Platelet-Rich Plasma Prevents In Vitro Transforming Growth Factor-β1-Induced Fibroblast to Myofibroblast Transition: Involvement of Vascular Endothelial Growth Factor (VEGF)-A/VEGF Receptor-1-Mediated Signaling

• Published in: Cells (Basel, Switzerland) Switzerland), 2018-09, Vol.7 (9), p.142
• Main Authors: Chellini, Flaminia, Tani, Alessia, Vallone, Larissa, Nosi, Daniele, Pavan, Paola, Bambi, Franco, Zecchi Orlandini, Sandra, Sassoli, Chiara
• Format: Article
• Language: English
• Subjects: Actin; Antibodies; Biotechnology; Blocking antibodies; Blood platelets; Collagen; confocal immunofluorescence; Connexin 43; Experiments; Fibroblasts; Fibrosis; Fluorescence microscopy; Kinases; Medicine; Molecular modelling; myofibroblasts; Notch-1; Notch1 protein; Phenotypes; Plasma; platelet-rich plasma (PRP); Platelets; Polymerase chain reaction; Smad3 protein; Smooth muscle; transforming growth factor (TGF)-β1/Smad3; Transforming growth factor-b1; Vascular endothelial growth factor; vascular endothelial growth factor (VEGF)-A; Vascular endothelial growth factor receptors; VEGFR-1/flt-1; Vinculin; Western blotting; α-smooth muscle actin
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells7090142

The antifibrotic potential of platelet-rich plasma (PRP) is controversial. This study examined the effects of PRP on in vitro transforming growth factor (TGF)-β1-induced differentiation of fibroblasts into myofibroblasts, the main drivers of fibrosis, and the involvement of vascular endothelial growth factor (VEGF)-A in mediating PRP-induced responses. The impact of PRP alone on fibroblast differentiation was also assessed. Myofibroblastic phenotype was evaluated by confocal fluorescence microscopy and western blotting analyses of α-smooth muscle actin (sma) and type-1 collagen expression, vinculin-rich focal adhesion clustering, and stress fiber assembly. Notch-1, connexin 43, and VEGF-A expression were also analyzed by RT-PCR. PRP negatively regulated fibroblast-myofibroblast transition via VEGF-A/VEGF receptor (VEGFR)-1-mediated inhibition of TGF-β1/Smad3 signaling. Indeed TGF-β1/PRP co-treated fibroblasts showed a robust attenuation of the myofibroblastic phenotype concomitant with a decrease of Smad3 expression levels. The VEGFR-1 inhibition by KRN633 or blocking antibodies, or VEGF-A neutralization in these cells prevented the PRP-promoted effects. Moreover PRP abrogated the TGF-β1-induced reduction of VEGF-A and VEGFR-1 cell expression. The role of VEGF-A signaling in counteracting myofibroblast generation was confirmed by cell treatment with soluble VEGF-A. PRP as single treatment did not induce fibroblast myodifferentiation. This study provides new insights into cellular and molecular mechanisms underpinning PRP antifibrotic action.