Biological Effects of Advanced Platelet-Rich Fibrin+ (A-PRF+)-Conditioned Medium on Primary Equine Corneal Stromal Cells: A Proof-of-Concept Study

Source abstract

Corneal wound healing largely depends on the activation of stromal fibroblasts, which coordinate extracellular matrix remodeling and tissue regeneration. Although advanced platelet-rich fibrin (A-PRF+) has emerged as a promising biomaterial in regenerative medicine, its biological effects on equine corneal fibroblasts remain poorly understood. In this study, primary equine corneal fibroblasts were isolated from stromal explants and treated with an A-PRF+-conditioned medium. Cell metabolic activity, viability, intracellular reactive oxygen species, wound healing, cell cycle progression, growth factor secretion, and collagen type I expression were evaluated using colorimetric assays, fluorescence imaging, flow cytometry, and ELISA. A-PRF+ significantly enhanced fibroblast metabolic activity, increased intracellular ROS production, accelerated wound closure and cell cycle re-entry, and stimulated the secretion of PDGF-BB, TGF-β1, and VEGF-A. Furthermore, treatment increased the percentage of collagen type I-positive fibroblasts, supporting enhanced extracellular matrix production. Collectively, these findings demonstrate that A-PRF+ exerts beneficial effects on primary equine corneal fibroblasts by stimulating their metabolic, proliferative, migratory, and secretory activities. These results provide an experimental basis for the use of A-PRF+-based eye drops as a regenerative strategy to improve corneal healing in horses.