Effect of Collagen-Related compound on cell proliferation and matrix production with cultured human dermal fibroblasts

Abstract

Collagen, an important component of the extracellular matrix and produced by dermal fibroblasts, is the key to healthy and firm skin. However, the ability of the fibroblast to produce a sufficient amount of collagen is depleted with age. Collagen hydrolysate (CH) is the product of extensive degradation of gelatine. In the present study, CH of molecular ranging 1-5kDa and its related compounds were evaluated for their effect on cell proliferation and matrix production of cultured human dermal fibroblasts (HDF). Two age classes of HDF were prepared, namely the normal HDF and senescence acceleration HDF, induced by hydrogen peroxide (50μM) treatment. CH was added to both classes at different concentrations (1μg/mL, 10μg/ml, 100μg/ml and 1000μg/ml). The cell proliferation was measured using WST-1 assay and the matrix production was evaluated by alcian blue (AB) pH 2.5. An additional assay, the β-galactosidase activity assay (SA-β-Gal), was measured to evaluate the degree of cellular senescence. The cell proliferation was measured using WST-1 assay and matrix production was measured with alcian blue (AB) pH2.5. The results revealed that cell proliferation had significant differences in both normal HDF and senescence-induced HDF. Beta-Galactosidase results revealed a positive decline of senescent cells at higher concentrations of CH (100 & 1000 μg/L). The Alcian Blue pH2.5 showed contrasting results in normal HDF (p<0.05) at 100 & 1000 μg/L and senescence-induced HDF showed no significant differences. This led to the conclusion that a higher CH concentration is able to produce significant differences in both normal and senescence HDF on cell proliferation and matrix production.