Standardized Platelet-Rich Fibrin (PRF) from canine and feline origin: An analysis on its secretome pattern and architectural structure.

Platelet-rich fibrin (PRF) has been incorporated in surgical procedures to promote tissue and bone healing, particularly in human medicine. The rationale for the use of platelet-based products stems from the fact that platelets, after being activated, release growth factors (GFs) and other active molecules such as cytokines, that modulate inflammation and tissue repair. Although PRF has been advanced as a therapeutic treatment for veterinary use, namely in canine and feline patients (following human medicine developments), to our knowledge a full characterization of PRF therapeutic effectors has never been performed. Herein, we studied the biological properties and release profile of GFs and other cytokines throughout ten days in in vitro culture conditions, in order to investigate the potential therapeutic ability of PRF for canine and feline practice. A protocol for obtaining PRF from whole blood without anti-coagulant from both species was optimized, originating large and homogenous PRF clots. Then, PRF clots obtained from four dogs and four cats were incubated in culture medium to assess the temporal release of platelet-derived growth factor-BB (PDGF-BB), vascular endothelial factor-A (VEGF-A), transforming growth factor β-1 (TGF-β1), and interleukin-8 (IL-8). Furthermore, morphological characterization of PRF clots, fresh and after 10 days of incubation, was performed by histology and high-resolution field emission electron scanning microscopy. In standard culture conditions, PRF clots from both species released PDGF-BB, TGF- β1 and VEGF-A, in a sustained manner, up to day 10. Moreover, PRF presents an initial burst release of IL-8, a mediator of inflammatory response which plays a key role in neutrophil recruitment and degranulation. Overall, our findings show that PRF clots may be an efficient therapeutic strategy in canine and feline clinical practice, accelerating the local healing mechanism, through the sustained delivery of signalling molecules involved in the healing cascade.