The Effect of Anti-aging Peptides on Mechanical and Biological Properties of HaCaT Keratinocytes

Tomasz Kobiela,Ida Dulinska-Molak,Wojciech Święszkowski,Irena Eris,Małgorzata Milner-Krawczyk,Konstancja Bobecka-Wesołowska,Monika Pasikowska-Piwko

doi:10.1007/s10989-017-9648-7

Abstract

Atomic force microscopy (AFM) and fluorescence microscopy was applied to determine the influence of the anti-aging peptides on the morphology and the mechanical properties of keratinocytes. Immortalized human keratinocytes (HaCaT) were treated with two anti-aging bioactive peptides: Acetyl Tetrapeptide-2 and Acetyl Hexapeptide-50 (Lipotec). The AFM measurement of the keratinocyte stiffness were carried after 48 h exposure at an indentation depth of 200 nm. AFM analysis showed increase of the cell stiffness for cells treated with Acetyl Tetrapeptide-2 (P1) in concentration range. Acetyl Hexapeptide-50 (P2) at concentration of 0.05 µg/ml also increased the stiffness of HaCaT cells but at higher concentrations 0.5 and 5 µg/ml cell stiffness was lower as compared to untreated control. Fluorescence microscopy revealed remodeling of actin filaments dependent on the concentration of P2 peptide. The mechanical response of HaCaT cells treated with P2 peptide corresponds to change of transcription level of ACTN1 and SOD2 which activity was expected to be modulated by P2 treatment.

Highlights

In response to still growing demands, the researchers working in pharmaceutical and cosmeceutical fields are developing new, more efficient, bioactive and biocompatible antiaging compounds
First group - signal peptides generally stimulate protein production including the components of extracellular matrix (ECM) like collagen, elastin, glycosaminoglycans and fibronectin
We investigated the influence of two anti-wrinkle bioactive peptides on mechanical properties of HaCaT keratinocytes

Summary

Introduction

In response to still growing demands, the researchers working in pharmaceutical and cosmeceutical fields are developing new, more efficient, bioactive and biocompatible antiaging compounds. First group - signal peptides generally stimulate protein production including the components of extracellular matrix (ECM) like collagen, elastin, glycosaminoglycans and fibronectin. Signal peptides stimulating activity of fibroblasts resulting in ECM collagen production result in firmer and younger look of the skin. Mechanical properties of the skin change with aging and under influence of various external factors like UV irradiation and environmental chemical pollution (Sobiepanek et al 2016). This leads to a disturbance in synthesis of collagen and other structural ECM and cellular proteins causing the loss of skin firmness and elasticity. We investigated the influence of two anti-wrinkle bioactive peptides on mechanical properties of HaCaT keratinocytes. The sample absorbance was determined at 595 nm wavelengths in a microplate reader (Synergy H4, Biotek) The results are expressed as a percent of untreated control and are an average of three experiments

Materials and Methods

Results and Discussion

Conclusions