinations thereof. 
     
     
         14 . The method of  claim 11 , wherein the three-dimensional structural framework comprises a metal cage or a collagen sponge. 
     
     
         15 . The method of  claim 1 , wherein the bioactive hydrogel matrix is in dehydrated form prior to said administering step. 
     
     
         16 . The method of  claim 15 , wherein the dehydrated bioactive hydrogel matrix is in particulate form. 
     
     
         17 . The method of  claim 16 , wherein the at least one osteoinductive or osteoconductive material is dispersed within the bioactive hydrogel matrix. 
     
     
         18 . The method of  claim 1 , wherein at least a portion of the bioactive hydrogel matrix is in crosslinked form, the long chain carbohydrate being covalently crosslinked to the polypeptide. 
     
     
         19 . The method of  claim 1 , wherein the osteoinductive or osteoconductive material comprises hydroxyapatite. 
     
     
         20 . The method of  claim 19 , wherein the osteoinductive or osteoconductive material comprises demineralized bone matrix (DBM). 
     
     
         21 . The method of  claim 1 , wherein the at least one osteoinductive or osteoconductive material is present at a concentration of about 0.01 volume percent to about 90 volume percent, based upon the total volume of the hydrogel matrix. 
     
     
         22 . The method of  claim 21 , wherein the at least one osteoinductive or osteoconductive material is present at a concentration of about 50 volume percent to about 80 volume percent, based upon the total volume of the hydrogel matrix. 
     
     
         23 . The method of  claim 1 , comprising administering the composition at a site undergoing surgical attachment or reattachment of the connective tissue. 
     
     
         24 . The method of  claim 1 , comprising administering the composition at a site of a torn anterior cruciate ligament (ACL), a rotator cuff injury, or damaged cartilage in a joint. 
     
     
         25 . The method of  claim 1 , wherein the polypeptide comprises gelatin. 
     
     
         26 . The method of  claim 25 , wherein the long chain carbohydrate comprises dextran. 
     
     
         27 . The method of  claim 26 , wherein bioactive hydrogel matrix further comprises at least one polar amino acid or divalent cation chelator. 
     
     
         28 . A method of treating a degenerative disease of the natural joint of a patient comprising administering a bioactive hydrogel matrix to a joint of a patient suffering from the degenerative disease, the bioactive hydrogel matrix comprising a polypeptide, a long chain carbohydrate, and at least one osteoinductive or osteoconductive material selected from the group consisting of hydroxyapatite, demineralized bone matrix (DBM), bone morphogenetic proteins (BMPs), transforming growth factors (TGFs), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), platelet-derived growth factors (PDGFs), epidermal growth factors (EGFs), vascular endothelial growth factors (VEGFs), and vascular permeability factors (VPFs). 
     
     
         29 . The method of  claim 28 , wherein the polypeptide comprises gelatin. 
     
     
         30 . The method of  claim 28 , wherein the bioactive hydrogel matrix further comprises at least one polar amino acid or divalent cation chelator. 
     
     
         31 . The method of  claim 28 , wherein the degenerative disease is arthritis. 
     
     
         32 . The method of  claim 31 , wherein the arthritis is osteoarthritis. 
     
     
         33 . The method of  claim 31 , comprising administering the bioactive hydrogel matrix prior to complete loss of cartilage in the joint. 
     
     
         34 . The method of  claim 28 , wherein said administering comprises injecting the bioactive hydrogel matrix into the joint. 
     
     
         35 . The method of  claim 28 , wherein, prior to said administering step, the bioactive hydrogel matrix is in a dehydrated form and wherein the method further comprises rehydrating the dehydrated bioactive hydrogel matrix. 
     
     
         36 . A method of promoting integration of an artificial joint into the surrounding tissue comprising administering a bioactive hydrogel matrix at the site of the artificial joint, the bioactive hydrogel matrix comprising a polypeptide, a long chain carbohydrate, and at least one osteoinductive or osteoconductive material selected from the group consisting of hydroxyapatite, demineralized bone matrix (DBM), bone morphogenetic proteins (BMPs), transforming growth factors (TGFs), fibroblast growth factors (FGFs), insulin-like growth factors (IGFs), platelet-derived growth factors (PDGFs), epidermal growth factors (EGFs), vascular endothelial growth factors (VEGFs), and vascular permeability factors (VPFs).