Surface Modification of Eggshell Membrane with Electrospun Chitosan/Polycaprolactone Nanofibers for Enhanced Dermal Wound Healing

Eggshell membrane (ESM), a naturally occurring microfibrous biopolymer network comprising collagen I, V, and X, GAGs, and other significant proteins, is responsible for guided tissue regeneration. The extraction methodology of ESM and surface topography of the microfibers impede its extensive usage in skin tissue engineering. Herein we deploy a unique route of ESM surface modification utilizing chitosan/poly-caprolactone (CS/PCL) nanofibers to fabricate a bilayered scaffold for wound healing application. Microstructural and surface topographic analysis of the construct confirms the bilayered structure of the composite with smooth nanofibers of CS/PCL decorated on ESM. The two layers were cross-linked by carbodiimide chemistry as confirmed by XPS and FTIR analysis. Cytocompatibility of the scaffolds was evaluated with human dermal fibroblast (HDF) cells culture study. The biomimetic architecture and composition of modified ESM facilitated extensive cell adhesion, migration, and proliferation while an impeded cell adhesion was observed on the natural tissue. Moreover, owing to the presence of ESM, the scaffolds adhered naturally to the wound bed while implanted on a full-thickness wound in a rat model. Further, the nanofiber modified ESM group showed extensive host cell migration and proliferation thus leading to faster re-epithelization and dermal regeneration with high collagen deposition in comparison to natural ESM. The above in vitro and in vivo results substantiate the effect of nanofiber functionalization on the ESM surface thus making the bilayered construct a potential dermal substitute.