Protective effects of tetrahydrocurcumin (THC) on fibroblast and melanoma cell lines in vitro: it's implication for wound healing

Abstract

The aim of this study was to investigate the role of tetrahydrocurcumin (THC) against various skin health parameters using in vitro human foreskin fibroblast and melanoma cell lines (i.e. HFF-1 and B16-F10). The study was assessed using cell viability by MTT assay, identification of extracellular matrix component in HFF-1 cell line (i.e. collagen, elastin and hyaluronic acid), melanin synthesis in B16-F10 cells, cell viability against UVB-induced stress in HFF-1 cells, and in vitro wound healing by the scratch assay. THC was found to be safe and nontoxic up to the concentration of 10 µg/mL with improved level of collagen (37.90%), elastin (90.1%), and hyaluronic acid (74.19%) at 1 µg/mL. Besides, melanin was significantly inhibition by 78.5% at the lowest THC concentration of 0.1 µg/mL. UVB-protection rate was significantly improved by 61.2% and improved cell viability by THC in HFF-1 cells, which indicated protection from photoaging. In addition, THC showed significant wound healing activity (78.51%) and greater migration of fibroblast in HFF-1 cells at different time period. It can be concluded from the study that THC can protect the skin matrix with improved extracellular component synthesis and would healing via collagen synthesis in the skin, which improved the skin elasticity and tightness. Overall, it might be suggested that THC can be used as a safe skin whitening agent, wounds management, cosmetic applications, and treating various skin-related disorders.

Keywords: B16-F10; Extracellular matrix; HFF-1; Hyaluronic acid; Scratch assay; Tetrahydrocurcumin.

Fig. 1

Concentration-dependent effects of THC on human foreskin fibroblast (HFF-1) cell line for extracellular matrix component, collagen. **p ≤ 0.01 versus vehicle control and ^## p ≤ 0.01 versus ascorbic acid (using Student’s t-test with three independent assay)

Fig. 2

Concentration-dependent effects of THC on human dermal fibroblast cells (HFF-1) cell lines for extracellular matrix component, elastin. ***p ≤ 0.001 and **p ≤ 0.01 versus vehicle control and ^## p ≤ 0.01 versus positive control (ascorbic acid) group using one way ANOVA (using Tukey’s test with three independent assay)

Fig. 3

Synthesis of extracellular matrix component, hyaluronic acid by THC against UVB-induced stress in human foreskin fibroblasts (HFF-1) cell lines. ***p ≤ 0.001 and **p ≤ 0.01 all statistical comparison with respect to vehicle control using one-way ANOVA (using Tukey’s test with three independent assay)

Fig. 4

Inhibitory effect of THC on melanogenesis (skin whitening potential) in mouse melanoma (B16-F10) cell line. ***p ≤ 0.001 versus vehicle control and ^### p ≤ 0.001 versus positive control (ascorbic acid) using one-way ANOVA (using Tukey’s test with three independent assay)

Fig. 5

Anti-wrinkling potential and cytoprotective effect of THC against UVB-induced stress in human foreskin fibroblasts (HFF-1) cell lines. L-AC L-ascorbic acid, THC tetrahydrocurcumin. ***p ≤ 0.001, **p ≤ 0.01 and *p ≤ 0.05; a and b represents statistical comparison with vehicle control and ascorbic acid group respectively using one way ANOVA (using Tukey’s test with three independent assay)

Fig. 6

Representative pictures show the migration of HFF-1 cells after induction of a scratch. All the pictures were taken immediately after the scratch was induced (i.e. at 0 h), after 12 h (12 h), and 48 h (48 h). The fibroblast on the pictures were cultured in conditioned media. Pictures are taken at 50 times magnification. THC tetrahydrocurcumin, DMSO dimethyl sulfoxide (vehicle control). ***p ≤ 0.001 and **p ≤ 0.01 represents statistical comparison with vehicle control using Student’s t-test with three independent assay