86 ANTI-POLLUTION * 150 **
100
50
0 Unstressed 0.5% HerbaShield URB
Unstressed +
BaP 0.5% HerbaShield URB
BaP +
Figure 6: The active ingredient empowers the cellular detoxification machinery. Human keratinocytes were stressed for 48h with 0.05 μg/ml BaP or not (unstressed). Nrf2 activity was significantly enhanced upon treatment with the multi-component active ingredient in a concentration of 0.5%. N = 4; Mean ± SD; Student’s unpaired t-test versus untreated; * = p <0.05; ** = p <0.01.
Results The treatment with the active ingredient stimulated Nrf2 activity, indicating that the extract can induce the skin’s endogenous detoxification machinery (Fig 6). The active ingredient provided additional detoxification power. Cells stressed with BaP showed higher Nrf2 activity than unstressed cells, implying that the cellular detoxifying machinery is reinforced (Fig 6).
Reducing antioxidative stress - enhancing Phase I detoxification Objective To show that the active ingredient provides additional detoxification power and counteracts pollutant-induced skin stress.
Technique - DCF Assay A non-fluorescent precursor of a fluorescent dye diffuses into keratinocyte cells, where
a 3 *** *** 2 *** 2 *** 1 *** 1
unstressed
5 µg/ml BaP
5 µg/ml BaP + 0.6% HerbaShieldURB
Figure 8: The active ingredient protects against pollutant-induced oxidative stress. Unstressed cells produce low amounts of free radicals as shown by a low fluorescence signal (left panel - unstressed). Cells incubated with BaP generate oxidative stress as shown by a high fluorescence signal (center panel - 5μg/ml BaP). The additional presence of the active ingredient completely relieves oxidative stress (right panel - 5μg/ml BaP + active ingredient), as shown by the absence of fluorescence. Upper images show fluorescence signals, lower images show phase contrast images. Scale bars = 50 μm.
radical oxygen species convert the precursor into the fluorescent dye. The enhanced fluorescence signal is monitored and taken as a measure of oxidative stress. Pre- incubation with the active ingredient for 24 h followed by 1 h incubation with 1 mM H2
O2
2.5 μg/ml BaP, or 100 μg/ml particulate matter (PM) in the presence of the active ingredient.
Results
Incubation with particulate matter (PM) or BaP led to massive oxidative stress. The formation of intra-cellular reactive oxygen species (ROS) increased threefold and was even higher as compared to incubation with 1 mM H2
O2 . The active ingredient was able
to entirely inhibit the pollutant-induced formation of ROS (Fig 7). The reduced quantity of radicals was confirmed by fluorescence microscopy (Fig 8).
b 3 * *** *** ,
Pollution causes skin ageing - an in vivostudy Objective To study the anti-pollution effect of the active ingredient and to evaluate anti-age parameters, such as skin firmness, skin elasticity, skin roughness and wrinkles in a double-blind, placebo-controlled, randomised in vivo study, with 2x21 Caucasian female volunteers, between 30– 65 years of age, living in an urban area, and smoking at least 5 cigarettes per day. One group applied a cream formulation containing 1% active ingredient to their face, twice a day, for 4 weeks. The other group applied the same formulation without the active ingredient (placebo).
0 – – Unstressed H2 – 0.06 0.15 0.3 0.6 HerbaShield URB (%) 02 PM PM+HerbaShield URB Unstressed H2 0 – – – 0.06 0.15 0.3 0.6 HerbaShield URB (%) 02 PM PM+HerbaShield URB
Figure 7: The active ingredient counteracts the effects of particulate matter and pollutant-induced skin stress. Human keratinocytes (HaCaT cells) were stressed for 1 h with 100 μg/ml particulate matter (A; PM), and 2.5 μg/ml BaP (B), respectively. Treatment with the extract significantly reduced the stress response. N = 4; Mean ± SD; Student’s unpaired t-test versus stressed (PM or BaP) but untreated; * = p < 0.05; *** = p < 0.001.
PERSONAL CARE EUROPE
Techniques (1) Skin firmness and elasticity was measured by cutometry determining R0 and R2, respectively. (2) Skin roughness and wrinkles were determined evaluating the RZ (a marker for skin roughness and fine lines) and wrinkle volume by Primos 3D. In addition, an expert dermatologist scored these parameters by using the 8-degree Japanese Cosmetic Industry Association’s wrinkle grading system. (3) Skin complexion was measured using VISIA-CR, a standardised imaging system, taking high-resolution photos and allowing the calculation of brownish and reddish skin irregularities. Moreover, an expert dermatologist evaluated skin tone, spots, evenness, and complexion on a 5-degree scale. (4) Skin oiliness, the so-called lipidic index, was evaluated using sebumeter tape and grease spot photometry. (5) Changes in epidermal barrier function were evaluated by measuring transepidermal water loss (TEWL). Finally, the in vivo test included a self- assessment using a questionnaire completed before and after the study. Results are shown in Figures 9-12.
April 2018
Oxidative Stress (normalized DCF Fluorescence) Nrf2 Activity (%)
Oxidative Stress (normalized DCF Fluorescence)
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