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.

Key words: phenytoin, periodontal ligament fibroblasts, cell culture, mineralization