International Journal of Cosmetic Science, 2012, 34, 416423

doi: 10.1111/j.1468-2494.2012.00731.x

Review Article

Interaction of mineral salts with the skin: a literature survey

T. G. Polefka*, R. J. Bianchini and S. Shapiro
*Life Science Solutions, LLC, Somerset NJ, 08873, Merck Consumer Care, Summit NJ, 07901 and Skinnovations, LLC, Roseland NJ, 07068, USA

Received 19 January 2012, Accepted 29 May 2012

Keywords: copper, essential minerals, magnesium, nutrition, skin, zinc

Synopsis There is growing scientic evidence that the health, well-being and the attractiveness of the skin are strongly inuenced by nutrition. Consumers recognize this and have supported the creation of a global cosmeceuticals market estimated in 2010 at $27.2 billion. Early in 2011, the US Department of Health and Human Services and Department of Agriculture issued the Dietary Guidelines for Americans, 2010. Twelve vitamins and nine minerals were recognized as essential. The minerals include calcium, copper, iron, magnesium, phosphorus, selenium, zinc, potassium and sodium. Although the topical benets of several minerals such as zinc, magnesium and iron are recognized and, in some cases, approved by the FDA, the topical benets of the others to the skin are largely unexplored and unexploited. This review attempts to summarize what has been published in the literature on the interactions of the eight of the nine essential elements with the skin. sume Re Les preuves saccumulent pour indiquer que la nutrition inuence , le bien-e tre et lattractivite de la peau. Les confortement la sante tablir un marche sommateurs en sont conscients et ont permis de ments alimentaires nutracosme tiques estime mondial de comple ` 27,2 milliards de dollars. De ` res de but 2011, les Ministe en 2010 a et de lAgriculture des USA ont e dite un guide die te tique la Sante ` la population ame a ricaine. Ce guide reconna adresse t douze vita raux en tant que le ments essentiels. Les mine rmines et neuf mine sium, le aux incluent le calcium, le cuivre, le fer, le magne le nium, le zinc, le potassium et le sodium. Bien que phosphore, le se s des mine raux tels que le zinc, le magne sium et les bienfaits cutane s par la FDA, les be ne ces le fer sont reconnus et parfois approuve ` ce jour, largement sousau niveau de la peau des autres sont, a s et sous-exploite s. La pre sente revue se propose de re sumer explore te publie dans la litte rature sur les interactions entre les ce qui a e le ments mine raux essentiels avec la peau. huit (sur neuf) e Introduction Over the past several years, consumers have begun to recognize and appreciate the role proper nutrition plays in the health, well-being and attractiveness of their skin. Responding to this
Correspondence: Thomas G. Polefka, Life Science Solutions, LLC, 79 Ellison Rd, Somerset, NJ 08873-2257, USA. Tel.: +1 732 221 6889; e-mail: tpolefka@lifescisolutions.com

trend, the cosmetic industry has begun to offer the consumer topical products better known as cosmeceuticals to topically nourish their hair and skin. In a 2010 report, RNCOS estimated the global cosmeceuticals market at $27.2 billion [1]. In early 2011, the US Departments of Agriculture and Health and Human Services issued their 7th edition of the Dietary Guidelines for Americans, 2010 [2]. Accordingly, twelve vitamins (e.g. vitamin A, vitamin D, vitamin C, vitamin E, vitamin K, thiamin, riboavin, niacin, vitamin B6, folate, vitamin B12 and choline) and nine minerals (e.g. calcium, iron, magnesium, phosphorus, sodium, zinc, copper and selenium) are recognized as necessary for health and well-being. Table I lists the nine minerals and their dailyrecommended intakes [2]. Interestingly, Voet and Voet [3] estimate that nearly one-third of all known enzymes require the presence of metal ions for catalytic activity. Table II lists several examples for each mineral. However, discussions on the benets of dietary vitamin and mineral supplementation almost always generate a debate on what level is needed to prevent deciency or disease and what level is required for optimum health and/or well-being. The latter is much more difcult to demonstrate scientically. In contrast to other cosmetic ingredients such as vitamins [4] and botanicals [5], the benets of topically applied mineral salts have been largely ignored and unexploited. Indeed, to the best of our knowledge, few, if any clinical studies
Table I Mineral goals for adults based on dietary reference intakes and dietary guideline recommendations

