56 EYE AREA CARE


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Placebo


3.0% HB Complex


Figure 6: HB Complex leads to a stronger reduction of the surface area of dark circles than placebo.


Results Human epidermal equivalents with melanocytes, when exposed to haeme, clearly show an increased production of melanin. In this setting, the treatment with HB Complex led to a strong reduction of melanin production (Fig 3). Exposure of these epidermal skin equivalents to both haeme and UV radiation shows a further increase in melanin production. Here, the treatment with HB Complex shows a greater reduction of melanin production than with the exposure to haeme only. These results clearly demonstrate that HB Complex reduces melanin production under circumstances which are extremely relevant to the formation and persistence of dark circles.


VEGF-C and lymphatic endothelial cells VEGF-C (Vascular Endothelial Growth Factor-C) is a growth factor which promotes the production of functional lymphatic vessels. These are of eminent importance in the aetiology of dark circles, as described above. Inducing the production of VEGF-C is therefore extremely important in fighting dark circles and puffy eyes. In the eye area, fluid can accumulate relatively easily. This fluid contains cells, ions and macromolecules (specifically proteins). The ions lead to osmotic pressure, whereas the proteins cause a phenomenon called oncotic pressure. In an experiment that determines the effect of an active ingredient of VEGF-C expression, osmotic pressure should therefore be taken into account. As keratinocytes play an essential role in cellular communication, including toward the lymphatic endothelial cells, it is of particular interest to analyse the effect of HB Complex in an experimental design where the effect of keratinocyte communication to lymphatic endothelial cells, relevant to VEGF-C expression, is determined.


Method Human keratinocytes (HaCaT) were pre-treated with HB Complex for 24 hours. Osmolarity of the medium was adjusted by NaCl solution, and cells were incubated for a further 24 hours. The medium was transferred to the human dermal lymphatic endothelial cells (HDLEC), which were further incubated for 24 hours. The determination of VEGF-C was performed


PERSONAL CARE March 2021


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Placebo


3.0% HB Complex


Figure 7: HB Complex leads to a stronger reduction of the colour of dark circles than placebo.


by ELISA (R&D Systems, Inc.; #DVEC00). Absorbance was measured at 450 nm/540 nm. Non-treated, non-damaged cells served as controls and were set at 0%.


Results Osmotic pressure is stressful for keratinocytes. The cultivation of lymphatic endothelial cells in a medium which originates from hyperosmotically stressed keratinocytes led to a slight increase in VEGF-C (Fig 4). In an identical setting that also included the treatment of the keratinocytes by HB Complex, a much clearer increase in VEGF-C production by lymphatic endothelial cells could be observed. Cultivation of the lymphatic endothelial


vessels in a medium originating from the treatment of keratinocytes at normal osmolarity and in the presence of HB Complex also increased VEGF-C production, albeit slightly. These results indicate that HB Complex supports the formation of new and functional lymph vessels. HB Complex thus plays an important role in the promotion of drainage of excess fluid, inflammatory cells and haeme from the interstitial area in the dermis, an activity which is essential in fighting dark circles.


Efficacy studies – in vivo assays Reduction of dark circles In order to assess whether HB Complex reduces dark circles, a study was performed where 12 volunteers applied cosmetic formulations, one containing 3% HB Complex and a corresponding placebo, for 56 days, twice daily on each eye area. At the beginning of the study and after 56 days of application, standardised photographs were taken with a VISIA® device (Canfield Scientific, Inc.). This device allows for detailed imaging in a standardised environment, preventing external light influences and including comparison of different pictures of the same person taken at different times. The standardised photographs were then


digitally analysed by Newtone Technologies (Lyon, France). By making use of the combined technologies offered by Canfield and Newtone, conclusions can be drawn on the effect of a cosmetic product on dark circles. Through the digital analysis of the


photographs, important parameters relevant to dark circles are generated, for instance the


ITA° (Individual Typological Angle). The ITA° is a relevant parameter for darkness of skin. Overall, a reduction of dark circles is obtained when ITA° increases in value. An essential feature in the reduction of dark


circles is their surface area, which is extraordinarily important for decreasing their visibility.


Results Individual results show that the application of 3% HB Complex for 56 days clearly led to a reduction of the surface area of dark circles (Fig 5). An analysis of the effects obtained with 3%


HB Complex and a placebo formulation, now with a focus on the actual surface area of the dark circles on all individuals (Fig 6), clearly shows that 3% HB Complex reduces the surface area of the dark circles more effectively than placebo. To further underline the effects of HB


Complex on dark circles, an additional analysis was performed on the parameters relevant to their colour and visibility. The application of a formulation containing 3% HB Complex led to an increase in value of the parameter ITA° that was clearly larger than that obtained with placebo (Fig 7).


Conclusion HB Complex is a product based on extracts of Bellis perennis and Hieracium pilosella (common names: daisy flower and hawkweed) respectively. HB Complex addresses the complex issues of under-eye dark circles in multiple essential ways. It supports the processes of lymph drainage and reduces the presence of darkly coloured and strongly inflammatory haeme. It also reduces melanin deposition. With these actions it eliminates the presence of pigments which play an important role in the colour intensity of dark circles and takes away important triggers in their pathophysiology. Additionally, it improves cellular functionality and health in skin cells. Thus HB Complex provides essential support in reducing the likelihood of the formation of dark circles.


The activity of HB Complex on dark circles was


proven in two separate in vivo studies against a placebo, i.e. vehicle control. It reduces both the surface area and the intensity of dark circles. HB Complex is a natural, potent and safe active ingredient for skin care for the eye area.


PC www.personalcaremagazine.com


Reduction in surface area of dark circles (%)


Increase in ITA° (%)


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