Platelet-rich fibrin stimulates the proliferation and expression of proteins related to survival, adhesion, and angiogenesis in gingival fibroblasts cultured on a titanium nano-hydroxyapatite-treated surface.

Objectives: This in vitro study aimed to evaluate the cell viability and expression of proteins related to angiogenesis, adhesion, and cell survival (vascular endothelial growth factor, paxillin, vinculin, fibronectin, and protein kinase B) in gingival fibroblasts that were cultured on titanium discs treated with or without nanohydroxyapatite and exposed to platelet-rich fibrin (PRF)-conditioned medium.

Methods: To obtain the conditioned medium, the PRF membranes were prepared and incubated for 48 h in a culture medium without fetal bovine serum. Analyses were performed at 24 and 48 h for the cells cultured on machined-titanium discs or surfaces treated with nanohydroxyapatite in a control medium or PRF-conditioned medium, resulting in four experimental groups (CT-TI, CT-NANO, PRF-TI, and PRF-NANO).

Results: A decrease in the viability of the gingival fibroblasts was not observed in any of the experimental groups. The PRF-NANO group showed significantly higher immunoexpression of paxillin and AKT at 24 and 48 h (p < 0.01). The same result was observed for vinculin expression at 24 h (p < 0.001). The expression of fibronectin at 48 h and VEGF at 24 and 48 h was significantly higher when the cells were exposed to the PRF-conditioned medium, regardless of the disc surface (p < 0.05).

Conclusion: Gingival fibroblasts cultured on a nanohydroxyapatite-treated surface and in a PRF-conditioned medium showed a greater expression of proteins modulating adhesion, angiogenesis, and cell survival. Our results may contribute to the understanding of the mechanisms related to peri-implant soft tissue sealing.