Abstract
BACKGROUND/PURPOSE: Extracellular matrix (ECM) may be useful as a natural scaffold for tissue engineering. Since dental follicle stem cells (DFSCs) share a similar embryonic origin with corneal keratocytes and endothelial cells and possess the ability to proliferate and differentiate, in this study, we investigate the potential of DFSCs to differentiate into corneal cells under the supercritical carbon dioxide (SC-CO(2)) decellularized porcine corneal ECM culture stimulation. MATERIALS AND METHODS: DFSCs were seeded onto the SC-CO(2) decellularized porcine corneal ECM and the surrounding culture dish for 21 days. Cell morphology, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and immunofluorescence staining were used to evaluate the potential of DFSCs to differentiate into corneal epithelial, keratocyte, or endothelial cells. The culture medium was collected every 3 days for growth factor analysis. RESULTS: The DFSCs cultured on the ECM exhibit notable cell-ECM adhesion and the potential to primarily differentiate into keratocytes, with a partial capacity for differentiation into corneal endothelial cells, while the cells outside the ECM are weakly affected. Growth factor array analysis revealed changes in growth factor expression in different culture conditions, DFSCs, ECM, and DFSCs/ECM co-culture at various time points, indicating a dynamic shift in the microenvironment. CONCLUSION: The present study demonstrated that DFSCs cultured on the porcine corneal ECM can differentiate mainly into keratocytes-like and partially into corneal endothelial-like cells, exhibiting strong cell-ECM adhesion and regulation of growth factors. Provide insights into the therapeutic application potential of dental-derived stem cells and SC-CO(2) decellularized tissue for corneal diseases.