Vitamin C or L-ascorbic acid has diverse functions in the body, especially healing promotion in tissue injury via participating in the hydroxylation reactions required for collagen formation. Systemic administration of vitamin C plays an important role on gingival fibroblast proliferation and functions. Whether local or rinsing administration of vitamin C alters gingival fibroblast wound healing behavior remains unclear. Therefore, it is of interest to investigate the effect of vitamin C on gingival fibroblast behavior utilizing cell culture. Primary human gingival fibroblasts isolated from gingival tissue were rinsed with medium containing various concentrations of vitamin C. Cell migration, cell viability was assessed using MTT assay. The viability assay showed >95% live cells, and no significant (P > 0.05) difference in these values was observed at different concentrations at 24 hrs. The levels of cell proliferation were not significantly different among the control and experimental groups in 24 hrs experimental time-points (p > 0.05). Vitamin C is nontoxic and can be used for further experiments to validate for clinical application. This was further confirmed with morphological examination after 24hrs incubation on control and experimental group drugs observed under the phase contrast microscope.