28 SKIN CARE


120 100 80 60 40 20 0


-45.3% *** (glycation) Control


100 µg/mL Zenerity biotech ingredient concentrate


Figure 2: Percentage inhibition of AGEs formation (*** p < 0.001)


stress. HMGB1 (High Mobility Group Protein B1), one of the most described stressorins in keratinocytes, was measured in the supernatants by ELISA, and the results were normalized with the total viable cell number. As shown in Figure 1, treatment with the


active ingredient reduced HMGB1 stressorin release in vitro, even when induced by metabolic stress, suggesting a reduced inflammatory response. An in tubo glycation reaction was performed


to assess the ability of the active ingredient to counteract AGEs formation. Type I collagen (0.6 mg/mL) was incubated with glucose (0.4 M) and with the active ingredient at 60°C for three days. As a glycation control, only type I collagen


and glucose were incubated. Glycation levels were determined by measuring AGE-specific fluorescence. A decreased formation of AGEs in tubo for the active ingredient was observed, suggesting an anti-inflammatory and anti- glycation effect (Figure 2).


A


250 200 150 100 50 0


20 B


300 250 200 150 100 50 0


Control 30 40 Keratinocyte age (years old)


56 year old cells


-54.0% ** * -50.0%


100 µg/mL Zenerity biotech ingredient concentrate


Metabolic stress


biotech ingredient concentrate


Figure 4: Lipofuscin accumulation. A) Percentage lipofuscin accumulation in HEKa of different donor ages. B) Percentage inhibition of lipofuscin accumulation in HEKa 56 year-old (vs Control: * p < 0.1; vs Metabolic stress: ** p < 0.01). C) Representative confocal images showing the lipofuscin accumulation (red) for different HEKa ages, and treatments


PERSONAL CARE June 2024 www.personalcaremagazine.com


Metabolic stress + 100 µg/mL Zenerity


56 year old cells


100 µg/mL Zenerity biotech ingredient concentrate


50 60


140 120 100 80 60 40 20 0


*** ** -12.4%


-20.0% ****


-20.0% ****


-27.7%


Control


100 µg/mL Zenerity biotech ingredient concentrate


Metabolic stress


15 µg/mL RAGE Antibody


Metabolic stress + 10 µg/mL Zenerity biotech ingredient concentrate


Metabolic stress + 100 µg/mL Zenerity biotech ingredient concentrate


Figure 3: Percentage inhibition of IL1alpha(with symbol) release (vs Control: ** p < 0.01; vs Metabolic stress: *** p < 0.001, **** p < 0.0001).


To further confirm the anti-inflammatory


capacity of the active ingredient, IL1a, a pro- inflammatory cytokine released by RAGE-NF- kB pathway, was measured and compared with the use of a specific RAGE antibody that blocks RAGE activity. HEKa were treated with the active


ingredient with or without metabolic stress for 24 hours. IL1a was measured in the supernatants by alphaLISA, and the results were normalized with the total viable cell number. Treatment with the active ingredient


resulted in a significant reduction of IL1a in vitro, even when induced by metabolic stress, mirroring the effect observed with the use of the RAGE antibody (Figure 3). These findings indicate that the active ingredient can counteract the RAGE-NF-kB activity. Finally, to determine if the active ingredient


could counteract the 'garb-aging' process, HEKa of different donor ages (26, 42 and 56 year-old) were treated with or without


Control ■ Metabolic stress ■ C


26 year old cells


42 year old cells


metabolic stress. Further, the 56 year-old cells were also treated with the active ingredient with or without metabolic stress for 48 hours. Lipofuscin accumulation was evaluated through fluorescence and normalized by the cell number. As shown in Figure 4A and 4C, lipofuscin


accumulation in HEKa exhibited a linear increase with age in HEKa. Across all HEKa age groups, metabolic stress induced an increase in lipofuscin accumulation versus its respective control. Notably, the older the HEKa were, the greater the disparity between basal and metabolic stress-induced lipofuscin accumulation. However, treatment with the active ingredient in older keratinocytes reduced lipofuscin accumulation in vitro, even under metabolic stress, rejuvenating cells to exhibit characteristics similar to younger cells (Figure 4B, 4C). Overall, the biotech ingredient helps


minimize skin inflammation, including when exacerbated by an unhealthy diet which may


Control Metabolic stress


AGE level (%) Lipofuscin accumulation (%) Lipofuscin accumulation (%)


IL1a release (%)


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