Thiolated hyaluronan-based hydrogels cross-linked using oxidized glutathione

Abstract

The invention provides methods, compositions and kits relating to hyaluronan based matrices using oxidized glutathione as a crosslinking agent.

1 .- 7 . (canceled) 
     
     
         8 . A method of making the hydrogel of  claim 40 , the method comprising: (a) contacting thiolated carboxymethylated hyaluronan (CMHA-S) with GSSG; and (b) adding thiolated gelatin and a therapeutic agent to the CMHA-S and GSSG of step (a), thereby forming a hydrogel comprising the therapeutic agent, CMHA-S and thiolated gelatin. 
     
     
         9 . A method of making the hydrogel of  claim 31 , the method comprising contacting a polymer comprising a thiol group with the maleimido-tagged peptide to form a peptide functionalized polymer; and contacting the peptide functionalized polymer with GSSG to form a peptide functionalized hydrogel. 
     
     
         10 .- 27 . (canceled) 
     
     
         28 . A hydrogel comprising a first thiolated polymer and a second thiolated polymer crosslinked with GSSG. 
     
     
         29 . The hydrogel of  claim 28 , wherein the first thiolated polymer is CMHA-S. 
     
     
         30 . The hydrogel of  claim 29 , wherein the second thiolated polymer is thiolated gelatin. 
     
     
         31 . The hydrogel of  claim 28 , wherein the first thiolated polymer comprises a maleimido-tagged peptide. 
     
     
         32 . The hydrogel of  claim 31 , wherein the peptide comprises RGD. 
     
     
         33 . The hydrogel of  claim 28 , wherein the first thiolated polymer, the second thiolated polymer, or the first thiolated polymer and the second thiolated polymer comprise different monomer units. 
     
     
         34 . The hydrogel of  claim 28 , which has a shear elastic modulus ranging from about 9 to about 37 Pa. 
     
     
         35 . The hydrogel of  claim 29 , which comprises between about 0.1% to about 5% w/v CMHA-S. 
     
     
         36 . The hydrogel of  claim 35 , which comprises between about 0.1 mM to about 54 mM GSSG. 
     
     
         37 . The hydrogel of  claim 36 , which comprises about 0.8% CMHA-S and about 4 mM GSSG. 
     
     
         38 . The hydrogel of  claim 28 , which comprises between about 0.1 mM to about 54 mM GSSG. 
     
     
         39 . The hydrogel of  claim 28 , wherein the first thiolated polymer is thiolated CMHA-S and the second thiolated polymer is thiolated gelatin, and wherein the thiolated polymers are at a ratio of between about 1:1 to about 5:1 thiolated CMHA-S to thiolated gelatin. 
     
     
         40 . The hydrogel of  claim 28 , further comprising a therapeutic agent. 
     
     
         41 . The hydrogel of  claim 40 , wherein the therapeutic agent is a cell, a small molecule, a biologic, a peptide, a protein, a nucleic acid, or any combination thereof. 
     
     
         42 . The hydrogel of  claim 41 , wherein the cell is a cell of the central nervous system, a cell of the digestive system, a hepatocyte, a pancreatic cell, a retinal pigmented epithelial cell, an adipose cell, or any combination thereof. 
     
     
         43 . The hydrogel of  claim 40 , wherein the therapeutic agent is an antibiotic, an anti-viral, an anti-cancer drug, a growth factor, a hormone, a cytokine, an anti-inflammatory drug, a lipid, a polysaccharide, a nervous system modulator, a pain reliever, a narcotic, an antibody or any combination thereof. 
     
     
         44 . The hydrogel of  claim 40 , wherein the therapeutic agent binds to one or more components of the hydrogel through one or more covalent bond, ionic bond, hydrogen bond, van der Waals forces. 
     
     
         45 . The hydrogel of  claim 28 , which is formulated for subcutaneously administration or administration to the subconjunctival of a subject.