74 LIFESTYLE COSMETICS
Waking up the skin in the morning Dawnergy™ peptide (INCI: Water (Aqua), Sodium Phosphate, Nonapeptide-1, Disodium Phosphate, Caprylyl Glycol) is a nonapeptide that can wake up the skin cells through the modulation of JARID1a and clock genes and can also boost energy production by mitochondria. Different in vitro and in vivo tests were performed to evaluate the efficacy of the active ingredient.
Effect on JARID1a and clock genes l Setting the cellular alarm clock earlier in the morning
Human keratinocytes were synchronised to day-night rhythms by temperature changes and treated with Nonapeptide-1 for 48 hours. Then, at time points, corresponding to the early morning (7:30 a.m.) and the late morning (11:30 a.m.), different measurements were made. Proteins were extracted from the cell’s nucleus for the determination of JARID1a levels by an ELISA assay and mRNA was isolated and used to obtain gene expression profiles of clock genes by Real Time PCR Arrays. At 7:30 a.m., with the peptide treatment
protein levels of JARID1a resembled the levels obtained in control at 11:30 (Fig 1). Consistent with the fact that JARID1a drives the activity of the internal cellular clock in the morning, an increase in expression of the clock gene Per3 at 7:30 a.m. was also observed, reaching levels observed later (at 11:30 a.m.) in the control. l Reversing the decline occurring with ageing Human keratinocytes from a 26-year-old (y.o.) donor and from a 54-y.o. donor were synchronised to day-night cycles by temperature changes. The 54 y.o. cells were incubated with Nonapeptide-1 for 48 hours. Finally, gene expression of Per3 was analysed by means of Real Time PCR Arrays. The analysis of cells from donors of
different ages revealed significant decrease of Per3 expression with age. Treatment of older keratinocytes with the peptide significantly increased levels of this ageing- sensitive clock gene (Fig 2).
250 200 150 100 ** 50 0 26
y.o.Control 54 y.o. Control 54 y.o. + DawnergyTM peptide
Figure 2: Gene expression of Per3 in younger and older skin cells (**p<0.01 vs 26 y.o. control; *p<0.05 vs 54 y.o. control).
Control Nonapeptide-1 *
Figure 3: Fluorescence microscopy images of cells (red shows high mitochondrial energy state).
Effect on mitochondria l Mitochondrial energy increase Human dermal fibroblasts were treated with Nonapeptide-1 for 16 hours. The mitochondrial membrane potential, which is related to the production of ATP, was determined by means of a dye-mediated detection assay. Cells were imaged using confocal fluorescence microscopy for the quantification of energised cells. Treatment of skin cells with the active
ingredient increased the mitochondrial membrane potential, which is linked to an enhanced mitochondrial function and ATP
Complex I activity
120 100 80 60 40 20 0
Control ** **
160 140 120 100 80 60 40 20 0
Metformin DawnergyTM peptide Figure 4: Relative values of mitochondrial complexes activity (**p<0.01 vs control). PERSONAL CARE ASIA PACIFIC September 2019 Control
synthesis (Fig 3). In addition, this energising effect was not associated with an increase in the levels of reactive oxygen species (ROS) (data not shown). l Neutralising the passage of years in mitochondrial function Human dermal fibroblasts were incubated with Nonapeptide-1. As a negative control the cells were treated with 10 mM metformin. As a complex II inhibition control, the cells were treated with 30 mM 2-Thenoyltrifluoroacetone (TTFA). After 48 and 24 hours of treatment respectively, activities of mitochondrial complexes I and
Complex II activity **
**
TTFA
DawnergyTM peptide
Relative complex I activity (%)
Per3 relative gene expression (%)
Relative complex II activity (%)
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