In vitro protective effects of Paeonia mascula subsp. hellenica callus extract on human keratinocytes

Abstract

Natural ingredients have been used to improve the state of health in humans. The genus Paeonia has been studied only limited yet it's reported to have many activities such as antioxidant and anti-inflammatory. To this context, here we focused on an endemic Paeonia species in Attica. This study aims to present the development of the Paeonia mascula subsp. hellenica callus extract and its pleiotropic bioactivity on human primary keratinocytes exploring its potential application as an active agent in skin-related products. This extract showed a high scavenging activity with high phenolic content and an interesting metabolic profile. At a molecular level, the study on the transcript accumulation of genes revealed that this extract exhibits in vitro skin-related protection properties by mediating mitochondrial energy, cell proliferation, immune and inflammatory response and positively regulates genes involved in epidermal and in stratum corneum function. Besides, the extract is proven not skin irritant on reconstructed human skin model. These findings indicate that the specific P. mascula subsp. hellenica extract possesses significant in vitro protection activity on human epidermis and provides new insights into its beneficial role in skin confirming that the advent of biotechnology contribution the past few decades.

Figure 1

Initiation and growth of callus cell lines. Embryo formation (A, B), callus formation (C, D), callus with first stage globular embryos (E) growth of callus as small scale suspension cultures (F).

Figure 2

Represented chromatograms at 280 and 360 nm for elicitated (green) and unelicitated (red) cells.

Figure 3

OCR levels expressed as mean ± SEM for: control (untreated NHEK cells), NHEK cells treated with POCE (POCE), NHEK cells treated with H₂O₂ (H₂O₂) NHEK cells pre-treated with POCE and after with H₂O₂ (POCE + H₂O₂). *p < 0.05 significantly different from the control (ANOVA test).

Figure 4

Cell viability levels expressed as mean ± SEM based on reconstructed human skin model for: (A) reconstructed human skin model treated with POCE, phosphate-buffered saline (DPBS) (positive control, PC) and 5% Sodium Dodecyl Sulfate (SDS) (negative control, NC). (B) reconstructed human skin model treated with a cosmetic formulation with POCE and 1% triton-100 as negative control (NC). *,**p < 0.05 significantly different from the formulation point.

Figure 5

Relative gene expression of: CDSN, TJP1, CAV1, PXN, OCLN, CMAR, ITGA1, KLK7, INHBA, SLC27A3, UGCG, ABCA12, GBA1, DEFB4, IL1α, IL1β, IL6,IL8,TNF-α expressed as a fold change ± SEM compared to the control NHEK cells and using ACTB and GADPH as internal reference genes. The experimental conditions were control cells, cells treated with POCE (0.05 μg/ml) (POCE). *p < 0.05 indicates groups significantly different from the control (ANOVA test). The data correspond to the mean ± SEM of three independent experiments and six replicates each time.