RENEW EYE SERUM™ Clinical Study XERIDIAN® CLINICAL SKINCARE® General Discussion The Renew Eye Serum™ reverses the development of dark circles and puffiness in the eye area by utilizing acetyl-l-carnitine (a natural skin-energizing amino-peptide), escin (a chestnut seed extract), natural DHEA prohormones, and a broad range of organic botanicals. Dual-stabilized l-ascorbic acid (Vitamin C), centella asiatica extract, green algae extract, and natural beta-glucans work to promote collagen synthesis while natural neuropeptides in this formula provide an immediate wrinkle-relaxing effect within minutes of application. The grape seed extract and resveratrol red wine polyphenols in this product combine with superoxide dismutase and peroxidase (super-antioxidant enzymes) to protect the skin against free radical damage. This active serum can be applied at any time and is especially suggested as an evening-therapy. o Corrects Peri-Orbital Wrinkles – Numerous clinical studies have shown that stabilized forms of LAscorbic Acid (Vitamin C) can dramatically enhance collagen synthesis and act as a potent antioxidant against free radicals. Centella Asiatica, Beta-Glucan (a natural skin-enhancing polysaccharide), Green Algae Extract, and Tara Tree Extract magnify these results by also promoting additional collagen synthesis, protecting existing collagen fibers from degradation, and guarding against free radical damage. o Calms Puffiness & Lightens Dark Circles – The high concentration of Acetyl-L-Carnitine (a natural skinenergizing amino-peptide), Escin (extracted from natural chestnut seeds), GHK Tripeptides (natural anti-aging amino-peptides), Butchersbroom Extract, Ivy Extract, and Natural DHEA Prohormones take an active role in aiding in the repair of vascular walls to discourage the pooling of iron and hemoglobin that cumulatively appear as dark circles. This not only promotes proper microcirculation but also encourages the draining of excess fluids and swelling that contribute to puffiness. o Antioxidant Protection For Delicate Undereye Skin – In addition to the potent antioxidant activity of Vitamin C, this formula also includes Grape Seed Extract and Resveratrol (a red wine polyphenol) along with Superoxide Dismutase and Peroxidase super-antioxidant enzymes to protect the skin against free radical damage. o Skin Cell Life Extending – The Resveratrol in this formula is a grape polyphenol that gives red wine much of its antioxidant attributes. Laboratory studies have shown that this compound can greatly extend the lifespan of cells by mimicking the beneficial aspects of caloric restriction and new research is constantly coming out pointing to the implications this has on extending youthful skin cell life. Formula Active Mechanisms Anti-Wrinkle Ingredients L-Ascorbic acid is a water-soluble form of vitamin C chirally shifted left to be more effectively incorporated into the skin, where it can act not only as an essential antioxidant, but also has been shown in clinical studies to stimulate collagen synthesis (which produces visibly tighter and smoother skin over time) as well as lighten hyperpigmentation. It is also a natural free radical scavenger that works synergistically with other antioxidants in the skin. Because pure ascorbic acid is very unstable in aqueous formulas we use dual-stabilized forms of Vitamin C that clinical studies have demonstrated have long-term stability when used in topical products. Recently a bright spotlight has been on the extract of Centella Asiatica (a herb native to India) as having powerful antioxidant properties, and clinical tests have demonstrated that it can penetrate deeply into the skin to stimulate collagen synthesis (which in turn promotes skin tightening and a reduction of wrinkle depth). Beta-Glucan is a purified extract from natural Oat Straw that provides many benefits to the skin, including increased hydration, increased skin firmness, and an increase in skin elasticity. This specially purified form of green algae contains marine pentasaccharides (MPS) that readily penetrate the epidermis to promote the synthesis of collagen and function as metabolism mediators within the dermis. In the dermal layer these MPS compounds will function to inhibit the synthesis of Metalloproteinases (MMP) as well as the synthesis of pro-inflammatory mediators such as cytokines. When MMP is inhibited, the degradation of collagen and other extracellular components is reduced. The result is less wrinkled, rejuvenated, and firmer skin. Stabilized Vitamin C Centella Asiatica Extract Beta-Glucan Green Algae Extract Undereye Clarifying Compounds Escin Escin is the active constituent of Horse Chestnut Seed Extract that has been used in undereye formulas for decades as one of the most tested ingredients for the repair of vascular walls. Because of these venotonic and anti-edematous properties it is able to keep capillaries from leaking fluids that show through the thin skin under the eyes as dark circles and puffiness. This extract contains an abundance of phytonutrients that tighten the skin of the undereye