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Ectoin® for visible light protection

EX VIVO study
(human living skin explants)

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Visible light – definition
• Visible light is the portion of the electromagnetic spectrum that is visible to the human
eye.
• Wavelength 390-700nm, visible light includes blue light (420-440nm).
• In the last years, "visible light" (39% of the sun radiation) was discovered to be damaging
for the skin tissue. Visible light induces DNA damage as it produces DNA single strand
breaks and intracellular reactive oxygen species (ROS).

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EX VIVO Study design

• Human living skin explants (18-year-old Caucasian woman)


• Topical treatment (2 μl/explants) with placebo and Ectoin® on Day 0, D2, D3
and D4.
• Exposure to visible light (65 J/cm2) on D4.
• Sampling of skin explants on D5.
• Evaluated parameters: immunostaining of Nrf2 and MC1R.

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Background information: Nrf2

• Nrf2 = key transcription factor in the cellular response to oxidative stress.


• Is important for protecting human skin keratinocytes from UVA and
pollutant-induced damage.
• Under oxidative stresses, including UV irradiation, Nrf2 is activated by
phosphorylation and translocates from the cytoplasm to the nucleus (Fig. 1).
• In the nucleus, Nrf-2 binds to the DNA at the location of the Antioxidant
Response Element (ARE) or also called hARE (Human Antioxidant Response
Element)  master regulator of the total antioxidant system

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Fig1: Schematic representation of Nrf2- activated response under oxidative stresses.
Background information: MC1R

• MC1R = melanocortin 1 receptor; marker for pigmentation


• also known as melanocyte-stimulating hormone receptor (MSHR) and melanin-
activating peptide receptor
• It is located on melanocytes, which produce the pigment melanin through the
process of melanogenesis.
• Inactivation of MC1R leads to a decrease of melanin production.
• MC1R signaling is also a potent anti-inflammatory pathway and has been
shown to promote antimelanoma immunity.

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Immunostaining

• Immunostaining = use of an antibody-based method to detect/show a specific


protein in a sample (e.g. skin explant)
• Primary antibody binds to specific antigen (Protein, e.g. Nrf1/MC1R).
• Afterwards dye-labeled secondary antibody binds to primary antibody.

Y Secondary antibody with a dye


Y Primary antibody

Protein (Nrf1 / MC1R)


Fig 2: Schematic representation of the immunostaining method.
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EX VIVO study Results – Nrf2 immunostaining

Negative control Placebo pre-treatment Ectoin® pre-treatment

without visible light irradiation without visible light irradiation without visible light irradiation

with visible light irradiation with visible light irradiation with visible light irradiation
Results: Nrf2

• Nrf2 content in the cell increases automatically, when the skin is exposed
to visible light  Nrf2 protects skin from light-induced oxidative
stress.

• Without visible light / with Ectoin®: Nrf2 content is lower compared to


control  Ectoin® decreases oxidative stress response of the cell. Ectoin®
reduces the need of Nrf2.

• With visible light / with Ectoin®: Nrf2 content is low  Ectoin® reduces
oxidative stress response of the cell. Ectoin® reduces the need of Nrf2.

Ectoin® has been shown to protect skin from visible


light induced skin damage.
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EX VIVO study Results – MC1R immunostaining

Negative control Placebo pre-treatment Ectoin® pre-treatment

without visible light irradiation without visible light irradiation without visible light irradiation

with visible light irradiation with visible light irradiation with visible light irradiation
9
Results: MC1R

• Visible light increases MC1R content and leads to pigmentation of the


skin.

• With visible light / with Ectoin®: reduction of the pigmentation by a


reduction of the light-induced melanogenesis.

Ectoin® has been shown to protect skin from visible


light induced pigmentation.

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Further visible light protection study (in vitro)
• Pre-treatment of keratinocytes with Ectoin© (0.025, 0.05 and 0.1mM).
• Keratinocytes were irradiated by combined light (UVA/visible) and visible light (dose of 15 J/cm).
• Alkaline comet assay.
Visible light (400-800nm wave length)

UVA/visible

Results:
All tested concentrations of Ectoin©
showed highly significant photo
protection on cellular level against the
combination of UVA/visible light (68.9%
protection) and the irradiation with
Control: non-irradiated cells; Irc: untreated irradiated cells.
visible light (92.7% protection).
Source: Botta et al, 2008.
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…get in touch with us

Would you like more information?


Simply contact us. We look forward to hearing from you — and to working with you.

Direct contact with our cosmetics business development:


Phone: +49 163 8603822
E-mail: galik@bitop.de

bitop AG
Stockumer Str. 28
58453 Witten
Germany
Phone: +49 (0) 2302 / 91440-0
Fax: +49 (0) 2302 / 91440-10
Homepage: www.bitop.de
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