IBR-CalmDeAge is an extract from palm date (Phoenix dactylifera) seeds. The date seeds extraction preserves the inherent botanical capital of the date seed and delivers it for the most natural active effect on the skin, resulting in a global anti-ageing effect as well as reduction of dark circles and rosacea.
Ancient date seeds that remained dormant for over 2000 years were able to germinate and grow. The dormins preserving the seed youth for such a long period are able to slow down skin cell proliferation, slow down the biological clock and affect intrinsic ageing factors to maintain skin juvenility. The inherent antioxidant and polyphenols equivalent content of the date seed defends the skin against the effects of environmental ageing factors thereby mitigating extrinsic ageing factors. Altogether, this results in a globalised anti-ageing effect. Activation of the alpha-2A adrenergic receptor of smooth muscle cells may result in a vasoconstrictive effect that could lead to improved, homogenous appearance of the skin and reduction of the dark circles and rosacea symptoms. In this article IBR-CalmDeAge’s agonistic effect on alpha-2A adrenergic receptors was observed in addition to a significant anti-irritation effect on pre-irritated tissue. These combined activities are linked to reduction of dark circles and rosacea.
IBR-CalmDeAge is an extract from palm date (Phoenix dactylifera) seeds. IBR’s extraction process preserves the inherent botanical capital of the date seed and delivers it for the most natural effect on the skin. Its effect, observed in vitro and in vivo, includes global slowing down of ageing, mediated by slowdown of cell proliferation and prevention of oxidative damage. Capturing the serenity of the palm’s natural growth environment, IBR-CalmDeAge delivers relief of dark circles and rosacea.
Palm trees are naturally found in oases and are considered a source of life, providing refreshment and rejuvenation, a sign for moisture in the ground, as well as shade which provides protection from the harmful and intense sun irradiation and heat so abundant in the desert. The palm trees are some of the only trees that can tolerate the extreme conditions of the desert. Ancient date seeds were found that remained dormant for over 2000 years. One of these seeds was able to germinate and grow into a viable palm tree. It is thought that the dormins, which kept the seeds viable, but dormant for so long, are also able to slow down skin cell proliferation, thereby preserving skin youth and affecting intrinsic ageing factors.
Slowdown of cell proliferation
According to the Hayflick limit theory of ageing, cells naturally possess a limited number of replications and cell ageing advances with advancement of the replication cycles. It is believed that slowing down cell proliferation preserves cells’ youth capital, thus delaying the ageing process. This groundbreaking theory by Hayflick has formed the basis of substantial research and has recently received strong support with the elucidation of the respective role of telomeres and telomerase as the cells’ internal biological clock. This latest elucidation sheds more light on the cellular ageing process, presenting two possible ways to delay intrinsic ageing: one by slowdown of proliferation and extension of the cell cycle and the other by preventing the shortening of telomeres.
Prevention of oxidative damage
Harman’s free radical theory of ageing suggests that organisms age due to accumulation of free radical damage taking place with the passage of time. A free radical is any molecule that has a single unpaired electron in an outer shell; in most biologically-relevant cases, free radicals are fairly reactive species. For most biological structures, free radical damage is closely associated with oxidative damage. Denham Harman first proposed his Free Radical Theory of Aging in the 1950s with the extension to mitochondrial production of Reactive Oxygen Species (ROS) in the 1970s. Antioxidants are a wide group of substances identified by their ability to capture free radicals and prevent oxidation. This antioxidative ability allows prevention of the subsequent damage that would otherwise contribute to premature ageing.
Rosacea is a chronic condition characterised by facial erythema (redness) as a result of irritation, inflammation and blood capillary imperfections. Rosacea typically presents as redness on the central face across the cheeks, nose, or forehead but can spread and worsen under exposure to the sun, extreme weather conditions, emotional stress and allergens. Rosacea is considered a medically harmless condition, but one that creates aesthetic limitations translated to compromised self-esteem. Rosacea is usually treated with antiinflammatory materials or left not treated at all.
