Electrospun nanofibers of poly(ε-caprolactone)/depolymerized chitosan for respiratory tissue engineering applications

Synthetic grafts comprised of a porous scaffold in the size and shape of the natural tracheobronchial tree, and autologous stem cells have shown promise in the ability to restore the structure and function of a severely damaged airway system. For this specific application, the selected scaffold material should be biocompatible, elicit limited cytotoxicity, and exhibit sufficient mechanical properties. In this research, we developed composite nanofibers of polycaprolactone (PCL) and depolymerized chitosan using the electrospinning technique and assessed the properties of the fibers for its potential use as a scaffold for regenerating tracheal tissue. Water-soluble depolymerized chitosan solution was first prepared and mixed with polycaprolactone solution making it suitable for electrospinning. Morphology and chemical structure analysis were performed to confirm the structure and composition of the fibers. Mechanical testing of nanofibers demonstrated both elastic and ductile properties depending on the ratio of PCL to chitosan. To assess biological potential, porcine tracheobronchial epithelial (PTBE) cells were seeded on the nanofibers with composition ratios of PCL/chitosan: 100/0, 90/10, 80/20, and 70/30. Transwell inserts were modified with the nanofiber membrane and cells were seeded according to air-liquid interface culture techniques that mimics the conditions found in the human airways. Lactase dehydrogenase assay was carried out at different time points to determine cytotoxicity levels within PTBE cell cultures on nanofibers. This study shows that PCL/chitosan nanofiber has sufficient structural integrity and serves as a potential candidate for tracheobronchial tissue engineering.