area and regulate blood flow in capillaries. Most notably the caffeine content of this extract has both immediate and long-term effects with daily use in diminishing both dark circles and puffiness due to swelling. This ingredient also has the added benefit of tackling puffiness through reducing the volume of fatty tissue under the eyes. A common surgery for baggy eyes is removal of fatty tissues (a procedure known as a lower blephorplasty) that contribute to puffiness. This extract causes the individual fat cells to constrict, giving the undereye region both a smoother and tighter appearance. Ivy extract contains the phyto-compound Hederine, which is an active saponin responsible for blood vessel protection. It specifically reduces the permeability of capillaries to keep fluids from accumulating in the undereye region. Ivy Extract also helps reabsorb the edemas present in fatty tissues under the eyes. The botanical extract of Burchersbroom acts mainly on microcirculation in the undereye area, decreasing capillary permeability due to its content of a flavonoid called Rutin. DHEA is a natural prohormone produced by the body as a precursor to other hormones and also has direct anti-aging effects itself. As we age DHEA production decreases and this causes levels of other critical hormones to which it is a precursor to also decrease. Ultimately, this results in thinner skin that is more easily damaged and less elastic. Several studies have shown that DHEA is easily absorbed topically and has direct implications for skin thickening, which is critical not only for elasticity around the eye area but also decreases dark circles by providing a more opaque barrier between the skin’s visible surface and the underlying capillary network. The term “neuropeptides” describes specific peptide compounds that have been shown to have an effect on neurotransmitters in the skin (one that typically translates to a “relaxing” effect on wrinkles). While only wrinkle-relaxing injections can actually paralyze the underlying muscles of the face that affect facial expressions, there are actually subtle tensing contractions that go on in the skin itself. Often referred to as “micro-tension,” this is a more subtle yet still noticeable affect of contractions in the skin that show through as fine lines and deeper wrinkles. Antioxidant Compounds Grape Seed Extract Grape Seed Extract contains antioxidants that studies have proven to be 50-times more effective than vitamin E and 20-times more effective than vitamin C in scavenging for free radicals. It also has soothing and reparative properties through its ability to strengthen capillaries, reduce inflammation, and strengthen skin’s defenses against UVB radiation. Resveratrol is a recently discovered polyphenol found in particularly high concentrations in the skins of red grapes and is often attributed to the health benefits of moderate red wine consumption. It is a potent antioxidant that has a particularly beneficial affinity for neutralizing the free radicals most responsible for lipid oxidation. This is thought to be the mechanism that gives red wine its antioxidant and heart health promoting attributes. Recent studies in the laboratory have shown that Resveratrol can greatly extend the lifespan of cell cultures, and while further studies are needed to confirm these results in humans there is strong indication they can have life-extending effects on the youthful functioning of skin cells. Superoxide Dismutase (SOD) is a powerful enzyme complex that helps to counteract the damaging effects of various reactive oxygen species (ROS) and is therefore an extremely useful antioxidant in the skin. Superoxide Dismutase neutralizes the superoxide free radical anion and numerous clinical studies have demonstrated that topical application of SOD serves to reduce irritation/inflammation in addition to its antioxidant benefits. Superoxide dismutase in conjunction with peroxidase is responsible for the scavenging and neutralization of both the Superoxide and Peroxide radicals which contribute to premature aging. They form a powerful duo together because when Superoxide Dismutase neutralizes the superoxide anion it can release peroxide, which is subsequently neutralized by Peroxidase. Coffea Arabiga Seed Extract Ivy Extract Butchersbroom Extract DHEA Prohormones Neuropeptides Resveratrol Superoxide Dismutase Peroxidase Figure 1: Study Participant Age Range Figure 2: Study Participant Skin Type Range Figure 3: Fitzpatrick Skin Type Range There were a total of 16 participants in this study, composed of 11 females and 5 males. The age of these participants ranged from 30 to 71 with a median age of 51 (Figure 1). The participants were asked to stop the use of all other skincare products they may be using for one month prior to the study as well as during the study (except for products that were not “leave-on” such as cleansing or shower products). All participants were given the RENEW EYE SERUM to use twice daily for a ten week period. 