Preparation and characterization of genipin cross-linked porous chitosan–collagen–gelatin scaffolds using chitosan–CO2 solution

Novel porous scaffolds composed of chitosan, collagen and gelatin were prepared by the multistep procedure involving final freeze-drying and characterized. To eliminate the need for residual acid removal from the material after drying, carbon dioxide saturation process was used for chitosan blend formulation. The use of CO2 for chitosan dissolution made the scaffold preparation process more reproducible and economically sustainable. Genipin was applied to stabilize the structure of the scaffolds and those crosslinked at a level of 7.3% exhibited a homogenous porous structure (33.1%), high swelling capacity (27.6 g/g for wound exudate like medium; 62.5 g/g for water), and were stable under cyclic compression. The values of other investigated parameters: dissolution degree (30%), lysozyme-induced degradation (5% after 168 h), good antioxidant properties (DPPH, ABTS, Fe2+ assays) and especially very low in vitro cytotoxicity against fibroblasts (103%, MTT assay), were highly advantageous for possible biomedical applications of the novel materials.