Curcumin Promotes Collagen Type I, Keratinocyte Growth Factor-1, and Epidermal Growth Factor Receptor Expressions in the In Vitro Wound Healing Model of Human Gingival Fibroblasts

Auspreeya Rujirachotiwat, Supaporn Suttamanatwong, Auspreeya Rujirachotiwat, Supaporn Suttamanatwong

Abstract

Objective: Curcumin promotes oral wound healing; however, the underlying mechanism remains unknown. We hypothesized that curcumin may regulate gene expression in human gingival fibroblasts (hGFs). This study investigated the effect of curcumin on the expression of wound healing-related genes, collagen type I (COL1), keratinocyte growth factor (KGF)-1, and epidermal growth factor receptor (EGFR), in the in vitro wound healing model of hGFs, as well as the signaling pathway involved in the regulation of these genes by curcumin.

Materials and methods: The hGFs were treated with curcumin in the unwounded condition and in the in vitro wound healing model (scratch assay). Gene expression was determined by quantitative polymerase chain reaction. PD98059 was used to elucidate whether extracellular signal regulated kinase (ERK) signaling is involved in the curcumin-regulated gene expression in hGFs. Cell migration was also analyzed by the scratch assay.

Statistical analysis: Data were analyzed by independent t-test or one-way analysis of variance (ANOVA) followed by Tukey's Honestly Significant Difference ( HSD) test.

Results: In unwounded hGFs, curcumin significantly increased KGF-1 and EGFR expressions but not COL1 mRNA expression. Interestingly, curcumin significantly upregulated COL1, KGF-1, and EGFR expressions in the in vitro wound healing model. Furthermore, PD98059 significantly decreased the curcumin-induced COL1 and EGFR expressions, but did not significantly affect KGF-1 upregulation by curcumin. However, hGF migration was not affected by curcumin treatment.

Conclusion: Curcumin induced KGF-1 and EGFR expressions in unwounded hGFs. In the in vitro wound healing model, curcumin upregulated COL1 and EGFR expression via the ERK pathway and increased KGF-1 expression, possibly by an ERK-independent mechanism.

Conflict of interest statement

The study protocol was approved by the Ethics Committee of the Faculty of Dentistry, Chulalongkorn University.None declared.

European Journal of Dentistry. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Figures

A
A
Fig. 1 The mRNA expression of COL1 ( ), KGF-1 (B), and EGFR (C) in hGFs in response to curcumin. Cells were treated with varying concentrations of curcumin or DMSO for 24 hours. The level of gene expression was determined by qPCR. The data are the mean ± SD from three independent experiments in which each group has samples from three different patients (n= 9). Statistical analysis was performed by one-way ANOVA followed by Tukey’s HSD test. *indicates compared with the control group (p< 0.05). ANOVA, analysis of variance; COL1, collagen type I; EGFR, epidermal growth factor receptor; hGFs, human gingival fibroblasts; HSD, honestly significant difference; KGF 1, keratinocyte growth factor-1; qPCR, quantitative polymerase chain reaction; SD, standard deviation.
Fig. 2
Fig. 2
The expression of COL1 (A), KGF-1 (B), and EGFR (C) in hGFs in response to curcumin in thein vitrowound healing model. The hGF monolayer was scratched using a sterilized pipette tip and then treated with 1 µM of curcumin or DMSO for 24 hours. The level of gene expression was determined by qPCR. The data are the mean ± SD from three independent experiments in which each group has samples from three different patients (n= 9). Statistical analysis was performed by independentt-test. *indicates compared with the control group (p< 0.05). COL1, collagen type I; EGFR, epidermal growth factor receptor; hGFs, human gingival fibroblasts; KGF 1, keratinocyte growth factor-1; qPCR, quantitative polymerase chain reaction; SD, standard deviation.
Fig. 3
Fig. 3
Effect of inhibiting ERK signaling on the curcumin-induced COL1 (A), KGF-1 (B), and EGFR (C) expressions in hGFs in thein vitrowound healing model. The hGF monolayer was scratched and treated with 100 nM PD98059 for 1 hour prior to 1 µM curcumin for 24 hours. The level of gene expression was determined by qPCR. The data are the mean ± SD from three independent experiments in which each group has samples from three different patients (n= 9). Statistical analysis was performed by one-way ANOVA followed by Tukey’s HSD test. *indicates compared with the control group (p< 0.05), #indicates compared with curcumin-treated group (p< 0.05). ANOVA, analysis of variance; COL1, collagen type I; EGFR, epidermal growth factor receptor; hGFs, human gingival fibroblasts; HSD, honestly significant difference; KGF 1, keratinocyte growth factor-1; qPCR, quantitative polymerase chain reaction; SD, standard deviation.
Fig. 4
Fig. 4
Effect of curcumin on gingival fibroblast migration. The hGF monolayer was scratched and then treated with DMSO (A–C) or 1 µM curcumin (D–F) for 24 to 48 hours. The images are representative of three separate experiments. (G) The remaining wound area at 0, 24, and 48 hours was analyzed by Image-Analysis J Software. The data are the mean ± SD from three independent experiments in which each group has samples from three different patients (n= 9). Statistical analysis was performed by independentt-test. hGFs, human gingival fibroblasts; SD, standard deviation.

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