Mei-Jiao Yu1, Pi-Shan Yang, Shao-Hua Ge. 1. Department of Periodontology, School of Stomatology, Shandong University, Jinan 250012, China.
Abstract
OBJECTIVE: To study the biological effects of phenytoin (PHT) on cultured human periodontal ligament fibroblasts (hPDLF), and explore the possibility of its accelerating periodontal regeneration. METHODS: Increasing concentrations of PHT (1, 5, 20, 100, 500, 2 500 mg/L) were added into the medium of the fourth passage of cultured hPDLF, respectively. After co-incubated for 3 days, cell proliferation activity, the total amount of protein and alkaline phosphatase (ALP) activity were detected. Mineralized sodium and PHT (20, 100, 500 mg/L) were added into the medium of the fourth passage hPDLF. After co-incubated, the mineralized nodules formation were detected by Von Kossa staining. The third passage hPDLF were stimulated by PHT (20, 100 mg/L), bone morphogenetic protein-2 (BMP-2) concentration was analyzed by enzyme linked immunosorbent sandwich assay (ELISA). RESULTS: At the concentration of 20 or 100 mg/L, PHT significantly enhanced the proliferating activity and ALP activity of hPDLF (P<0.01). PHT at 100 mg/L could increase protein synthesis of hPDLF (P<0.05). The capability of mineralization and BMP-2 expression of hPDLF were increased significantly (P<0.01) in 100 mg/L group when compared with that in the control group. However, higher concentration (2 500 mg/L) not only changed cell morphology, but also significantly inhibited cell activity. CONCLUSION: The results suggested that proper doses of PHT could promote proliferation and biosynthesis and also enhance osteogenesis by increasing the differentiation, mineralization and BMP-2 expression of hPDLF while higher concentrations of PHT had cytotoxic effect.
OBJECTIVE: To study the biological effects of phenytoin (PHT) on cultured human periodontal ligament fibroblasts (hPDLF), and explore the possibility of its accelerating periodontal regeneration. METHODS: Increasing concentrations of PHT (1, 5, 20, 100, 500, 2 500 mg/L) were added into the medium of the fourth passage of cultured hPDLF, respectively. After co-incubated for 3 days, cell proliferation activity, the total amount of protein and alkaline phosphatase (ALP) activity were detected. Mineralized sodium and PHT (20, 100, 500 mg/L) were added into the medium of the fourth passage hPDLF. After co-incubated, the mineralized nodules formation were detected by Von Kossa staining. The third passage hPDLF were stimulated by PHT (20, 100 mg/L), bone morphogenetic protein-2 (BMP-2) concentration was analyzed by enzyme linked immunosorbent sandwich assay (ELISA). RESULTS: At the concentration of 20 or 100 mg/L, PHT significantly enhanced the proliferating activity and ALP activity of hPDLF (P<0.01). PHT at 100 mg/L could increase protein synthesis of hPDLF (P<0.05). The capability of mineralization and BMP-2 expression of hPDLF were increased significantly (P<0.01) in 100 mg/L group when compared with that in the control group. However, higher concentration (2 500 mg/L) not only changed cell morphology, but also significantly inhibited cell activity. CONCLUSION: The results suggested that proper doses of PHT could promote proliferation and biosynthesis and also enhance osteogenesis by increasing the differentiation, mineralization and BMP-2 expression of hPDLF while higher concentrations of PHT had cytotoxic effect.