18 ANTI-AGEING
Figure 10: Activity on stress: membrane potential.
DAPI. It is possible to see that both in the case of fibroblasts treated with UV-A and fibroblasts previously treated with Free Q10 and then exposed to UV-A, red- orange pixels appear. These red-orange pixels are due to the fact that when these fibroblasts are exposed to the UVA, their mitochondrial activity increases so their membrane potential (JC1 dye turns red- orange). Conversely, in the case of fibroblasts previously treated with Targeted Q10 and then exposed to UV-A, less re- orange pixels are observed. This means that Targeted Q10 is able to
lower the mitochondrial activity, i.e. the membrane potential, by protecting the fibroblast. It is interesting here to notice that this study has been run at the same concentration of Q10, both in the case of Targeted Q10 and free Q10. We have also measured the fluorescence
and perform a semi-quantitative analysis: Targeted Q10 is able to reduce the membrane potential of mitochondria by 30 per cent. We have also looked at ROS level; Targeted Q10 is able to avoid the production of extra readicals, by lowering radicals level of a 10 per cent (Figure 11). We also studied the ability of Targeted
Q10 to re-energise mitochondria of aged NHDF cells evaluating mitochondrial membrane potential and ROS level. Briefly, NHDF cells have been treated with Targeted Q10 at 0.25% and free Q10 at 0.00005% (this concentration corresponds to the amount of Q10 in Targeted Q10 at 0.25%) for 48 hours. Afterwards, MitoTracker Deep Red immunostaining was performed. Next, cells nuclei have been stained with DAPI (blue color) and mitochondrial membrane potential has been quantified by IN Cell Analyzer device at 633 nm. The total mitochondrial membrane potential increased by 14% after the treatment with Targeted Q10, indicating the product is able to increase the mitochondrial activity, re-energising the mitochondria. Conversely, free Q10 was not able to increase mitochondrial membrane potential. ROS level was also measured. The treatment with Targeted Q10 in
PERSONAL CARE EUROPE
Figure 11: Activity on stress; ROS level.
aged NHDF reduced ROS level by a 5%. This means that even though Targeted Q10 is able to re-energize mitochondria, it is also able to avoid ROS generation and accumulation. Besides the favourable activity of Q10
on mitochondria, and its function as a protector and as an energiser, it has been reported that Q10 is also able to promote the synthesis of ECM proteins, such as collagen type IV and collagen type VII. Collagen type IV is a major component of the basement membrane in the dermal- epidermal junction. Collagen Type IV molecules are thought to act as a scaffold which allows interactions with other non- collagenous basement membrane components, such as laminin, nidogen, and heparan sulfate proteoglycan. It has also been shown that basal keratinocytes preferentially attach to type IV collagen. This interaction apparently contributes to the stability of the skin at the interface between the epidermis and the dermis at the dermal-epidermal basement membrane zone. Also, Collagen Type VII is of particular interest from a dermatologic point of view because it is known to be a predominant component of anchoring fibrils, structures which extend from the dermal-epidermal junction to the upper dermis. This type of structural organisation
of the anchoring fibrils is thought to stabilise the attachment of the dermal- epidermal basement membrane to the underlying dermis. Thus, alterations in the expression, structure, or molecular interactions of type IV and/or type VII procollagens could result in fragility of the skin. For this study, NHDF cells were incubated with Targeted Q10 at 0.025% for 48 hours, and after the incubation, collagen type IV and collagen type VII were quantified by ELISA. The results are shown in Figure 21 A-B. In both cases, an increase of protein production by cells was observed, and in particular Targeted Q10 increased Collagen Type IV and Type VII synthesis by 39% and 27%, respectively.
In vivo study The in vivo efficacy of Targeted Q10 as anti- aging has been evaluated in terms of firmness and elasticity. 18 Volunteers (40-60 years old) were submitted to a 56-day topical treatment with 1.5% Targeted Q10 (0.0003% of Q10) and 1% Free Q10 in the facial area (twice per day, in a hemiface protocol, one treatment in each side of the face). Before (Day 0) and after 56 days of treatment (Day 56), skin firmness and elasticity have been assessed in the crow’s feet area through Cutometer®
statistically analysed. The gathered data are
. All data were
Figure 12: In vivo study: skin firmness and skin elasticity. September 2020
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