Mineral

Female 1950 year

Male 1950 year

Female 51 + year

Male 51 + year

Calcium (mg) Iron (mg) Magnesium (mg) Phosphorus (mg) Potassium (mg) Sodium (mg) Zinc (mg) Copper (lg) Selenium (lg)

RDA* RDA RDA RDA AI UL RDA RDA RDA

1000 18 310320 700 4700 <2300 8 900 55

1000 8 400420 700 4700 <2300 11 900 55

1200 8 320 700 4700 <2300 8 900 55

1200 8 420 700 4700 <2300 11 900 55

*Recommended dietary allowance. Adequate intake. Upper limit. Adapted from reference [2].

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Interaction of Mineral Salts with the Skin

Table II Examples of Metalloenzymes

T. G. Polefka et al.

Macronutrients
Function

Element

Metalloenzyme

Calcium (Ca) Calcium is the most abundant metal in the human body [8,9]. For an average adult, the recommended dietary intake of calcium is 10001200 mg day1 (Table I) depending on age and gender. Interestingly, calcium requirements are biphasic, higher for young adults and elders. Calcium not only promotes several key events, especially haemostasis, but also is a key regulator of epithelialization. Of critical importance to the skin is the role calcium plays in regulating the differentiation of basal keratinocytes to corneocytes [10,11]. Figure 1 schematically shows the relationship of Ca++ with skin barrier homoeostasis. In addition to its role in differentiation, Ca++ appears to be a key modulator of directional locomotion of human keratinocytes and likely promotes wound healing in vivo [12]. Moreover, there is also evidence to suggest that the proper combination of calcium and magnesium (Mg) enhances barrier repair (cf. later section on magnesium benets in topical skin care products). Indeed, calcium gluconate solution or gel is frequently used to treat acid-induced burns [13]. Potassium/Sodium (K/Na) Potassium [14] and sodium [15] represent the dominant cellular and extracellular electrolytes and osmolytes, noted for maintenance of the cellular membrane potential. Table I shows the nutritional goals for these two minerals. For men and women >14 years, the recommendation for potassium is 4700 mg day1. However, because of sodiums ubiquity in the American diet, especially in processed foods, the Dietary Guidelines limits the intake of sodium to <2300 mg day1 for this same age group [2]. With regard to the skin, the benets of bathing in salt water, especially solutions prepared from the Dead Sea salts, have nourished the body and soul of humans since antiquity [15]. As will be discussed subsequently, the putative benets of bathing in Dead Sea salts have been attributed to the magnesium in the sea composition, not the sodium or potassium [16]. Magnesium (Mg) Although not as prevalent as the other three macronutrients, Mg is the fourth most abundant mineral in the body [17]. Over 60% of the bodys magnesium is found in the skeleton, and the remainder is distributed in the cells where it is essential for energy metabolism

Cu

Superoxide Dismutase Lysyl Oxidase Collagen Proline Dioxygenase Cytochrome Oxidase Tyrosinase Cytochrome C Reductase Aconitase Glucose 6-Phosphatase Aconitase Hexokinase Catalase Glutathione Peroxidase Matrix Metalloproteases Numerous Protein Kinases Numerous tRNA synthases DNA/RNA Polymerase

Superoxide decomposition (Antioxidant) Collagen, Elastin synthesis Collagen synthesis Energy production Melanin formation Energy production Intermediary metabolism Intermediary metabolism Intermediary metabolism Intermediary metabolism Peroxide decomposition (Antioxidant) Peroxide decomposition (Antioxidant) Matrix remodelling, Wound healing Signal Transduction t RNA synthesis DNA/RNA synthesis