‘Dark circles’ is a term used to identify skin and contour imperfection around the eyes. This term covers the localised puffiness and localised hollowness, as well as shading and skin colour change in the area. Eye puffiness or swelling around the eyes refers to the appearance of swelling in tissues around the eyes (in particular, the lower eyelid), and is usually caused by fluid build-up around the eyes, or periorbital edema. Minor puffiness usually detectable below the eyes only is often called eye bags. Dark circles are brought about by allergies, eczema, eye irritation, anemia, fatigue, heredity, use of vasodilators, bruising, hyper-pigmentation, and skin thinning with age. A common ground for the conditions leading to appearance of dark circles is blood and serum microcirculation, i.e. flow in the capillary vessels under the skin. Be it the result of allergen or autoimmune response, irritation, vascular permeability - blood and serum microcirculation is a common factor. Therefore, improvement of the capillary constriction ability may optimise the flow of fluids in the area. Additionally, reducing skin irritation in combination with the optimised microcirculation could assist in clearing the skin.
Methods and results
In vitro antioxidant effect
• Trolox Equivalent Total Antioxidant Activity (TAA or TEAC) Trolox is a trade name for 6-hydroxy- 2,5,7,8-tetramethylchroman-2-carboxylic acid. Trolox equivalent antioxidant capacity (TEAC), sometimes known as Total Antioxidant Activity (TAA) method is a measurement of antioxidant strength expressed in Trolox Equivalent (TE) units, e.g. mMTrolox. Trolox equivalency is used as a benchmark for the antioxidant capacity of a given substance. Trolox equivalency is measured using the ABTS [2,2’-azino-bis(3- ethylbenzothiazoline-6-sulphonic acid] decolorisation assay, where the antioxidant capacity of a given substance is measured colorimetrically by its ability to decolorise a solution of ABTS• radical (generated from reaction of ABTS with sodium peroxodisulfate), as compared to the decolorising ability of a standard Trolox solution.1
• The Folin-Ciocalteu Reagent (FCR)/The Gallic Acid Equivalence Method (GAE) The Folin-Ciocalteu reagent (FCR) or Folin’s phenol reagent, also referred to as the Gallic Acid Equivalence method (GAE), is a mixture of phosphomolybdate and phosphotungstate used for the colorimetric assay of phenolic and polyphenolic antioxidants. It allows the measurement of the amount of a given substance needed to inhibit the oxidation of the reagent. The sample’s total antioxidant (reducing) activity is compared to that of a gallic acid standard, and expressed as gallic acid equivalent concentrations.2
Total antioxidant activity of 100% IBRCalmDeAge is equivalent to 60-100 mM Trolox. Total polyphenols equivalent of IBRCalmDeAge is: 5000-10000mg/L Gallic Acid Equivalent.
Moderation of cell proliferation
• Dormin-like effect on keratinocyte proliferation Effect of increased concentration of IBRCalmDeAge (now referred to as ‘the palm date extract’) on keratinocyte proliferation, measured using MTT assay for proliferation of Normal Human Epidermal Keratinocytes (NHEK) (Fig. 1).
Anti-irritation: modified Het-Cam irritation test
• Method Irritation reduction in pre-irritated tissue, using a modified Hen’s Egg Test – Chorioallantoic Membrane (HET-CAM) model (n=3). The key measure is the time to appearance of irritation symptoms. A delay is considered as indicating an anti-irritant activity (results in Table 1). The palm date extract at 3% reduced pre-induced irritation by SLS by at least 50%.
Alpha-2A adrenergic receptor, vasoconstriction: effects and suggested mechanism of action
• Alpha-2A adrenergic receptors activation and vasoconstriction The adrenergic receptors are metabotropic (i.e. influencing metabolism) G proteincoupled receptors (GPCR). Many types of cells possess these receptors, and the binding of an agonist (activator) will generally cause a sympathetic response, depending on the receptor being affected (e.g. heart rate modification, vasoconstriction/dilation, fat metabolism changes, etc.). An agonistic effect on the alpha-2A adrenergic receptor of smooth muscle cells may result in a vaso-constrictive effect – meaning the blood vessels in which these cells and receptors are present will constrict and limit the pooling of blood, and lymph in the area, which may result in improved, homogenous appearance of the skin in that area and reduction of the dark circles and rosacea symptoms (Fig. 2).
