A novel approach for hyaluronic acid (HA) crosslinking via diglycidyl ether (DGE) was investigated for scaffolds fabrication. In particular, HA sponges were obtained by lyophilization and then reacted with 1,4-butandiol diglycidyl ether (BDDGE) in heterogeneous conditions. Insoluble matrices with 4-20% of the reactive sites of HA modified were produced. The hydrogels showed high swelling capability depending on external stimuli; when equilibrated in physiological solution, pore ranged from 70 to130μm and G' values were in the range 2-10kPa. The matrices proved highly stable in cell culture conditions and to enzymatic degradation. A biological evaluation revealed good cellular viability within the scaffolds in two weeks experiments. The main achievement consists in that the novel conditions (BDDGE and heterogeneous reaction) permitted to obtain insoluble, directly structured and potentially applicable scaffolds with the lower content of crosslinker reported to date. The in vitro characterization outcomes propose the hydrogels as promising substrates for soft tissues regeneration.