Degradable thiol-acrylate hydrogels as tunable matrices for three-dimensional hepatic culture

A degradable poly(ethylene glycol)-diacrylate (PEGDA) hydrogel system was developed using simple macromer formulations and visible light initiated thiol-acrylate photopolymerization. In addition to PEGDA, other components in this gelation system include eosin-Y as a photosensitizer, bi-functional thiol (dithiothreitol, DTT) as a dual-purpose co-initiator and cross-linker, and N-vinylpyrrolidone (NVP) as a co-monomer. Gelation was achieved through a mixed-mode step-chain growth polymerization mechanism under bright visible light exposure. Increasing photosensitizer or NVP concentrations accelerated photo-crosslinking and increased final gel stiffness. Increasing bifunctional thiol content in the prepolymer solution only increased gel stiffness to some degree. As the concentration of thiol surpassed certain range, thiol-mediated chain-transfer events caused thiol-acrylate gels to form with lower degree of cross-linking. Pendant peptide, such as integrin ligand RGDS, was more effectively immobilized in the network via a thiol-acrylate reaction (using thiol-bearing peptide Ac-CRGDS. Underline indicates cross-linkable motif) than through homo-polymerization of acrylated peptide (e. g., acryl-RGDS). The incorporation of pendant peptide comes with the expense of a lower degree of gel cross-linking, which was rectified by increasing co-monomer NVP content. Without the use of any readily degradable macromer, these visible light initiated mixed-mode cross-linked hydrogels degraded hydrolytically due to the formation of thiol-ether-ester bonds following thiol-acrylate reactions. An exponential growth relationship was identified between the hydrolytic degradation rate and bifunctional thiol content in the prepolymer solution. Finally, we evaluated the cytocompatibility of these mixed-mode crosslinked degradable hydrogels using in situ encapsulation of hepatocellular carcinoma Huh7 cells. Encapsulated Huh7 cells remained alive and proliferated as time to form cell clusters. The addition of NVP at a higher concentration (0.3%) did not affect Huh7 cell viability but resulted in reduction of cell metabolic activity, which was accompanied by an elevated urea secretion from the encapsulated cells. (C) 2013 Wiley Periodicals, Inc.