Platelet-Rich Fibrin Increases BMP2 Expression in Oral Fibroblasts via Activation of TGF-β Signaling.

Zahra Kargarpour,Layla Panahipour,Goran Mitulović,Reinhard Gruber,Jila Nasirzade,Richard J. Miron

doi:10.3390/ijms22157935

Zahra Kargarpour, Layla Panahipour + Show 4 more

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https://doi.org/10.3390/ijms22157935

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Abstract

Solid platelet-rich fibrin (PRF), consisting of coagulated plasma from fractionated blood, has been proposed to be a suitable carrier for recombinant bone morphogenetic protein 2 (BMP2) to target mesenchymal cells during bone regeneration. However, whether solid PRF can increase the expression of BMPs in mesenchymal cells remains unknown. Proteomics analysis confirmed the presence of TGF-β1 but not BMP2 in PRF lysates. According to the existing knowledge of recombinant TGF-β1, we hypothesized that PRF can increase BMP2 expression in mesenchymal cells. To test this hypothesis, we blocked TGF-β receptor 1 kinase with SB431542 in gingival fibroblasts exposed to PRF lysates. RT-PCR and immunoassays confirmed that solid PRF lysates caused a robust SB431542-dependent increase in BMP2 expression in gingival fibroblasts. Additionally, fractions of liquid PRF, namely platelet-poor plasma (PPP) and the buffy coat (BC) layer, but not heat-denatured PPP (Alb-gel), greatly induced the expression of BMP2 in gingival fibroblasts. Even though PRF has no detectable BMPs, PRF lysates similar to recombinant TGF-β1 had the capacity to provoke canonical BMP signaling, as indicated by the nuclear translocation of Smad1/5 and the increase in its phosphorylation. Taken together, our data suggest that PRF can activate TGF-β receptor 1 kinase and consequently induce the production of BMP2 in cells of the mesenchymal lineage.

Highlights

Platelet-rich fibrin (PRF) is generated by centrifuging blood, aiming to remove erythrocytes and to obtain a fraction consisting of plasma enriched with platelets and leucocytes
We recently performed a proteomic analysis of PRF lysates showing the presence of the classical growth factor TGF-β1, but neither of the BMP family members was identified [18]
We examined our data from another independent proteomic analysis of the total PRF clots based on mass spectrometric analysis

Summary

Introduction

Platelet-rich fibrin (PRF) is generated by centrifuging blood, aiming to remove erythrocytes and to obtain a fraction consisting of plasma enriched with platelets and leucocytes. PRF membranes are applied clinically to defects with the overall goal of supporting the natural processes of wound healing and bone regeneration [4,5], which require the coordinated action of growth factors, including those released from solid PRF membranes [6,7]. Apart from its intrinsic growth factors, solid PRF can serve as a carrier for the delivery of recombinant growth factors [8]. Among these proposed growth factors is bone morphogenetic protein (BMP2), a member of the transforming growth factor beta (TGF-β) superfamily that is characterized by its osteoinductive potential [8,9]. PRF might serve as a carrier for transplanted cells producing BMP2 [14]. The question of whether PRF can stimulate the expression of BMP2 in potential target cells remains

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