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Objective: The purpose of this study was to investigate the effect of schizophyllan on cytotoxicity proliferation and migration of human gingival fibroblast (HGF).
Materials and Methods: For cytotoxicity assay : HGF were plated in 96 well-plate with 2×104 cells/ml and cultured in DMEM solution containing 10% FBS and antibiotics/antimycotics; treated with schizophyllan at final concentration 0.1, 0.5, 1.0, 5.0 and 10.0 mg/ml and incubated for 24 hours in CO2 incubator at 5% CO2, 100% humidity. Cell viability was evaluated using MTT assay.
For cell proliferation assay : HGF were inoculated in 96-well plate with 1×105 cells/ml for 24 h. Afterward schizophyllan at final concentration of 0.1, 0.5, 1.0, 5.0 and 10.0 mg/ml were added and cultured for 13 days under same condition as described above. The media were routinely changed every 2 days. Numbers of viable cells were determined using MTT assay.
For cell migration assay : HGF at concentration of 1×105 cells/well were seeded into for 24 well plate and grown for 24 hours in DMEM containing 10% FBS and antibiotics/antimycotics. Cell were scratched by using 1,000 ul steriled pipette tip. The schizophyllan (0.1, 0.5, 1.0, 5.0 and 10.0 mg/ml) was added and further incubated under same condition for 24 hours. After 24 hours cells were stained with toluidine blue O and left dried overnight. The migrated cells were counted by using Image Pro Plus program (version 7).
Result: The result suggested that shizophyllan had no toxicity in all concentrations tested. The cell proliferation assay showed that when cells were treated with 10.0 mg/ml schizophyllan, the amount of cell proliferation was 5.7×104 cell/well which is higher than control (5.0×104 cell/well). There were significant differences when comparing the amount of cell proliferation between schizophyllan at the concentration of 10 mg/ml with the control. The cell migration assay showed that schizophyllan at 5.0 and 10.0 mg/ml can promote the migration of cells into wounded area. The mean of migrated cell was 167, 204 cells/Sq mm respectively. There were significant differences in the mean of migrated cells between the concentration at 5.0 and 10.0 mg/ml with the control
Conclusion: Schizophyllan up to 10 mg/ml has no cytotoxicity to HGF and can promote cell proliferation. Schizophyllan at 5 and 10 mg/ml can induce the migration of cell into the wounded area. This study may be useful for the development of some drugs to promote wound healing in the oral cavity.
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