Fabrication of bilayer electrospun polycaprolactone-collagen-chitosan nanofibers for corneal tissue engineering
Fatemeh Basiri1 , Mohammad Ali Derakhshan2 *, Fatemeh Sanie-Jahromi3
- Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran
- . Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran
- Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract: Cornea, as the outermost segment of the eye, plays significant roles in refracting light to the retina and keeping the eye from various damages and infections due to its high transparency and strength. Also, the majority of oxygen and nutrition for the anterior segment of the eyes are provided by this part. Various physical and chemical damages and diseases such as trauma, burn and keratoconus may impair the normal function of cornea. Transplantation of cornea is conventionally considered to replace the damaged cornea, however, lack of enough donors and immune rejections are yet the main limitations. Therefore, designing a scaffold that mimics the main structure of native cornea is demanded
Methods: Electrospinning was utilized to fabricate double-layered structures out of polycaprolactone, collagen and chitosan. Random nanofibers of PCL-Col were prepared as the outer layer to support corneal epithelial cells and aligned PCL-Col-Cs nanofibers were fabricated as the inner layer to mimic corneal stroma. Morphology and diameter of the nanofibers and also, double-layer structure was investigated by SEM. To analyze the chemical composition of the nanofibers, ATR-FTIR was performed. Water contact angle analyses and water absorption capacity of the nanofibrous structures were evaluated. Also, tensile properties of the nanofibers was investigated. Keratocyte were isolated from donated human corneal stroma and were cultured on the prepared nanofibers for cytocompatibility/toxicity evaluations by alamar blue assay
Results: Findings of characterization of the nanofibers by SEM illustrated proper formation of smooth and bead-less nanofibers and continuous shaping of the double-layered structure. ATR-FTIR spectra showed the bonds and functional groups in the composition of PCL, collagen and chitosan. Also, the results proved the appropriate mechanical properties of the nanofibrous structures. Cell study in 5 days illustrated the biocompatibility of the prepared nanofibers. Also, SEM image showed the attachment and spreading of the cell on the scaffolds, exhibiting the elongation of the cells according to the orientation of the aligned fibers.
Conclusion: Designing a bilayer electrospun nanofibers out of polycaprolactone, collagen and chitosan would be considered as a potential biocompatible construct for corneal tissue engineering to support keratocyte functions.
