Determination of Anti-aging Effect of Comfrey Cell Suspension Culture Extract on Human Normal Fibroblast Cells

Abstract

This study was a purpose to investigate the anti-aging activity of Comfrey cell suspension culture (CCSC) extract. The effects of CCSC extract on human normal fibroblast cells (CRL-2076) were examined. Comfrey callus and suspension culture were obtained in Murashige and Skoog (MS) medium including 1 mg/L 2,4-Dichlorophenoxyacetic acid (2.4-D). Total phenolic contents were determined in CCSC lyophilizes, the highest phenolic content was found to be 1181 µM gallic acid equivalent in the extract prepared with 70% ethanol solvent. The half-life concentration (IC50 value) of the extract was determined as 83 µg/ml with 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. It was found that CCSC extract incubated for 48 h did not reduce cell viability on CRL-2076 cells at doses of 100 and 200 µg/mL. The addition of the CCSC extract at 100 µg/mL concentration to aged CRL-2076 cells increased the collagen type 1 alpha 1 (COL1A1) gene expression level 1.2-fold, elastin (ELN) gene expression level 1.4-fold, and hyaluronan synthase 3 (HAS3) gene expression level 1.4-fold. However, the addition of the CCSC extract at 200 µg/mL concentration to aged CRL-2076 cells increased the COL1A1 gene expression level 1.3-fold, ELN gene expression level 2.7-fold, and HAS3 gene expression level 1.7-fold. In the study, CCSC extract was shown to have anti-aging properties on CRL-2076 cells, suggesting that CCSC extract can be used as active raw material in natural anti-aging cosmetic products.