Genipin cross-linked electrospun chitosan-based nanofibrous mat as tissue engineering scaffold

Objective(s): To improve water stability of electrospun chitosan/ Polyethylene oxide (PEO) nanofibers, genipin, a biocompatible and nontoxic agent, was used to crosslink chitosan based nanofibers.  Materials and Methods: Different amounts of genipin were added to the chitosan/PEO solutions, chitosan/PEO weight ratio 90/10 in 80 % acetic acid, and the solutions were then electrospun to form nanofibers. The spun nanofibers were exposed to water vapor to complete crosslinking. The nanofibrous membranes were subjected to detailed analysis by scanning electron microscopy (SEM), Fourier transform infrared-attenuated total reflection (FTIR-ATR) spectroscopy, swelling test, MTT cytotoxicity, and cell attachment.   Results: SEM images of electrospun mats showed that genipin-crosslinked nanofibers retained their fibrous structure after immerging in PBS (pH=7.4) for 24 hours, while the uncrosslinked samples lost their fibrous structure, indicating the water stability of genipin-crosslinked nanofibers. The genipin-crosslinked mats also showed no significant change in swelling ratio in comparison with uncrosslinked ones. FTIR-ATR spectrum of uncrosslinked and genipin-crosslinked chitosan nanofibers revealed the reaction between genipin and amino groups of chitosan. Cytotoxicity of genipin-crosslinked nanofibers was examined by MTT assay on human fibroblast cells in the presence of nanofibers extraction media. The genipin-crosslinked nanofibers did not show any toxic effects on fibroblast cells at the lowest and moderate amount of genipin. The fibroblast cells also showed a good adhesion on genipin-crosslinked nanofibers.   Conclusion: This electrospun matrix would be used for biomedical applications such as wound dressing and scaffold for tissue engineering without the concern of toxicity.