Fe Mg

Mn Se Zn

have been published to support a specic claim or benet for a given mineral in terms of optimal skin health. The objective of this review is to summarize the scientic evidence available on the benets and risks of topical application of mineral salts. Although humans are exposed to various mineral salts each day, this review will focus on the metal ions identied in the Dietary Guidelines for Americans 2010 as essential to health and well-being. Please note, however, we will not review phosphorus because we believe its role in human nutrition is more systemic. Interaction of metal salts with skin In his authoritative 1995 review on the interactions of metal ions with the skin, Lansdown recognizes 15 elements essential to human nutrition and noted seven elements as carcinogens or potential carcinogens [6]. However, the 2010 Dietary Guidelines for Americans (Table I) recognize only nine minerals as nutritionally essential. Additionally, the most recent edition of the Report on Carcinogens [7] now includes cobalt salts, in addition to the seven noted by Lansdown [6]. It should be recognized that a number of variables may contribute to the safety (or toxicity) of any given mineral including (i) inherent toxicity of the mineral, (ii) dosage of exposure, (iii) frequency of exposure and (iv) genetic background of the individual [7]. For the purpose of this review, we will follow the US Dietary Guidelines that recognize nine minerals essential to human nutrition (Table I), but will follow the general classication system used by Lansdown [6] based largely on their preponderance; namely, macronutrients (e.g. requirements ! 100 g d1; calcium, sodium, potassium, magnesium), micronutrients (e.g. requirements on only several mgs/d; iron, zinc, copper) and trace metals (e.g. minimal requirements not established; selenium). As noted earlier, phosphorus will not be covered in this review.

Figure 1 Relationship of epidermal Ca++ with skin barrier properties. Reprinted with permission from Nature Publishing Group Journal Investigative Dermatology [10].

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Interaction of Mineral Salts with the Skin

T. G. Polefka et al.

and cell replication. Mechanistically, Mg is a cofactor in many metalloenzymes [18]. Table II provides several examples of enzymes that depend on Mg for catalytic activity. According to the 2010 American Dietary Guidelines [2], healthy adult men and women require an intake between 310 and 420 mg day1. Unlike some of the other metal elements, there is little evidence for dietary restrictions [18]. Two well-known consumer products are based on magnesium salts, namely talc (magnesium silicate) and Epsom salts (MgSO4). Talc is most frequently found in baby powders, where it functions as an astringent to prevent diaper rash. Many adults also use baby powders to reduce wetness in the perineal and axilla areas and as a lubricant to enhance skin feel on other areas of the body. Talc can also be found in colour cosmetics and other topical products (i. e. anti-fungal, etc.) where it functions as an anti-caking agent. In recent years, the use of talc has decreased in response to the perceived risk that dusting with talc-containing products causes health problems. This risk was largely attributable to the contamination of the mineral with asbestos. However, a recent critical review of the literature by Muscat and Huncharek found little evidence that talc was carcinogenic, mechanistically or clinically [19]. The other magnesium-containing consumer product is Epsom salts (MgSO4). Epsom salt can be used internally; as a laxative, and topically in bath water to enhance skin softening and exfoliation, relieve muscle tension and to promote relaxation. Although little evidenced-based research is available in the medical literature to support these claims, mechanistically, Mg++ and Ca++ play a key role in regulating keratinocyte proliferation and differentiation and have been shown to activate keratinocyte migration, down-regulating E-cadherin and up-regulating a1b2-integrin function [20]. Denda et al.[21] studied the effects of topical application of magnesium and calcium salts on skin barrier repair in hairless mice. All of the Mg salts, except Mg bis (dihydrogen phosphate), accelerated barrier repair in this animal model. Moreover, optimum barrier repair required a Ca++ to Mg++ ratio less than unity, suggesting a complex and often antagonistic relationship between Mg++ and Ca++ in cornication. In a small human clinical study (n = 12), Schempp et al.[22] showed that topical treatment with 5% MgCl2 prior to UVB irradiation not only signicantly reduced the number of Langerhan cells in the epidermis compared with NaCl, but also reduced antigen-presenting activity (mixed lymphocyte reaction) in the skin in the MgCl2-treated subjects. In contrast to Epsom salt, efforts to understand the medicinal benets of bathing in saline or Dead Sea salts (balneotherapy) are substantial [23]. Interest in balneotherapy is driven by the perceived benets of bathing in the saltiest sea in the world (320 g L1 vs. an average 40 g L1) that also has the highest concentration of Mg (49 g L1) [24]. Moreover, this therapy is consistent with several modern social movements, including complementary medicine and the rediscovery of Spas for relaxation, health and well-being [25]. Despite substantial clinical research, the evidence supporting the medicinal benets of balneotherapy for the treatment of psoriasis must be considered inconclusive. Several human clinical studies [26,27] present ndings that support the efcacy of balneophototherapy for psoriasis. However, these studies have been criticized for study design deciencies. In contrast to these studies, Halevy et al. [28] were unable to discern any meaningful improvements in psoriasis vulgaris when daily soaking in Dead Sea salts was compared with tap water. Several studies also looked at the benet of pre-soaking psoriasis patients in saline or

