ANTI-POLLUTION Dermal Fibroblasts
100 80 60 40 20 0
Untreated Cadmium *** Cadmium +
Citrus limon fruit extract active
Figure 8: Quantification of cell viability in dermal fibroblasts after exposure to cadmium heavy metal and treatment with citrus limon fruit extract active during 24h
the activity some protein. This protein has very beneficial activities that guarantee cell homeostasis and genomic stability that promotes longevity.2
In addition, Sirtuin 6 has already
demonstrated its protective effect on cells against the harmful effects of cigarette smoke.3
In vitro tests: protein quantification in cells (Cells in basal conditions) NHDF were seeded in 96 wells plate for 24 hours. Then NHDF were incubated with 10ppm of the citrus limon fruit extract active molecule. After 24 hours, cells were washed and fixed with formalin. After permeabilization, cells were incubated with anti-Sirtuin 6 antibodies and revealed with a secondary antibody linked to fluorescein. Finally, cells nuclei were marked with DAPI. The labelled protein was observed and quantified by automated fluorescence microscopy (ArrayscanTM
Cellomics). In basal conditions, citrus limon fruit extract
active molecule activates the expression of the protein Sirtuin 6 by 14% in the nuclei of dermal fibroblasts (Figure 7). This activation is consistent with the increase in SIRT6 gene expression
DAY 0
110 100 90 80 70 60 50
Untreated
Cigarette extra
Dermal Fibroblasts ***
140 120 100 80 60 40 20 0
Cadmium + Citrus limon fruit extract active 10ppm
Untreated
Cigarette extra
Keratinocytes ***
45
Cadmium + Citrus limon fruit extract active 10ppm
Figure 9: Quantification of cell viability in dermal fibroblasts and keratinocytes culture after exposure to cigarette smoke extract and treatment with citrus limon fruit extract active during 24h
UNTREATED UVA
UVA + CITRUS LIMON FRUIT EXTRACT ACTIVE
Figure 10: Quantification of Lamin B1 labelled by immunofluorescence in skin explants after exposure UVA and treatment with citrus limon fruit extract active (2%)
observed previously. Thus, Sirtuin 6 will bring its beneficial effects on cell homeostasis and genomic stability within cells and particularly on cells exposed to pollution stress.3
In vitro tests: quantification of cell viability under pollution stress NHDF and NHEK were seeded and exposed to the heavy metal cadmium or a cigarette
DAY 28
smoke extract in the presence of 10ppm of citrus limon fruit extract active molecule. After 24 hours, cell viability was measured using the WTS-1 reagent (Roche), this reagent contains tetrazolium salts, a purple indicator. It is cleaved into formazan, a yellow indicator, by metabolically active cells. The level of yellow coloration is therefore proportional to the number of living cells. Absorbance is measured at 450nm. Regardless of the stress factor applied, citrus limon fruit extract active maintains the cell viability the cellular viability of skin cells exposed to polluting materials. Indeed, the viability of fibroblasts decreases in the presence of cadmium (-25%) and is maintained close to 100% in the presence of citrus limon fruit extract active (Figure 8). Similarly, viability decreases in cells
exposed to cigarette smoke extract, -43% in keratinocytes and -15% in fibroblasts, while no loss of viability is observed in the presence of the citrus limon fruit extract active (Figure 9). Thus, the citrus limon fruit extract has made it possible to adapt cellular metabolism in the presence of pollutants in order to preserve the longevity of skin cells.
Figure 11: Example of skin roughness reduction observed in a volunteer who used a cream containing 1% citrus limon fruit extract active for 28 days
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Ex vivo tests: protein quantification in skin explants exposed to UVA (Lamin B1) Lamin B1 is a nuclear protein which establishes a two-dimensional matrix in the internal nuclear membrane. It is involved in the nuclear stability, the structuration of the chromatin and the gene expression control.
July 2024 PERSONAL CARE
% Cell viability
% Cell viability
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