Integrated proteomics highlights functional activation induced by advanced-platelet rich fibrin plus (A-PRF +) in primary equine fibroblasts

: Wounds are common in equine practice, and often lead to complications such as infections, delayed healing and hypertrophic scarring, which can be costly and difficult to manage. Developing affordable and effective treatments has become an increasingly important focus in veterinary research. Equine advanced-platelet-rich fibrin plus (A-PRF+) demonstrates regenerative properties comparable to its human counterpart, but cellular-level investigations exploring its molecular mechanisms remain limited. This study aimed to investigate the in vitro effects of equine A-PRF + on primary fibroblast cell cultures. The secretome analysis of A-PRF + revealed a complex protein profile involved in matrix remodelling, cell proliferation, and inflammation. Treatment with this platelet concentrate resulted in increased cell proliferation, enhanced migration, and significant changes in cell cycle progression compared to control groups. Reactive oxygen species production and organelles metabolism stimulation were observed, indicating active cellular responses, as well as an increase in genes and proteins associated with cell proliferation and wound regeneration. Proteomic analysis of treated fibroblasts confirmed the differential expression of key proteins associated with extracellular matrix dynamics and tissue regeneration processes. These findings provide insights into the molecular profile and functional responses of equine fibroblasts exposed to A-PRF + , contributing to our understanding of its cellular effects, supporting further exploration of this product in regenerative medicine applications.