Effects of Nano-Hydroxyapatite-Coated PRF on Gingiva-Derived Mesenchymal Stem Cells: In Vitro Study

Platelet-rich fibrin (PRF) has been widely used in regenerative dentistry because of its potential to support tissue regeneration. Recently, modifications in PRF preparation protocols and tube surface characteristics have attracted attention because of their possible influence on fibrin organization and biologic activity. The present in vitro study aimed to evaluate the effects of nano-hydroxyapatite platelet-rich fibrin (HA-PRF) on gingiva-derived mesenchymal stem cells (GMSCs) by comparing it with leukocyte platelet-rich fibrin (L-PRF) and titanium platelet-rich fibrin (T-PRF). Gingival tissue and venous blood samples were obtained from a systemically healthy male volunteer. PRF membranes were prepared using conventional glass tubes, nano-hydroxyapatite-coated tubes, and titanium tubes. GMSCs were isolated, characterized, and cultured with PRF membranes. Cell viability and metabolic activity were evaluated using MTT analysis. Apoptosis and necrosis rates were assessed by Annexin V/PI flow cytometry. VEGF and TGF-β1 release levels were determined by ELISA, whereas IL-1β, IL-6, and TNF-α gene expression levels were analyzed using qRT-PCR. The HA-PRF and L-PRF groups demonstrated higher cell viability values compared with the T-PRF group on day 7. Annexin V/PI analysis revealed no statistically significant differences between the groups in terms of apoptosis and necrosis. Growth factor release and cytokine gene expression profiles demonstrated time-dependent biologic responses in all PRF membranes. Within the limitations of this study, HA-PRF showed no evidence of cytotoxicity and demonstrated biologic responses comparable to those observed with conventional L-PRF. Both HA-PRF and L-PRF generally exhibited more favorable cellular responses than T-PRF under the present experimental conditions.