Platelet-Rich Plasma in a Murine Model: Leukocytes, Growth Factors, Flt-1, and Muscle Healing

• Published in: The American journal of sports medicine 2016-08, Vol.44 (8), p.1962-1971
• Main Authors: Denapoli, Priscila Martins Andrade, Stilhano, Roberta Sessa, Ingham, Sheila Jean McNeill, Han, Sang Won, Abdalla, Rene Jorge
• Format: Article
• Language: English
• Subjects: Animals; Blood platelets; Contusions - drug therapy; Contusions - metabolism; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Humans; Leukocytes - metabolism; Male; Mice; Mice, Inbred C57BL; Muscles - injuries; Muscles - metabolism; Muscles - physiopathology; Plasma; Platelet-Derived Growth Factor - metabolism; Platelet-Rich Plasma - chemistry; Sports medicine; Transforming Growth Factor beta1 - metabolism; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; Vascular Endothelial Growth Factor Receptor-1 - metabolism; Wound Healing - drug effects
• ISSN: 0363-5465; 1552-3365
• DOI: 10.1177/0363546516646100

Background: It is well known that platelet-rich plasma (PRP) preparations are not the same and that not all preparations include white blood cells, but the part that leukocytes play on the healing role of PRP is still unknown. Purpose: The primary aim of this study was to evaluate the influence of leukocytes in different PRP preparations with a special emphasis on growth factor concentrations. The secondary aim was to evaluate the influence of PRP on muscle healing. Study Design: Controlled laboratory study. Methods: Two PRP preparation procedures were evaluated. Blood fractions were stained with Rapid Panoptic, and growth factors (transforming growth factor beta 1 [TGF-β1], vascular endothelial growth factor [VEGF], insulin-like growth factor [IGF], epidermal growth factor [EGF], hepatocyte growth factor [HGF], and platelet-derived growth factor [PDGF]) were quantified by enzyme-linked immunosorbent assay. Western blotting analysis was performed for Fms-related tyrosine kinase 1 (Flt-1). A muscle contusion injury was created and treated with PRP at different time points. Results: Leukocytes were the main source of VEGF, and all other growth factors measured had a higher concentration in the preparations that included the buffy coat and consequently had a higher concentration of white blood cells. Flt-1 was also found in platelet-poor plasma (PPP). There were higher concentrations of PDGF and HGF in the preparations that encompassed the buffy coat. A PRP injection 7 days after the injury provided significantly increased exercise performance and decreased the fibrotic area when compared with other PRP-treated groups. Conclusion: VEGF is only present in PRP′s buffy coat, while Flt-1 is present in PPP. A PRP injection 7 days after an injury resulted in improved exercise performance. Clinical Relevance: The presence of Flt-1 in PRP provides yet another explanation for results described in the literature after a PRP injection. This information is relevant for selecting the best PRP for each type of injury.