In the meantime, glycation alters the elasticity of the skin through collagen crosslinking and leads to uneven skin tone through the accumulation of advanced glycation end-products (AGE). It is these processes of lipid peroxidation and glycation that trigger a vicious circle of ageing. Plantasens® Olive Active HP is an anti-ageing active ingredient derived from olive that acts as a protective shield. It protects against UV-induced lipid peroxidation by scavenging reactive oxygen species. It also prevents the formation of AGEs and limits collagen crosslinking. It thus helps to maintain the skin’s natural balance and smooth appearance, and its elasticity. Through those mechanisms, Plantasens Olive Active HP aims to sustainably preserve the skin’s natural beauty for longer.

Ageing is a fact of life; but whatever your age, it is important to look and feel at your best.1 While the skin’s natural ageing process cannot be stopped, the right skin care regimen can help slow it down significantly.

The skin is protected naturally by a hydro-lipidic film. However, this is subject to oxidative stress. In a chain reaction, free radicals and reactive oxygen species (ROS) are formed which attack the lipids on the skin’s surface and the cell membrane of both the epidermis and dermis. This impairs the skin’s natural balance and causes the surface to lose its smooth appearance.2

In parallel, oxidation by-products stimulate the glycation process. The interaction of ROS and glucose on the amine group of proteins, after a cascade of reactions, results in the formation of Advanced Glycation End products (AGEs), such as carboxymethyl-lysine – a brownish compound, which gives the skin a dull appearance, and pentosidine – an AGE that fixes onto the collagen fibres, crosslinks them, and forms rigid bridges which make the skin lose its suppleness and resilience.3 Both processes – lipid peroxidation and glycation – trigger a vicious circle of ageing: Oxidation induces glycation and glycation feeds oxidation.

Plantasens Olive Active HP (olive unsaponifiables fraction), sustainable by nature, can help to counteract both of these reactions in the skin ageing process.

Sustainability Aspects

The demand for more sustainable ingredients in personal care products is no longer the exception, but rapidly becoming the norm. A growing number of consumers have a critical eye for what they apply on their skin, and cosmetics manufacturers have turned their attention towards finding effective, sustainable solutions that meet this consumer need.

Olive unsaponifiables fraction is produced from 100% renewable feedstock, using no fossil resources, and has a 100% natural CO2 life cycle. It is also non-food competing as it uses a side stream from olive oil refining. Choosing to use olive origin has no impact on biodiversity as this plant is not endangered and not concerned by biopiracy. On top of that, it is approved as a natural ingredient by natural charters like Ecocert and Cosmos.

This active ingredient fulfills the best-inclass sustainability and performance excellence standards of Clariant’s EcoTain® label; helping customers to realise sustainability targets and gain a unique competitive edge that benefits stakeholders, society and the planet. Olive unsaponifiables fraction helps to ‘sustainably’ slow down the clock on skin ageing.

Results and Discussion

Prevention of Lipid Peroxidation

The anti-ageing effect of Clariant’s olive unsaponifiables fraction starts with protection against free radicals. These belong to the reactive oxygen species (ROS) that play an important role in lipid peroxidation.

A DPPH study was conducted to evaluate the free radical scavenging ability of olive unsaponifiables fraction. DPPH is a stable free radical which in aqueous solution has a distinctive purple colour. When DPPH solution is in contact with compounds with reductive/antioxidant activity, DPPH changes from oxidised state to reductive state, with colour decreasing.

This test method shows that olive unsaponifiables fraction in a concentration of 2.36% can scavenge half of the free radicals (EC50). It highlights its in vitro antioxidant potential.

Ultraviolet (UV) irradiation induces the production of Reactive Oxygen Species that are in part responsible for the genetic and cellular damages (DNA alteration and protein/lipid oxidation) leading to an acceleration of cutaneous ageing. The protective effect of olive unsaponifiables fraction was evaluated in a human skin explant model irradiated by UV, by measuring the ability to decrease UVinduced lipid peroxide formation. The assay was performed using a PeroxiDetect kit. Lipid peroxides oxidise ferrous iron (Fe2+) to ferric iron (Fe3+) which then reacts with xylenol orange to cause a medium coloration observed at 560 nm.

Irradiation of human skin explants with 1.25 J/cm2 of UVB (+18.7 J/cm2 UVA) clearly and significantly induced lipid peroxidation. The treatment of the explants with a positive control, topically applied at 5 mg/cm2, clearly inhibited this effect. These results were expected and validated the assay. Under the experimental conditions of this study, 3% olive unsaponifiables fraction in an oily solvent (caprylic/capric triglycerides), topically applied at 5 mg/cm2, had a significant protective effect (126%) on UV-induced lipid peroxidation in human skin explants (Fig. 1).

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Acting as a protective shield, olive unsaponifiables fraction shows significant antioxidant effects, with free radical scavenging effect and prevention of UV-induced lipid peroxidation.

Prevention of Glycation

In parallel, lipid peroxyl radicals generated during the first oxidation stage have the ability to induce glycation. In order to break down the vicious circle of ageing, it makes sense to check the anti-glycation potential of olive unsaponifiables fraction.

The goal of the first study was to determine in vitro capability of olive unsaponifiables fraction to protect skin dermal fibroblasts against glycation induced by repeated UV exposure. Repeated UV exposure is used as an in vitro cell ageing induction model.

Cells used in the study were human skin fibroblasts and the anti-glycation effect was determined by the AGE dosage by means of ELISA method. The test was carried out in triplicate by using untreated cells and cells treated with olive unsaponifiables fraction at different concentrations. Cells were exposed or not, to three subsequent UV exposures with output energy of UVA+UVB 48 mJ/cm2. Treated cells were incubated for 24 hours with the test substance before the first UV exposure or at least the first measurement.