7% 0% 20% During the initial intake participants were asked to complete a self-evaluation questionnaire in which they assessed various characteristics of their own skin. Most notably this included a self-evaluation of their basic skin type: oily (excess sebum production resulting in greasy skin and/or glossy shine), combination (oily T-zone around the nose and/or forehead but dry skin elsewhere on the face), normal (hydrated but not oily skin), and dry (dull, flakey, and/or dehydrated skin that does not have enough sebum oil production to keep the skin moist). During the participant intake the tonality of the skin was assessed using a Fitzpatrick Skin Type Scale with one (I) representing very light pigmented skin, two (II) representing medium-light pigmented skin, three (III) representing medium skin, four (IV) representing medium-dark skin pigmentation, and (V) representing dark skin pigmentation. No one in this particular study had a Fitzpatrick Skin Type of VI, which is very dark pigmentation. 13% 6% 43% 19% 6% 32% 27% 25% 20% 26% 25% < 29 50‐59 30‐39 60‐69 40‐49 >70 Oily Normal Combination Dry I II III 31% IV V Study Methodology & Results To determine an absolute measurement of wrinkle depth a standard Skin-Visiometer® SV 600 was used, which evaluates the topography of the skin surface by light transmission of a thin blue dye silicone. This specially prepared silicone, mixed under a vacuum to avoid bubble formation, permeates even the smallest skin depths to form a micro-cast that is evaluated by a parallel light source and CMOS camera to quantitatively measure the heights and depths of the skin surface area as a negative. The statistical mean results of the study participants are presented in the graph below. 60% 50% 40% 30% 20% 10% 0% For the purposes of measuring dark circle coloration in the skin a standard Mexameter® MX 18 was used. The probe emits three specific wavelengths of light reflected by the skin in a receiver and the quantity of light absorbed by the skin can be calculated. To assess undereye skin discoloration skin using haemoglobin pigment as an indicator, specific wavelengths of light are used corresponding to the spectral absorption of peak haemoglobin at baseline then compared to final saturation at study end. The statistical mean results of the study participants are presented in the graph below. 51% 40% 30% 20% 10% 0% 38% 29% 35% 39% 27% 0% 0% Day 0 (Baseline) Day 35 Day 70 Average Percentage Improvement In Deep Wrinkle Depth As Measured With Skin‐Visiometer® SV 600 Average Percentage Improvement In Fine Line Depth As Measured With Skin‐Visiometer® SV 600 0% Day 0 (Baseline) Day 35 Day 70 Average Percentage Improvement In Dark Circles As Measured With Mexameter® MX 18 BEFORE: This study participant has moderately deep wrinkles that extend laterally to the sides of the eyes as well as fine lines. She also has moderate dark circle formation under the eyes along with some hyperpigmentation throughout the periorbital skin region. The skin also has some moderate redness from underlying irritation. AFTER: After ten weeks the deep wrinkles that extended laterally from the sides of the eyes have greatly diminished and the fine lines surrounding the peri-orbital region have softened considerably. The hyperpigmentation that dotted the skin around the eyes has lightened tremendously. BEFORE: This study participant has moderate dark circle discoloration under his eyes as well as some underlying skin redness/inflammation. The fine lines extending out from the peri-orbital region are pronounced and will give way to more permanent deeper wrinkles (“crow’s feet”) if not corrected. Some puffiness in the undereye skin is also noticeable. AFTER: After ten weeks of RENEW EYE SERUM use this study participant has dramatically lessened his undereye darkness and largely resolved the appearance of discoloration. The fine lines that were previously very apparent have diminished to the point of being almost invisible. The puffiness under his eyes has significantly decreased giving the skin a smoother and more uniform appearance. BEFORE: This study participant has fine lines primarily under the eyes that parallel the eyelids. A minor amount of dark circle development is also noticeable as well as light hyperpigmentation. AFTER: After ten weeks the fine lines surrounding her eyes have greatly diminished to the point of being barely noticeable. The darkness that was beginning to form under her eyes has completely faded and the hyperpigmentation spots have lightened considerably. BEFORE: This is the same study participant as above shown at an angle to more fully capture the extent of fine line development under her eyes that was less apparent in the straight on photos. In addition to several fine lines surrounding the undereye area there also is noticeable puffiness that is more visible from this profile angle. AFTER: The fine lines have dramatically decreased giving a much more youthful appearance to the eye area. The puffiness under the eyes has largely subsided and the minor amount of hyperpigmentation that was present is not considerably more evened out to the rest of the skin tone.