Abstract
BACKGROUND: Platelet concentrates have been used in tissue regeneration. The purpose of this study was to examine effects of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF) and concentrated growth factor (CGF) on the osteogenic differentiation of periodontal ligament fibroblasts (PDLFs). METHODS: Leukocyte- and platelet-rich fibrins, CGFs and PDLFs were obtained from New Zealand rabbits. The release of basic fibroblast growth factor (bFGF), bone morphogenetic protein 2 (BMP-2) and transforming growth factor β1 (TGF-β1) from L-PRFs and CGFs was measured at 5 h and 1, 3, 5, 7 days, using the enzyme linked immunosorbent assay. The PDLFs were treated with exudates of L-PRF or CGF. After the treatment, cell counting kit-8 assay was performed at day 1, 3, 5 and 7. Alkaline phosphatase (ALP) assay and Western blotting were applied at day 7. Three blocking antibodies were used to neutralize the proteins of bFGF, BMP-2 and TGF-β1. RESULTS: Leukocyte- and platelet-rich fibrin and CGF showed different growth factor release pattern, but similar accumulated concentration of these growth factors. PDLFs proliferation was significantly promoted by both L-PRF and CGF at day 1, 3 and 7, and CGF group was superior to L-PRF group at day 1 and 3. Both L-PRF and CGF significantly enhanced PDLFs ALP activity and protein expression of osteogenic markers. The osteopontin level was higher in CGF group than in L-PRF group, but no significant differences were found between two groups for ALP activity. Three blocking antibodies significantly downregulated both L-PRF and CGF induced osteogenic markers expression. CONCLUSION: Both CGF and L-PRF can promote the proliferation and osteogenic differentiation of PDLFs. The bFGF, BMP-2 and TGF-β1 are involved in both L-PRF and CGF induced osteogenic differentiation of PDLFs.