Chemically Modified Gellan Gum Hydrogels with Tunable Properties for Use as Tissue Engineering Scaffolds

Abstract

<p>Gellan gum is a naturally occurring polymer that can cross-link in the presence of divalent cations to form biocompatible hydrogels. However, physically cross-linked gellan gum hydrogels lose their stability under physiological conditions, thus restricting the applications of these hydrogels in vivo. To improve the mechanical strength of the gels, we incorporated methacrylate into the gellan gum and chemically cross-linked the hydrogel through three polymerization methods: step growth through thiol–ene photoclick chemistry, chain-growth via photopolymerization, and mixed model in which both mechanisms were employed. Methacrylation was confirmed and quantified by proton nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy. The mechanical properties and chemistry of the cross-linked gels were systematically altered by varying the reaction conditions. The compression moduli of the resulting hydrogels ranged between 6.4 and 17.2 kPa. The swelling ratios of the hydrogels were correlated with the compression moduli and affected by the addition of calcium. In vitro enzymatic degradation rate was found to depend on the degree of methacrylation. NIH/3T3 fibroblast cell proliferation and morphology were related to substrate stiffness, with a high stiffness leading generally to higher proliferation. The proliferation is further affected by the thiol–ene ratio. These results suggest that a hydrogel platform based on the gellan gum can offer versatile chemical modifications and tunable mechanical properties. The influence of these substrates on cell behavior suggests that the gellan gum hydrogels have the flexibility to be engineered for a variety of biomaterials applications.</p>