Dead Sea salts prior to phototherapy. Interestingly, when relevant controls were included in the phototherapy study, the investigators failed to observe meaningful clinical differences [29 31]. To explain these results, Gambichler et al. [32] propose that soaking the skin prior to phototherapy increases UV gain, regardless of salt concentration. In terms of atopic dermatitis, fewer studies are reported in the literature. Proksch and co-workers reported signicant improvement in skin barrier properties (i.e. decreased TEWL, increased hydration, reduced skin roughness and erythema) when patients soaked in 5% Dead Sea salts daily compared with tap water [33]. In a more recent study, Portugal-Cohen et al. [34] explored in a randomized, double-blind vehicle-controlled study involving 86 children with atopic dermatitis, the therapeutic benet of twicedaily application of emollient with and without Dead Sea salts. Efcacy was based on SCORing Atopic Dermatitis (SCORAD), TEWL and several different subjective assessments. Despite observable and measurable improvements in objective and subjective parameters, none of the Dead Sea salt emollient formulations were statistically better than the vehicle control [34]. Taken together, we must conclude that based on the relevant published clinical literature, the evidence supporting the efcacy of balneotherapy for the treatment of psoriasis and atopic dermatitis is inconclusive. However, this treatment does appear to offer patients a palliative benet in term of alleviating some symptoms (e.g. pruritus, akiness, etc.) of these conditions. For a more thorough discussion on this complementary therapy, the reader is referred to the following reviews [35,36]. Micronutrients Zinc (Zn) The daily requirement for zinc in men and women is 11 and 8 mg day1, respectively (Table I). Interestingly, approximately 11% of the bodys Zn is associated with the epidermis [37]. From a biochemical point of view, Zn represents a dominant cofactor for many enzymes (Table II) [38,39], including those involved in wound healing [40], and most recently has been hypothesized to function as a non-traditional antioxidant [41]. Although Zn appears to be safe and effective in many topical applications, it is not totally innocuous. For example, in woundhealing studies, topical application of ZnCl2 and ZnSO4 were not only irritating, but also delayed barrier repair [40]. However, despite several exceptions, Zn compounds appear to offer the greatest therapeutic potential of all the metal minerals as evidence by the number of FDA-approved usages [35]. Table III lists the four product categories and zinc compounds approved for over-thecounter (OTC) uses [4247]. In the United States, the FDA recognizes three Zn compounds: zinc acetate (0.12%), zinc carbonate (0.22%) and zinc oxide (1 25%), as safe and effective for use as topical skin protectants [45]. However, despite the opportunity to use the carbonate and acetate salts of zinc, almost all skin protectants are based on ZnO, probably because it is cost-effective, easily formulated and stable under aerobic conditions. According to Lansdown et al. [40], the medicinal properties of zinc, in the form of calamine, were rst documented more than 2000 years ago in the Ebers Papyrus. Calamine is a mixture of ZnO with approximately 0.5% Fe2O3 [45]. As noted earlier, zinc is an essential micronutrient. Requirements are satised by a

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Interaction of Mineral Salts with the Skin

Table III Use of Zinc salts and compounds approved by the FDA for overthe-counter human use

T. G. Polefka et al.

Product

Compound

Indication

Conc. (%)

Ref

Anti-fungal

Zn undecylenate

Dandruff and Seborrhoea Dermatitis

Zn pyrithione

Skin Protection

Zn acetate

Zn carbonate

Zn oxide

Sun Protection

Zn Oxide

For the treatment of athletes foot, jock itch and ringworm For the control or relief of dandruff and/or seborrhoeic dermatitis. To control akes and scalp itch Skin protectant Promotes healing of minor skin irritation and sunburn Protects chafed skin associated with diaper rash and helps protect from wetness Dries the oozing and weeping of poison ivy, oak, and sumac, insect bites. Helps to prevent sunburn

