Development of chitosan membrane using non-toxic crosslinkers for potential wound dressing applications

There is a myriad of ways to crosslink hydrogel wound dressings; however, they require additional steps to remove the residue of the crosslinking agents, or their byproducts in biological environments are toxic. In this study, we studied and characterized the crosslinking of the chitosan hydrogels by various dicarboxylic acids, including oxalic acid, adipic acid, and sebacic acid under vacuum at 90 °C. The concentrations of the crosslinkers in the crosslinked hydrogels are tolerable for the cells, and the membranes can be used after crosslinking without complicated additional steps to remove the unreacted residues. The molar ratio of the crosslinkers was calculated based on the stoichiometry of the chitosan amine groups. Attenuated total reflectance Fourier transform infrared spectroscopy revealed amide linkage formation between amine groups of the chitosan and carboxyl groups of the dicarboxylic acids at 90 °C. The results showed that the chitosan membranes crosslinked with oxalic acid had higher Young's modulus (~ 1042 N/mm²) and ultimate tensile strength (~ 75 N/mm²) in comparison with the other dicarboxylic acids. Moreover, the membranes crosslinked with oxalic acid showed a weight loss of ~ 5.4% after 24 h at double-distilled water, which was drastically lower than that of the others. Thus, oxalic acid was selected as the most effective crosslinker. Cell viability assay, using mouse fibroblast (L929) cells, was conducted on the mechanically optimized membranes. The fibroblast cells successfully attached and spread well on the surface of the membranes. In conclusion, the obtained results suggested oxalic acid as an effective and non-toxic crosslinker for chitosan-based membranes for wound dressing applications.