38 ANTI-AGEING A 60 *** *** **
*p<0.5 versus initial conditions **p<0.01 versus initial conditions ***p<0.001 versus initial conditions
■ Placebo ■ 0.1% Phytoglycogen B Before
40 *** * 20 *** After 6 weeks
0 1 hour 2 weeks 6 weeks
human foreskin fibroblasts (ATCC, SCRC-1041, USA) were cultured in 75 cm3
Figure 4: Clinical: Hydrating and anti-ageing effects of the phytoglycogen For the quantification of HA and collagen I, tissue culture
flasks in DMEM containing 15% FBS at 37°C and 5% CO2
in DMEM containing reduced glucose (0.2g/L). For the HA measurements, fibroblasts were
grown in 96-well plates and treated or not (control) with 0.025% phytoglycogen for 36 hours. For collagen I evaluation, the fibroblasts were grown on glass coverslips in 12-well plates and treated or not (control) with 0.05% phytoglycogen for 36 hours. All experiments were performed in n=3.
Quantification of intracellular ATP After treatment, the intracellular ATP content of the NHEK was measured using a GloCellTiter- Glo luminescent cell assay (G7571, Promega, USA) according to the manufacturer’s instructions. Briefly, equal volumes assay reagent were added to the cell layers, the plates then placed on an orbital shaker for two minutes to induce cell lysis. After ten minutes stabilization at room
temperature, the luminescent signal was detected on a fluorescent microplate reader (Perkin Elmer). A standard curve was established using rATP (P1132, Promega, USA) and detected ATP was normalised to total protein content of the cells.
HA and collagen I content For the HA quantification, the media supernatant of the fibroblasts was collected after treatment and the amount of HA present in the media measured and quantified using an enzyme-linked immunosorbent assay (ELISA), as per the manufacturer’s instructions. For the collagen production, the cells were washed and fixed with a formalin solution before being processed for immunohistochemistry. Collagen content in the cell was labeled
and visualized with a primary antibody against human type I collagen, and a fluorescent secondary antibody. Cell nuclei were
PERSONAL CARE January 2024 . During treatment, all cells were cultured
Clinical studies To evaluate the clinical efficacy of the phytoglycogen, two double blinded randomized and placebo-controlled studies were performed. In the first study, the hydrating and anti-ageing effects were evaluated on 63 female volunteers aged 51 – 65 years with moderate global facial photodamage and self-perceived dry skin. The volunteers applied a cream containing
0.1% phytoglycogen to the whole face, twice daily, for six weeks. The measured parameters were skin hydration (Corneometer CM 825, Courage + Khazaka, Germany), skin hyperpigmentation (expert clinical grading) and skin even tone (expert clinical grading). Images from select volunteers were acquired
with VISIA CR2 (Canfield Imaging Systems, USA) and the forehead wrinkles, crow’s feet and pigment spots analysed further with Newtone Inc. proprietary technology (Newtone Inc., USA). For the pigment spot analysis, the L*, b*
and a* factors of the skin inside the pigment spot and the skin surrounding the spot were measured an, yielding a difference in contrast (visibility) of the pigment spot. In the second study, the moisturizing
properties of the phytoglycogen was further investigated. Thirteen healthy male and female volunteers aged 25 – 60 years were recruited, and different formulations were tested, containing the phytoglycogen and/or HA (MW 800-1200 kDa, MakingCosmetics, USA). The tested formulations were either a
cream containing 0.1% phytoglycogen, a cream containing 0.1% HA, or a cream containing 0.05% phytoglycogen and 0.05% HA combined. Each volunteer applied all three formulations at defined areas of the inner forearm, a fourth area was added for untreated (no formulation) control.
stained with DAPI. Images were taken using a fluorescent microscope (Nikon Ti Eclipse, Nikon, Japan) and the fluorescence intensity quantified.
After a single application, skin hydration
measurements (Corneometer CM 825, Courage + Khazaka, Germany) were taken over a time course of seven hours and compared to baseline and untreated controls.
Results and discussion The effects of the phytoglycogen on intracellular ATP production was firstly investigated in vitro in keratinocytes. As one of the main energy- carrying molecules found in the cell, ATP drives many important processes, including metabolism, proliferation and the production of essential molecules. After just two hours, treatment with
1% phytoglycogen resulted in a significant stimulation of ATP production (Figure 3A). The levels were further increased after 24 hours by almost 50% compared to untreated controls. Positive effects of the phytoglycogen were
also evident in human fibroblasts. Treatment with 0.025% led to a significant increase in HA secretion from the fibroblasts. Compared to control, HA secretion was increased by 83% (Figure 3B). Furthermore, treatment with 0.05 %
phytoglycogen significantly induced collagen I. In conclusion, the phytoglycogen can induce ATP levels, thereby stimulate cell metabolic activity, which results in the increased production of beneficial molecules such as HA and collagen. The increased HA levels, as observed in the
in vitro assays, can have beneficial effects such as better skin hydration. This was investigated in a clinical study, where the volunteers applied a cream containing 0.1% phytoglycogen over a total of six weeks. Just one hour after the first application, the
treatment significantly increased skin hydration by 53.3% compared to baseline, which demonstrates the immediate efficacy of the plant glycogen. This hydration increase remained
significantly enhanced after two weeks and six weeks of treatment. Furthermore, after six
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compared to initial conditions in %
Change in skin hydration
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