The results in Figure 2 show that UV exposure activates glycation in the experimental model, increasing significantly AGEs. The treatment of cell culture with olive unsaponifiables fraction at 1% and 5% is capable of significantly and strongly reducing AGE production respectively by 45% and 61%. It can therefore be concluded that olive unsaponifiables fraction shows dose-dependent protecting properties against the production of Advanced Glycation End Products (AGE), highlighting an in vitro anti-glycation effect.

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Anti-glycation activity was further evaluated on human living skin explants, targeting specific skin AGEs: carboxymethyllysine (CML) – a brownish compound which darkens the skin tone; and pentosidine – a substance that crosslinks collagen fibres which makes the skin loose. Glycation activity is determined by the observation of carboxymethyllysine (CML) and pentosidine immunostaining. Skin explants obtained from plastic surgery are treated with methyl glyoxal to induce glycation artificially. Aminoguanidine was used as a positive control to assess the anti-glycation performance and thus validate the test protocol. Olive unsaponifiables fraction at 3% was applied topically on the skin explants every day during seven days, on the basis of 2 mg/cm2. Glycation was induced by the addition of methyl glyoxal to the culture medium on D4, D5 and D7. Histological observation was then made on the dermis of the skin explants, thanks to anti-CML or anti-pentosidine antibodies. Such immunostainings enable microscopical assessment of those specific AGEs.

Figure 3 shows that when glycation is artificially encouraged, an addition of 3% of olive unsaponifiables fraction leads to lower staining intensity than in the nontreated skin, demonstrating significantly fewer traces of CML. Observation made on skin explant exposed to anti-pentosidine antibodies shows similar tendency when the skin was pre-treated with olive unsaponifiables fraction.

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These two studies prove that the active ingredient prevents the formation of advanced glycation end products (AGEs) that damage the skin, and more specifically the CML and pentosidine. It gives opportunities to keep bright skin tone thanks to lower accumulation of CML in the skin and to improve skin elasticity and firmness thanks to lower collagen bridging with pentosidine.

Improvement of Skin Firmness

Finally, the anti-ageing benefits through prevention of UV-induced lipid peroxidation and glycation were evaluated in an in vivo study.

The aim of the study was to assess and to compare the effect on the biomechanical properties of the skin of a test cream versus a placebo cream, applied in halfbody, in normal conditions of use during 56 days, by 21 adults. Recruited volunteers were women aged from 46 to 61 years old who applied creams on the inner forearm twice daily during the two months of the study. Formulation details are described in Table 1, where the only difference between the test cream and the placebo cream is the presence of 5% of olive unsaponifiables fraction. Skin biomechanical properties were recorded at D0, D28 and D56 by Cutometer. Special attention was paid to the extensibility parameters as they reflect the ability of the skin to be stretched. The lower the value, the more the skin is firm and resistant to stretch. In addition, elasticity parameters were examined as they reflect the skin’s resilience (ability to recover its initial stage after stretching). The higher the value, the more the skin is elastic and tonic (Fig. 4).

Records on immediate extensibility (Ue) and maximum extensibility (Uf) for the test cream show a significant and cumulative average reduction of the values over one and two months, respectively –11% and –10% after one month and –19% and –17% after two months. In the same manner, records on the net elasticity (Ur/Ue) and the elastic recovery rate (Ur/Uf) show a significant and progressive increase of the values over one and two months. Net elasticity increases of 11% first and 18% later, when elastic recovery rate only improves after two months by a factor of 14%. This means that olive unsaponifiables fraction positively influences skin resistance to stretch and tonicity over the time.

It is also interesting to observe on Figure 5 that compared to the placebo cream, test cream with 5% olive unsaponifiables fraction improves the resistance to stretch more significantly. Compared to D0, maximum skin extensibility decreases by 17% after two months of application while it is only 13% for the placebo. Performance of the test cream is 30% higher than the placebo in the conditions of the experiment.

This in vivo study shows that olive unsaponifiables fraction significantly improves skin firmness and elasticity after two months of application, with first benefits already measureable after one month. This benefit can be related, at least in part, to the anti-glycation performance of the active on pentosidine and to the lipid peroxidation prevention.

Conclusion

Plantasens Olive Active HP offers a powerful and 100% natural response to the ever-increasing desire to preserve the skin’s natural beauty for longer. This olivederived active ingredient is a unique fraction that consists of 100% pure unsaponifiables. It helps to counteract two important physiological processes found at the root of most consumers’ anti-ageing needs, lipid peroxidation and glycation. It acts as a natural shield on skin and efficiently protects it from UV-induced lipid peroxidation, scavenges Reactive Oxygen Species (ROS), and prevents the formation of Advanced Glycation End products (AGEs). Through the inhibition of carboxymethyllysine synthesis, it should help skin tone to remain bright. Also, after several weeks of application, and through the inhibition of pentosidine, skin elasticity and resistance to stretch is improved. Plantasens Olive Active HP helps to sustainably slow down the clock on skin ageing and as an all-natural ingredient it minimises impact on the environment.

References

1 Grillitsch A, Jenull B. 50+ and the dream of youthful looks [in German]. Journal für Psychologie 2015; 23 (1): 55-79
2 Bickers DR, Athar M. Oxidative stress in the pathogenesis of skin disease. J Invest Dermatol 2006; 126 (12): 2565-75
3 Gkogkolou P, Böhm M. Advanced glycation end products, key players in skin aging?. Dermatoendocrinol 2012; 4 (3): 259-70.