Glycosaminoglycan-Based Materials as an Engineered Biocompatible Cellular Matrix

Abstract

Disclosed herein is a cross-linked polymeric system comprising thiolated hyaluronic acid (HA), thiolated chondroitin sulfate (CS), and functionalized polyethylene glycol (PEG), wherein said functionalized PEG cross-links thiolated HA and thiolated CS. Methods of fabrication and utilization of the same are also claimed. This polymeric system may be used as an engineered biocompatible cellular matrix for 3D cell culture, tissue engineering and regenerative medicine applications.

1 . A composition comprising thiolated hyaluronic acid (HA), thiolated chondroitin sulfate (CS) and a functionalized polyethylene glycol (PEG) derivative, wherein said PEG derivative crosslinks thiolated HA and thiolated CS. 
     
     
         2 . The composition of  claim 1  wherein said functionalized PEG derivative contains a plurality of activated vinyl groups. 
     
     
         3 . The composition of  claim 1  wherein the molecular ratio of the total thiol group of HA and CS vs. said activated vinyl group is maintained constantly as about 1.07. 
     
     
         4 . The composition of  claim 1  wherein said activated vinyl groups are selected from the group consisting of poly (ethylene glycol) diacrylate (PEGDA), poly (ethylene glycol) Divinyl Sulfone (PEGVS), and 4-arm poly (ethylene glycol) vinyl Sulfone (4PEGVS). 
     
     
         5 . The composition of  claim 1  wherein thiolated HA further comprising a plurality of amino groups (—NH 2 ) groups selected from the following formula I-IV: 
       
         
           
           
               
               
           
         
         wherein the functional groups R 1 , R 2 , R 3 , R 4 , and R 5  comprise any one of or a combination of haloacetates, dihydrazides, amines, thiols, carboxylic acids, aldehydes, ketones, active hydrogen sites on aromatic ring, dienes, azide isothiocyanates, isocyanates, acyl azides, NHS esters, sulfo-NHS, sulfonyl chloride, epoxides, carbonates, aryl halides, imidoesters, carbodiimides (e.g. DCC and EDC), alkylphosphate compounds, anhydrides, fluorophenyl esters, hydroxymethyl phosphines, guanidino groups, iodoacetyl derivatives, maleimides, aziridines, acryloyl derivatives, arylating agents, disulfide derivatives, vinylsulfone, phenylthioester, cisplatins, diazoacetates, carbonyl diimidazoles, oxiranes, N, N′-disuccinimidyl carbonates, N-hydroxylsuccinimidyl chloroformates, alkyl halogens, hydrazines, alkynes, and phosphorus-bound chlorine. 
       
     
     
         6 . An osteochondral regenerative engineering composite comprising poly (lactide-co-glycolide) (PLGA) grafted to a composition via —NH 2  group to form a bone mimetic, wherein said composition comprising thiolated hyaluronic acid (HA), thiolated chondroitin sulfate (CS) and a functionalized polyethylene glycol (PEG) derivative, wherein said PEG derivative crosslinks thiolated HA and thiolated CS. 
     
     
         7 . The osteochondral regenerative engineering composite of  claim 6  wherein said PLGA is made of lactic acid/glycolic acid at a ratio of about 85:15. 
     
     
         8 . The composition of  claim 1  wherein the molecular weight and length of said PEG derivatives are adjustable to modify the composite modular storage and loss value. 
     
     
         9 . The composition of  claim 8 , wherein the molecular weight of said PEG derivatives ranges from about 700 Da to about 8000 Da. 
     
     
         10 . A method of making a composition of cross-linked HA, CS and PEG comprising the steps of: preparing thiolated HA;
 a. preparing thiolated CS;   b. preparing PEG derivative containing a plurality of activated functionalities;   c. mixing said HA, CS, and PEG derivative in an aqueous medium; and   d. initiating cross-linking;   wherein said activated functionality is elected from the group consisting of alkoxysulfonate, arylsulfonate, heteroarylsulfonate, maleimido, ether NHS esters, sulfo-NHS,   wherein the structures of said PEG derivative is any one of or a combination of linear, dendrimers-like, star-shaped, hyper-branched, combed, brushed, cross-linked architectures, fibers, microspheres, and nanoparticles.   
     
     
         11 . The method of  claim 10 , wherein the activated functionality of PEG comprises any one of or a combination of isothiourea, isourea, amide, sulfonamide, secondary amine, sulfonamide, shift-base, secondary amino-methyl, carbamate, aryl amine, amidine, amide, phosphoramidate, guanidine, substituted imidocarbonate, thioether, 4-amino derivative of cytosine, aryl thioether, disulfide, sulfonate, β-thiosulfonyl, ester, carbamate, hydrazone, diazo, triazoles, iodinated compound, carbohydrates, amino acid esters bond, cycloalkene, oxime triazole, and triazoline. 
     
     
         12 . The composition of  claim 1  further comprising functionalized peptides selected from the group consisting of arginine-glycine-aspartate (RGD), fibronectin, laminin, and fibrinogen, wherein said peptides are functionalized by carboxyl, amine or thiol group, and conjugated with HA, CS, or functionalized PEG derivatives through thiol-ene click reactions and esterification. 
     
     
         13 . The composition of  claim 1  further comprising tissue engineering cells, wherein the cells are selected from the group consisting of mesenchymal stem cells, osteoblast, chondrocytes, adipocyte, fibroblast, hepatocytes, enterocytes, urothelial cells, blood cells, skin cells, endothelial cells, nerve cells, sex cells, cancer cells and combination thereof. 
     
     
         14 . The composition of  claim 1  further comprising small molecules as therapeutic agents, 
     
     
         15 . The composition of  claim 1  further comprising at least one growth factor.