Injectable polysaccharide hydrogels as localized nitric oxide delivery formulations

A series of injectable polysaccharide hydrogels were prepared with oxidized dextran and diethylenetriamine-modified carboxymethylcellulose or hyaluronic acid. Rheological evaluation revealed that carboxymethylcellulose-based hydrogels achieved the largest storage moduli (>1 kPa) when prepared from 5 wt. % solutions. However, carboxymethylcellulose-based hydrogels with storage moduli >100 Pa were prepared from solutions with concentrations as low as 2 wt. %. Hyaluronic acid-based hydrogels demonstrated smaller storage moduli but had swelling ratios more than four times that of the carboxymethylcellulose systems at the same polymer concentrations. The incorporation of N-diazeniumdiolate NO donors into the hydrogels resulted in reduced hydrogel storage moduli as a function of NO donor concentration. The impact of the hydrogel architecture on NO-release kinetics proved dependent on the identity of the NO donor. Hydrogel degradation over 14 d was measured at pH 5.4 and 7.4 and indicated that hyaluronic acid-based hydrogels degraded more rapidly than carboxymethylcellulose hydrogels and that the addition of NO to the hydrogels increased the rate at which they degraded. In vitro cytotoxicity of hydrogel extracts was evaluated against five cell lines, with no observed toxicity except for that of hyaluronic acid-based hydrogel extracts against human gingival fibroblasts. The diverse properties, versatility, and non-toxic characteristics of these injectable hydrogels should facilitate local delivery of nitric oxide for a range of biomedical applications.