Effect ofadvanced platelet-rich fibrin + (A-PRF+) modified by gold nanoparticles on osteoblastic differentiation of human mesenchymal stem cells- in vitro

Abstract

Background: Among the many types of platelet concentrates, platelet-rich fibrin (PRF) is widely used to accelerate regeneration of different issues. Despite the therapeutic effects of PRF in soft tissue restoration, there is still insufficient clinical evidence of PRF beneficial effects on bone regeneration. Gold nanoparticles have been shown to enhance osteogenic differentiation and bone mineralization. The purpose of this study was to investigate the effect of A-PRF+ modified by gold nanoparticles on the osteoblastic differentiation of human mesenchymal stem cells. Methods: In this study, 8 blood samples were taken at a volume of 8 ml from four healthy individuals. After PRF preparation, gold nanoparticles were added to yield final concentration of 0.0125 mM. We analyzed the effect of A-PRF and gold nanoparticles on viability of stem cells by using MTT over a period of 7 days. We also measured osteogenic potential of A-PRF modified by gold nanoparticles conditioned media on stem cells by measuring alkaline phosphatase activity and existence of calcium deposition by Alizarin Red S staining. Results: MTT assay revealed 0.0125 mM gold nanoparticles had no cytotoxic effects on stem cells after 7 days. Addition of 0.0125 mM gold nanoparticle to A-PRF+ clot increased cell survival rate compared to the non-treated control group (p<0.05). 7-day incubation of stem cells with A-PRF+ modified by gold nanoparticles conditioned media was shown to promote alkaline phosphatase activity compared to the control cells and group treated with A-PRF+ condition media (p<0.05). By using Alizarin Red S staining, red-colored calcium deposits were observed in the group treated with A-PRF+ and gold nanoparticles conditioned media in comparison with non-treated cells (p<0.05). We also found that A-PRF+ conditioned medium was unable to predispose calcium deposition compared to the combination of A-PRF+ and gold nanoparticles (p<0.05). Conclusion: Adding gold nanoparticles to A-PRF+ and fibrin and platelets byproducts could be as an alternative strategy to improve osteogenic capacity of human mesenchymal stem cells.