66 TRENDING TECHNOLOGIES These observations suggest a possible direct
effect of the polyphenols on stimulating and normalizing the function of the Krebs cycle and energy production. The polyphenols could also resolve the impairment of the mitochondrial respiratory chain induced by the pollutants.
Reducing mitophagy The autophagy process is an intracellular recycling pathway that leads to the degradation of cytoplasmic components (malformed proteins, damaged compounds or organelles) within lysosomes. Mitophagy is a specialized type of autophagy which leads to the degradation of mitochondria.14, 15 Figures 9 and 10 show holy basil
polyphenols reduced and normalized pollutant stress-induced mitophagy. According to these results, on one hand, holy basil polyphenols are able to restore mitochondrial function, and on the other hand, modulate mitophagy.
Anti-inflammatory activity Inflammatory response is part of inflammaging. This body’s normal, immediate and transient response to any environmental aggression can lead to tissue damage under certain pathological or physiological conditions. Figure 11 shows holy basil polyphenols
significantly inhibited the production of PMA- induced IL8 (an inflammatory cytokine) in keratinocytes.
Stimulation dermal matrix The dermal matrix is modified with skin ageing, especially with a decrease in the quality and quantity of matrix fibres such as collagen. The activity of holy basil polyphenols on the expression and production of dermal matrix markers was evaluated on normal human dermal fibroblasts in culture. Holy basil polyphenols significantly
stimulated collagen I labeling in normal human dermal fibroblasts. Figure 12 shows the potential of the polyphenols on densification of the dermal matrix and for anti-ageing efficacy.
IL8 PRODUCTION
8000.000 7000.000 6000.000 5000.000 4000.000 3000.000 2000.000 1000.000 0.00
$$$
-40% **
-36% Is
-50% *
-41% *
0.035 0.03
0.025 0.02 0.015 0.01
0.005 0
$$$ * *** MITOPHAGY
40 35 30 25 20 15 10 5 0
$$$
*** ns*** vs Ctrle
Figures 9 & 10: Normal human dermal fibroblasts were incubated for 120 minutes in the presence of a mixture of 0.1% urban pollutants and the polyphenols of holy basil (HB) at 0.001% and 0.005% dm. The co-location of the damaged mitochondria in the lysosomes (dual Red/Green labeling) makes it possible to evaluate the mitophagy process. After image analysis, the percentage of mitochondria damaged per cell, lysosome density per cell (lysosomal index representative of autophagy), the percentage of lysosomes including damaged mitochondria (mitophagic index representative of mitophagy) were monitored
Conclusion of in vitro studies Holy basil polyphenols help preserve mitochondrial function against stress caused by pollution, they provide antioxidant protection for skin, including against psychological stress while modulating inflammation and preserving the integrity of the dermal matrix. Thus, holy basil polyphenols have a significant potential against inflammaging.
Clinical studies on holy basil polyphenols Protocol The study was carried out in a double-blind, comparative (active vs placebo) manner. The subjects applied the cream containing the active ingredient or the placebo (randomization of subjects at inclusion) for 56 days, twice a day (morning and evening). The inclusion criteria defined for the
recruited subjects were: ■ Sex: Female. ■ Age: between 40 and 60 years.
COLLAGEN I PRODUCTION
3000 2500 2000 1500 1000 500 0
Figure 11: IL8 production by keratinocytes under PMA-induced inflammatory stress. Pre- incubation for 24 hours with holy basil (HB) polyphenols at 0.001%, 0.005% or 0.01% dm or 0.1 μM dexamethasone, an anti-inflammatory reference. Quantification by ELISA assay. $$$ p<0.001 vs. Untreated Control; ls p<0.1; *p<0.05; ***p<0.01 vs. PMA - Student t test.
PERSONAL CARE June 2022 +20% +13% +19%
■ Phototype: I to III. ■ Type: Caucasian. Subject with wrinkles: crow’s feet (score
between 3 and 5 on the Bazin scale); nasolabial fold (score between 2 and 5 on the Bazin scale); lips (score between 1 and 4 on the Bazin scale); forehead (score between 2 and 4 on the Bazin scale). Stop all cosmetic product application
between D-6 and D0. The group that applied the active
ingredient consisted of 22 subjects aged 53.9 ± 4.3 years (minimum: 46, maximum: 60). Meanwhile, the group that applied the placebo consisted of 23 subjects aged 52.2 ± 5.3 years (minimum: 43, maximum: 60). Efficacy was evaluated using the following
techniques: ■ Acquisition of a DermaTop 3D image followed by analysis by Newtone Technologies. ■ Scoring by a trained panelist at each time point. ■ Measurement of skin biomechanical properties using the Cutometer. ■ Cross-polarized light photographs using Colorface, followed by analysis by Newtone Technologies. ■ Illustrative photographs in normal light using Colorface.
Figure 12: Quantification of collagen labeling. Normal human dermal fibroblasts were treated for 48 hours with holy basil (HB) polyphenols at 0.001% and 0.005% dm or with a 10 µM Vitamin C and 5 ng/ml TGFβ mixture, used as a positive reference. Collagen I production was evaluated by immunofluorescence labeling and image analysis
Study procedure Subjects came to the clinical unit on D0. Subjects were randomized. Basal measurements were then taken. The products (active ingredient and placebo) were applied by the subjects themselves in their home, twice a day (morning and evening) for 56 days from D0 to D56 on the entire face with an emphasis on the various measurement areas (wrinkles, temple, cheek, etc.). After 28 days, the subjects returned to the
clinical unit for the intermediate time point. The D28 measurements were then carried out. After 56 days, the subjects returned to the clinical unit for the final time point. The D56 measurements were then carried out. The process is summarized in Figures 13 & 14.
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IL8 (pg/ml)
Collagen I (Arbitrary unit)
Lysosome density (intensity/µm2
)
% of damaged mitochondria found in Iysosme
Control
Polluant BS 0.001% ms BS 0.005% ms
Control Pollutant HB 0.001% ms HB 0.005% ms
Control
PMA 10µg/ml Dexa 0.1µM HB 0.001% ms HB 0.005% ms HB 0.01% ms
Control
Vit C 10µM + TGFb 5ng/mL
HB 0.001% ms HB 0.005% ms
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