Cell Growth And Orientation Of Corneal Stroma Keratocytes On Electrospun Polycaprolactone-Collagen Scaffolds

Background & Aim Corneal cells, namely keratocytes, are a hallmark cell type for topical ocular drugs, eye drops, and drug toxicity studies. However, there has yet to exist in-vitro models of keratocytes mimicking natural corneal cells, which align in the direction of parallel collagen fibers in corneal stroma. By culturing cells on tissue-engineered scaffolds that closely mimic the structure of the stroma, cell growth and behavior should be improved. In this study, we investigated the effect of scaffold fiber orientation on the survivability and proliferation of corneal stroma keratocytes. Methods, Results & Conclusion Three different types of materials, polycaprolactone, polycaprolactone-jellyfish collagen blend, and polycaprolactone-bovine dermis atelocollagen blend, were electrospun to obtain nanofibrous scaffolds with both random and aligned fiber orientations. Keratocytes were cultured on the scaffolds over a duration of 4 days, and their survivability and cellular alignment was determined using a live and dead cell assay on day 1 and day 4. In addition, cell proliferation was measured using 4′,6-diamidino-2-phenylindole (DAPI) staining on day 1 and day 4. The results show that cells survive and proliferate better on the aligned fibers by 2-fold, with the orientation of cell growth matching that of the fibers, as shown in Fig. 1. Moreover, polycaprolactone-atelocollagen blended scaffolds favor cell survivability and proliferation as compared to pure polycaprolactone and polycaprolactone-jellyfish collagen scaffolds. Our polycaprolactone-atelocollagen scaffolds have potential applications in scaffold-based drug screening, drug toxicity testing and tissue-engineering for corneal regeneration.