1025

36

0.12.0

37

0.12.0

38

0.22.0

40

1.025

58

Up to 25

39

well-balanced diet, leading to an average daily intake of 10 15 mg day1, consistent with a recommended daily allowance of 8 mg day1 and 11 mg day1 for women and men, respectively (Table I). Clinical deciencies of Zn were rst reported in 1961 [37]. However, from a public health perspective, this deciency appears to be limited to developing countries. Approximately 50% of the available Zn is localized to the cytoplasm, 3040% in the nucleus, and the remainder is associated with the plasma membrane [37]. Within the cytoplasm, approximately 20% is associated with the zinc-binding protein, metallothioneins (MTs); MTs are low molecular weight proteins (67 Kd) with a high (approximately 30%) cysteine content that appear to function as Zn and copper (Cu) storage molecules [48]. Although the MTs are expressed constitutively in skin cells, expression is signicantly up-regulated in cells exhibiting high mitotic activity such as those found in wound margins. Importantly, Zn is a cofactor in numerous biochemical reactions. Indeed, the prevalence of genes encoding zinc proteins is estimated at over 3% of the 32 000 identied genes [49]. According to Schwartz et al. [39], over 300 Zndependent enzymes have been identied and characterized. Table II lists several examples. Since the early phases of wound healing requires the action of metalloproteases, it is believed that MTs function as a source of Zn++ and Cu++ necessary for metalloenzyme synthesis [48]. In human skin, the Zn concentration in the epidermis (50 70 lg g1 dry weight) is higher than in the dermis (1015 lg g1 dry weight) [39]. Within the epidermis, the Zn++ concentration gradient is inverted to Ca++, with higher levels localized to

metabolically active basal cell layer and much less at the stratum corneum [48]. This is not surprising because low Ca++ and high Zn++ stimulate keratinocyte proliferation [10,11]. Studies in experimental wound models suggest that supplemental Zn enhances wound healing [40,49]. In the rat model, Zn levels in the wound margin increase 1520% within 24 h and increase to 30% by the time re-epithelialization begins. Although zincs anti-microbial activity may accelerate healing, this hypothesis is not universally accepted. As noted above, proliferating cells require Zn++ and metallothioneins for the production of the Zn-requiring metalloproteinases (MMPs), RNA and DNA polymerases and many other Zn-containing enzymes. Some wound healing research suggests that the manner in which Zn is presented to the tissue may be important. Agren et al. [50] found ZnO to be superior to ZnSO4 in terms of mitigating inammation and enhancing re-epithelialization of partial thickness porcine skin. The authors attributed the efcacy of ZnO to its lower water solubility, and ability to provide sustained release of Zn++ to the wound site at non-toxic levels. It is worth noting that the aqueous solubility of ZnO is much lower than almost any other Zn salt with the exception of ZnCO3 (ZnO Ksp = 3.86 9 1010 vs. ZnCO3 Ksp = 1.4 9 1011) [51]. In a double-blind, placebo-controlled human clinical study, topically applied ZnO signicantly promoted healing of leg ulcers [52]. Others investigators recognize ZnO as a topical debriding agent for pressure ulcer [53, 54] and as an occlusive dressing for diabetic foot ulcers [55]. In smaller clinical studies, investigators claim ZnO enhances healing of burn wounds [56], suction-blister wounds [57], supercial (i.e. 1 mm deep) small incisions [58] and the epithelialization donor graft sites [59]. However, as noted by Lansdown [40] larger-scale trials are urgently required to verify the benets of topical zinc oxide in acute human wounds. The percutaneous penetration of ZnO has been studied [60, 61]. Although the FDA accepts the concept that penetration of ZnO through intact skin is extremely low [45], abrogation of the stratum corneum barrier enhances the penetration of Zn because of increased hydration. Most recently, Newman et al. [62] reviewed the safety of nanosized titanium dioxide and zinc oxide particles, as it relates to sunscreen products, and concluded Although we found no evidence of signicant penetration of titanium dioxide and zinc oxide nanosized particles beyond the stratum corneum, further studies must be carried out to simulate real-world conditions particularly in sunburned skin and under ultraviolet exposure. ZnO has been used in topical products for quite some time. As noted previously, one of the rst FDA-approved usages of ZnO is as a skin protectant [45]. Accordingly, products containing between 1% and 25% ZnO can claim on their label to be a poison ivy, oak, sumac protectant and to dry the oozing and weeping of poison ivy, oak, sumac. Additionally, ZnO is approved as a broad-spectrum sunscreen agent [66]. Unlike the organic sunscreens, the mineral sunscreens such as ZnO and TiO2 not only reduce skin penetration by UVB radiation, but also provide protection against UVA radiation. As noted in Table III, ZnO can be used in topical sunscreen formulations up to 25% [63]. Interestingly, consumer interest in sunscreen products based on ZnO and TiO2 has grown in recent years because of heightened consumer awareness of the damage to skin caused by increasing exposure to solar radiation and has led to the marketing of several commercial sunscreen products based solely on these mineral sunscreens. Clearly, ZnO has a long history of safe and effective use and is recognized for skin rashes such as diaper rash, prickly heat and skin conditions such as eczema, impetigo, ringworm, ulcers, pruritus and psoriasis [45].

