Concentrated growth factor exudate enhances the proliferation of human periodontal ligament cells in the presence of TNF-α

The purpose of this study was to evaluate the effects of concentrated growth factor exudate (CGFe) on human periodontal ligament cells (hPDLCs) stimulated by tumor necrosis factor (TNF)-α. From the peripheral blood of healthy donors, CGFe was prepared according to the Sacco protocol after 7 days of incubation. The hPDLCs were cultured by a tissue explant method and identified with anti-vimentin and anti-cytokeratin antibodies. Cells were subjected to four different treatments: i) Control; ii) TNF-α (10 ng/ml); iii) CGFe (concentration 50%); and iv) CGFe+TNF-α. The proliferation of hPDLCs was measured with Cell Counting Kit-8 assays. Osteogenic differentiation and mineralization were determined by Alizarin Red S staining, alkaline phosphatase activity, western blotting and reverse transcription-quantitative polymerase chain reaction. CGFe enhanced cell proliferation and upregulated ALP activity, the mineralization level, and osteogenic-associated osteocalcin, runt-related transcription factor 2 and Osterix gene expression in hPDLCs under inflammatory conditions induced by TNF-α. The present study demonstrated that CGFe enhanced hPDLC proliferation and osteogenic differentiation in the presence of TNF-α-induced inflammation. As CGFe can be obtained from the venous blood of patients, it generates no immune reaction. Thus, it is more economical and beneficial to use CGFe in clinical periodontal regeneration practice than synthetic growth factors.