132 SKIN PROTECTION
are thus often under protection and cannot be harvested in the wild. Additionally, wild mosses filter the air and retain toxins that prevents them from use for cosmetics. To still make the adaptation skills usable for cosmetics, an innovative biotechnology to grow moss cells in a laboratory setting was developed in close collaboration with Greenovation Biotech GmbH and Prof Dr Reski from the University of Freiburg. Sterile cells of the wild-type moss Physcomitrella patens in liquid culture were produced. Additionally, a new cold pressing extraction method was established to harvest all water soluble ingredients from the moss cells resulting in a natural end product containing potent molecules. The extract was spray dried on an isomalt matrix and the resulting moss active [MossCellTec™ No.1, INCI: Phytol (and) Isomalt (and) Aqua/Water] was investigated for its ability to influence cell nucleus health and skin resilience.
Materials and methods Gene expression in old vs. young keratinocytes Normal human epidermal keratinocytes (NHEK) isolated from a young donor (20 years old female) and an older donor (55 year old female) were cultured for 24 hours. The keratinocytes from the old donor were then incubated or not (control) with different concentrations of Physcomitrella patens extract for 24 hours. All experimental conditions were performed in n=3. Cells were harvested and total RNA was extracted from each sample using TriPure Isolation Reagent (Roche) according to the supplier’s instructions. RT- qPCR for the target genes was performed in n=2 using the LightCycler® (Roche).
system Normal conditions Control 0.33% P. Patens extract 1% P. Patens extract 120 100 80 60 40 20 0 RanBP17 Young cells LAP2B Old cells
Figure 2: Gene expression of cell nucleus health markers in keratinocytes from a young donor and an old donor treated with Physcomitrella patens extract.
Temperature and humidity adaptation of 3D skin model
19 day old 3D human reconstituted skin (Episkin) was treated with 1% moss active or just medium (control). After three hours, climatic stress was induced on the 3D skin either mimicking hot/humid summer stress (40°C, 80% relative humidity, 30 min) or cold/dry winter stress (10°C, 40% relative humidity, 15 min). These climatic stresses were repeated 3 times over 36 hours. Non- climatic stressed 3D skins were included in the experiment as a control. Afterwards, the 3D skins were incubated during 10 hours before processing. After the incubation period, each 3D skin was cryopreserved for histological studies. A Hematoxylin-Eosin staining was performed on one part of the
Hot/humid (40ºC, 80% humidity)
3D skin. On the other part, an immunostaining with an antibody against LCE1A (Thermo Fischer) was performed using a secondary antibody (Alexa 633 anti- Rabbit), together with DAPI staining to visualise cell nuclei. The images were acquired with a Leica confocal microscope. LCE1A levels were quantified with image analysis using the Leica LasX software.
Skin adaptation in a clinical study A double-blind placebo-controlled clinical study on 23 Asian women (39 – 53 years old) having 2-5 hours outdoor activity daily was performed during the summer in Seoul, Korea. The women applied 2% moss active and the corresponding placebo cream on each half of their face twice daily for 14
Cold/dry (10ºC, 40% humidity) Lamin A
Figure 3: Adaptation of 3D skin models treated with the moss active to climatic changes. PERSONAL CARE EUROPE April 2018
1% Moss active
Control
Gene expression compared to young cells (=100) in %
Collapsed dermal structure
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