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Interaction of Mineral Salts with the Skin

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Iron (Fe) Iron is one of the most abundant trace metals in the body and functions largely in oxygen transport and oxidationreduction reactions, especially in respiration [64, 65]. As can be seen in Table II, Fe plays a role in various oxygenases, including the skin-relevant procollagen-proline dioxygenase. One estimate suggests that 70% of the bodys Fe is associated with haemoglobin [6]. Levels in normal epidermis and psoriatic epidermis vary over a broad range [66, 67]. Much like calcium, the bodys requirement for iron varies with gender and age (Table I). For adult males, the daily iron requirement is 8 mg. Actively menstruating women require 1518 mg day1, whereas post-menopausal women require signicantly less, only 8 mg day1 [1]. Toxicity of Fe appears route specic. Higdon and Drake [65] note that accidental overdose of iron-containing products is the single largest cause of poisoning fatalities in children <6 year of age. In contrast to this, colour cosmetics containing iron oxide have been applied to the facial skin to beautify and/or camouage minor imperfections since antiquity [68, 69]. Although there are few reports on the topical toxicity of iron or iron compounds, there is signicant evidence from experimental animal models implicating the release of Fe from haemoglobin by UVR in the induction of cutaneous oxidative stress [70, 71]. Interestingly, iron analysis of epidermis derived from sun-exposed and unexposed skin revealed that the sun-exposed skin exhibited signicantly higher levels of free iron than unexposed skin (53.0 vs. 17.8 ppm), respectively [69]. In subsequent work, Bissett and McBride showed in guinea pig and mouse models that topical application of an iron chelator (2-furildioxime) signicantly delayed UVB-induced tumour onset [70]. To the best of our knowledge, we are unaware of any specic therapeutic usage of topical iron delivery. Copper (Cu) Copper is an important trace mineral found throughout the body where it serves as a cofactor for several enzymes, including lysyl oxidase, the enzyme involved in cross-linking collagen, and tyrosinase, the enzyme involved in skin pigmentation [72]. Indeed, at least one study reports the benets of dietary supplementation on this enzyme in skin [73]. The recommended daily allowance for Cu for healthy adult men and women over the age of 19 is 900 lg day1 (Table I). Although deciencies of Cu appear rare, conditions that give rise to intestinal malabsorption (i.e. coeliac disease, bowel resection, etc.) or defective transport of Cu is frequently diagnosed because of the visible changes in skin pigmentation and/ or hair growth (i.e. Menkes kinky hair syndrome) [74]. Conversely, excessive exposure to Cu compounds is easily diagnosed by the characteristic green hue it produces in hair [74]. In 1973, Pickard and Thaler [75] isolated a tripeptidecopper complex from serum that enhanced collagen formation in cultured cells. Several commercial cosmetic products are based on a proprietary copperpeptide complex (Cu-GHK, Cu-AHK). According to a website managed by Loren Pickart, PhD, copperpeptides have been proven to calm irritated skin, improve skin elasticity and rmness, repair photodamaged skin, reduce ne lines and wrinkles, accelerate wound healing and a host of other benets [76]. Recently, Mazurowska and Mojski [77] reported on the ability of GHK-Cu and GSH-Cu to penetrate liquid crystalline liposomal membranes. Interestingly, using the same tripeptides, but with manganese as the metal ion, Hussain and Goldberg [78] report

