ABSTRACT
The purpose of present study was to investigate the effects of physiologically active compound (SD62-122) from Phlomidis Radix on the cell cycle progression and its molecular mechanism in human gingival fibroblasts(HGFs). For this purpose, fibroblasts were isolated and cultured from excisioned gingiva during crown lengthening procedure in healthy adult. The following parameter were evaluated that there are cell number counting, MTT assay, cell cycle progression, western blot analysis. The cell number and MTT assay of primary cultured fibroblast was not increased at 2 days but significant increased compare to negative control at 3days(p<0.05). S phase was increased and G1 phase decreased in both 10(-8)M and 10(-9)M of SD62-122 in cell cycle analysis. The cell cycle regulation protein levels of Cyclin D1, Cyclin E, cdk 2, cdk 4 and cdk 6 were increased compare to control in both 10(-8)M and 10(-9)M of SD62-122. The protein levels of p21 and p53 were decreased compare to control, but the level of pRb was not changed compare to control in 10(-9)M of SD2-122. These results suggested that physiologically active compound (SD62-122) isolated from Phlomidis Radix increases the cell proliferation and cell cycle progression in HGFs, which is linked to increased cell cycle regulation protein levels of Cyclin D1, Cyclin E, cdk 2, cdk 4 and cdk 6, and decreased the levels of p21, p53.
Subject(s)
Adult , Humans , Blotting, Western , Cell Count , Cell Cycle , Cell Proliferation , Crown Lengthening , Cyclin D1 , Cyclin E , Cyclins , Fibroblasts , G1 Phase , Gingiva , S PhaseABSTRACT
Osteoblasts from alveolar bone may have an important role in the bone regeneration for periodontium, but their culture and characterization are not determined yet. The purpose of this study was to investigate the biological characteristics of primary explant cultured osteoblasts(PECO) from alveolar bone. Osteoblasts were isolated and cultured from alveolar socket of extracted tooth in children. To compare the characteristics, osteoblasts and gingival fibroblasts were cultured with DMEM at 37degrees C, 5% CO2, 100% humidity incubator, and human fetal osteoblasts cell line(hFOB1) were cultured with DMEM at 34degrees C, 5%, CO2, 100% humidity incubator. To characterize the isolated bone cells, morphologic change, cell proliferation and differentiation were measured. Morphology of PECO was small round body or cuboidal shape on inverted microscope and was similar with hFOB1. PECO became polygonal shape with stellate and had an amorphous shape at 9th passage in culture. PECO had significantly higher activity than that of gingival fibroblasts and hFOB1 in alkaline phosphatase activity. The expression of osteocalcin and bone sialoprotein in PECO was notably increased when compared with hFOB1 and gingival fibroblasts. These result indicated that PECO from alveolar bone in children has an obvious characteristics of osteoblast, may be applied for the regeneration of bone.
Subject(s)
Child , Humans , Alkaline Phosphatase , Bone Regeneration , Cell Proliferation , Fibroblasts , Humidity , Incubators , Integrin-Binding Sialoprotein , Osteoblasts , Osteocalcin , Periodontium , Population Characteristics , Regeneration , ToothABSTRACT
Nicotine is one of the major components of cigarette smoking which causes various systemic and local diseases to human body. The purpose of the present study was to investigate the effects of nicotine on bone mineralization in human fetal osteoblasts cell line(hFOB1). To compare the alkaline ph-osphatase(ALP) synthesis, hFOB1 were cultured with DMEM/F-12 1:1 Mixture and 100 pg/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1 microgram/ml, 10 microgram/ml, 100 microgram/ml of nicotine. And to compare the calcium accumulation, hFOB1 cultured for 23 days were quantified and photographed. ALP activity of hFOB1 exposed to nicotine was not significantly changed at a lower concentrations of nicotine, but was significantly decreased at a higher concentrations (10 microgram/ml, 100 microgram/ml) of nicotine (p<0.05). A quantified calcium acculation in hFOB1 was significantly decreased at 1, 10, and 100microgram/ml of nicotine (p<0.05). Significantly decreased calcium deposition was observed at 1, 10, and 100microgram/ml of nicotine. These results indicate that a higher concentration of nicotine show a negative effects on mineralization of hFOB1.