Test method: cellular dielectric spectroscopy
CDS (Cellular Dielectric Spectroscopy) is a technique that measures impedance changes caused by activation of the target cellular receptor across a cell culture (in this case, of recombinant CHO cells expressing the human alpha-2A receptor). The changes caused by treatment with the test products are compared with the effects of a reference activator (100 nM epinephrine), are expressed as a percentage of the reference’s effect ([measured specific response/control specific agonist response] x 100) Table 2.
Results: cellular dielectric spectroscopy
The palm date extract showed a significant activity as an alpha-2A adrenergic receptor agonist, potentially affecting the capillary blood vessels constriction, and alleviating dark circles and rosacea.
In vivo anti-ageing and anti-rosacea
Assessment of the efficiency, the skin tolerance and the cosmetic qualities
The test was carried out in France on 12 female volunteers aged 50-69. A double blind, randomised hemi-face application, was aimed at assessment of the treatment on skin conditions by image analysis using Visia CR where ageing and rosacea conditions were assessed. The product comprising 5% the palm date extract and its corresponding placebo were applied for 28 days, twice a day. The panel consisted of 12 healthy female adult volunteers aged from 50 to 69 years old, having a variety of skin types and all presenting rosacea. Assessment methods included:
Standardised pictures taken with Visia CR under different light conditions. The pictures were processed with image analysis software in order to evaluate the effect of the product on anti-rosacea and anti-wrinkle effect.
Anti-ageing effect was evaluated by photographing three-quarter left and threequarter right area of the face, under parallel polarised light. The results present percentage of variation obtained after 28 days of use for the length of the wrinkles and the number of wrinkles.
This was evaluated by photographing the face under cross-polarised light (in order to visualise redness) The following parameters were calculated:
• Rosacea surface (average and total) in mm2.• Rosacea density (nb/mm2). • Rosacea average diameter (mm).
Statistical analysis of the parameters measured was performed on the intraindividual differences between D28 and D0. The differences’ significance has been evaluated with a paired Student’s t-test, calculated using a risk of 5%.
Anti-ageing effect (wrinkling)
The cream with the palm date extract shows an average reduction in wrinkles length of 29%. The length and the number of wrinkles decreased after 28 days of use. The product showed a significant antiwrinkle efficiency (length and number of wrinkles decreased after 28 days of use).
Dark circles relief
A reduction in dark circles was observed with the active cream and not with the placebo. These are indicated by arrows on the pictures (Fig. 3).
Rosacea relief effect
The measurements obtained for the volunteers record the variations obtained after 28 days of use for the rosacea parameters (total surface, ratio, and density, average surface and average diameter). A significant decrease of the total surface (–33% average) and of the ratio of rosacea area is presented for the product comprising 5% the palm date extract after 28 days of use by the volunteers (Fig. 4).
The palm date extract is an extract from palm date (Phoenix dactylifera) seeds. IBR’s extraction process preserves the inherent botanical capital of the date seed and delivers it for the most natural effect on the skin. The anti-ageing, as well as reduction of dark circles and rosacea activities of IBR-CalmDeAge, are demonstrated both in vitro and in vivo. The demonstrated antioxidant and dormin-like cell proliferation slowdown effects of the seed’s extract provide support to its global extrinsic and intrinsic anti-aging effect, based on the two key theories of ageing: the Hyflick Limit Theory and Harman’s Free Radical Theory. The in vitro results presented above provide support for the extract’s proposed mechanism of action, via anti-irritant activity and alpha-2 adrenergic receptor agonistic effect which may result in vasoconstriction and optimised microcirculation in the affected area, leading as shown in vivo to reduction of dark circles and relief of rosacea.
1 Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 1999; 26 (9-10): 1231-7. 2 Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folinciocalteu reagent. In: Lester Packer ed. Methods in enzymology Vol 299. San Diego: Academic Press Inc, 1999: 152-78.
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