In vitro Effect of L-ascorbic acid on gingival fibroblast to regulate wound healing.

Abstract

Background and Objective: Besides closing the wound by intrinsic properties like migration and proliferation, gingival fibroblasts produce a major source of extracellular matrix especially collagen and play a major role in oral wound healing. Vitamin C or L-ascorbic acid has diverse functions in the body, including an essential role in hydroxylation reactions which is necessary for collagen formation. Whether L-ascorbic acid can promote gingival wound healing through inducing proliferation of fibroblasts is of our interest. The aim of this study is to evaluate the effect of L-ascorbic acid on gingival fibroblasts behaviors to promote wound healing in vitro.

Materials and Methods: Primary human gingival fibroblasts isolated from gingival tissue from healthy volunteers were rinsed 3 times a day with medium containing L-ascorbic acid of various concentrations. Local effect of L-ascorbic acid rinsing on in vitro wound healing was assessed by mean of Scratch-test assay. Cell migration, cell proliferation and extracellular matrix production were analyzed by transwell-migration assay, MTT assay and real-time RT-PCR respectively.

Result: Rinsing with vitamin C at concentration of 10 and 20 µg/ml accelerated fibroblast migration in transwell-migration assay but no significant effect on cell proliferation and in vitro wound closure. However, rinsing with vitamin C at higher concentration (50 µg/ml) significantly delayed wound closure comparing with the control group (p value=0.05). This data was in accordance with cell viability assessed by MTT assay demonstrating that Vitamin C at concentration above 50 µg/ml significantly reduced fibroblasts proliferation. However, real time PCR demonstrated that vitamin C at 50 µg/ml significantly increased expression of COL1, FN, IL-6 and bFGF, molecules related to wound healing.

Conclusion: Vitamin C at various concentrations differentially affect wound healing capability of gingiva fibroblast. Vitamin C is safe and can be prescribed to patients after oral surgery. However, suitable duration and appropriate drug administration method should be adjusting to maximize it benefit and further clinical study is required.