ANTI-AGEING 20 15 10 5 0 50% normal fibroblast cells & 50% senescent fibroblast cells treated with Epilobium fleischeri at 0.0005%* ■
Control (50% normal fibroblast cells & 50% senescent fibroblast cells) ■ 100% normal fibroblast cells treated with Epilobium fleischeri at 0.0005%* ■
Control (100% normal fibroblast cells) ■ *:p<0.05 vs control
250 200 150 100 50 0
00:00:00 13:53:20 27:46:40 Time (hours) 41:40:00 * dry extract
Figure 7: AI Analysis of holotomographic images of an in vitro aged skin model showing % dead cells over time, after treatment with Epilobium fleischeri dry extract.
a 94% increase in native collagen I expression (Figure 8).
Senolytic activity on whole tissue Key markers of senescence confirmed that Epilobium fleischeri extract exhibits strong senolytic activity on senescent cells in tissue, as well as in culture. First considering the results for p21, which
is a protein that is essential in inducing cell cycle arrest, as shown in Figure 10, when skin explant tissues from a 60-year-old female were treated with Epilobium fleischeri dry extract at concentrations of 0.005%, 0.024%, and 0.072%, there was a significant reduction in p21 expression. Notably, at 0.024%, Epilobium fleischeri
dry extract decreased p21 by 46%, which was similar to the reduction due to 50 μM navitoclax, a known senolytic drug that inhibits apoptosis. When the SASP marker IL1 was investigated, a 62% reduction was observed in explant tissue treated with 0.072% Epilobium fleischeri dry extract, which was far greater than the reduction due to navitoclax (Figure 9). Also, the expression of Collagen I, which
is a marker for anti-ageing, increased by an impressive 44% in a skin explant treated with 0.024% Epilobium fleischeri dry extract
Senescence marker (p21) 0 -5
-10 -15
-20 -25 -30 -35 -40 -45 -50
0.005
Epilobium fleischeri dry extract (%) 0.024
0.072
Navitoclax (%) 50μM
0
-37 *
-46 *
-46 *
-41 *
-10 -20 -30 -40 -50 -60 -70
0.005
(Figure 9). Taken together, these results confirm that senolytic activity is the basis for the dramatic improvement in explant skin structure following Epilobium fleischeri extract treatment. The micrographs of histologically stained
cross-sections of tissue taken from a 10-day- old untreated skin explant (control) and from a 10-day-old skin explant that had been treated with Epilobium fleischeri dry extract (treated) are shown in Image 3. The Epilobium fleischeri dry extract
treatment was as follows: on the first day, the skin explant from a 60-year-old female volunteer was sampled and treated with various Epilobium fleischeri dry extract concentrations (0.005%, 0.024%, and 0.072%).
Then, on days 3, 5, and 7, the Epilobium
fleischeri dry extract was reapplied, and the skin explants were finally sampled on day 10. Based on the two micrographs shown in Image 3, no toxic effects could be seen after ten days of application. However, the immunostaining of p16 and p21 in the epidermis explant on day 10 confirms that increasing concentrations of Epilobium fleischeri dry extract reduced its intensity such that at 0.072%, Epilobium fleischeri dry extract reduced p16 by 19%
SAAP marker (IL1)
Epilobium fleischeri dry extract (%) 0.024
0.072
Navitoclax (%) 50μM
-17 -30 -40 -62
50 45 40 35 30 25 20 15 10 5 0
194 * 167 * 100
121
0.0005% 0.00025%
Control Epilobium Fleischeri dry extract
Figure 8: Collagen I expression following treatment with Epilobium fleischeri dry extract
(Figure 10) and, at 0.024%, it reduced p21 by 27% (Figure 11), further confirming that Epilobium fleischeri extract can reduce senescent cells, allowing aged skin to regain a more youthful structure.
Clinical study Clinical study is being carried out with subjects applying a cream containing 3% Epilobium fleischeri extract. In one in vivo double- blind study, which ran for three months, two groups of 25 female volunteers with mixed skin ethnicities (skin phototypes II to V) aged between 45 and 60 years (the average age was 55 years) applied a cream twice daily. Their skin elasticity was measured using a
cutometer and photography was used to show fine lines and wrinkles around the eye (e.g., crow’s feet). Skin elasticity and tonicity (Figure 12) had improved by 18% after two months of using the cream with Epilobium fleischeri extract as compared with a control group using a placebo cream. The immediate bounce (R1), which
indicates the skin’s ability to recover after extension, was improved by 18%. The skin’s resilience (i.e., its net elasticity (R5)) had improved by 19%, and an 18% improvement was seen in skin firmness. Using the cream with Epilobium fleischeri
Skin anti-ageing marker (Collagen 1)
44 41
27
3 0.005 0.024 Figure 9: The senolytic and anti-ageing effect of Epilobium fleischeri dry extract treatment on ex vivo 60-year-old female skin
www.personalcaremagazine.com April 2024 PERSONAL CARE 0.072
Epilobium fleischeri dry extract (%)
50μM Navitoclax (%)
Treatment vs control - change (%) Dead (%)
Treatment vs control - change (%)
Treatment vs control - change (%)
Relative collagen 1 expression (%)
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