that topical application of a serum formulation twice daily for 12 weeks, signicantly improved the visual signs of photoaged skin, especially hyperpigmentation. Other applications of Cu compounds are related to their wellknown antibacterial, anti-fungal and antiviral activities [79]. Borkow et al. recently reported a more provocative use of Cu [80]. In a 4-week double-blinded, randomized study involving 57 volunteers, this team observed that subjects sleeping on pillowcases containing 0.4% copper oxide exhibited signicantly reduced facial wrinkles, crows feet/lines and a global improvement in appearance. The authors attribute the improvement in skin appearance to coppers ability to penetrate the skin and stimulate the formation of extracellular matrix proteins. Topical use of Cu appears limited because of its potential to induce oxidationreduction reactions and general toxicity. However, one cosmetic company recently created a bimineral complex composed of copper and zinc that is claimed to generate a galvanic signal capable of reducing inammation [81]. In a double-blind, placebo-controlled study, Chantalat et al. [82, 83] compared the performance of a placebo formula (gel + activating moisturizer) to an active system (bimineral complex gel + activating moisturizer) to improve the appearance of individuals with photoaged skin. Although both treatments signicantly (P < 0.05) improved visual signs of ageing from baseline, the bimineral complex formulation provided signicantly greater improvement than the vehicle control (P < 0.05) for attributes such as skin tone and colour, skin texture, ne lines, crows feet wrinkles, wrinkles and dark circles around the eyes. Despites its attractiveness from a marketing perspective, we believe more rigorous clinical studies are necessary to establish the value of galvanic treatments. Selenium (Se) Selenium is commonly found in the soil, particularly, in Western United States, where it accumulates in plants as selenomethionine and selenocysteine [84]. In humans, these two amino acids are key components of antioxidant enzymes such as glutathione peroxidase and thioredoxin reductase [85]. Although deciencies in humans are rare, animal studies have implied a possible link between selenium deciency and cancer [86]. Oral administration of Se to mice has been shown to mitigate UVR-induced inammation, pigmentation, hyperkeratosis and carcinogenesis [87, 88]. In a small human clinical study with 8 subjects, topical application of lotions containing selenomethionine (0.0020.05%) the most effective way of delivering selenium into the skin for 2 weeks was shown to mitigate the acute effects of UV exposure [88]. In addition to inhibiting photodamage, research in animal models suggests that selenomethionine may also reverse photoageing [89]. However, in a recently published Cochrane Review, Dennert et al. [90] systematically reviewed 55 studies involving more than one million participants and was not able to establish reliable conclusions regarding a causal relationship between low selenium exposure and increased the risk of cancer and the benets of selenium supplementation in reducing the incidence of cancer in humans. Mechanistically, Se is believed to protect the skin and other organs through its involvement in antioxidant enzymes, especially glutathione peroxidase and thioredoxin reductase [91]. Rafferty et al.[92] have shown that human broblasts, keratinocytes and melanocytes express between 10 and 15 selenoproteins. Supplementation of culture media with either selenium or

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selenomethionine signicantly reduced UV-induced death of keratinocytes and melanocytes [92]. In a more recent study, Sengupta et al.[93] used a mouse model where the Sec tRNA (the unique tRNA that codes for selenocysteine) was knocked out, limiting the animals ability to make selenoproteins. The results of this deciency included epidermal hyperplasia, aberrant hair follicle development progressing to alopecia and ultimately to premature death [93]. Taken together, these results support the crucial role of selenium and selenoproteins in the well-being of skin. In the United States, topical application of selenium sulphide is approved in rinse-off products for the treatment of dandruff and seborrhoeic dermatitis [44]. Trace elements Interestingly, the Dietary Guidelines for Americans, 2010 [2] do not establish recommendations for dietary intakes for any of the trace metals (i.e. manganese, chromium, molybdenum, etc.). However, one should not interpret the absence of an RDI to mean that these trace elements are unimportant. Non-nutrient metal elements In addition to the metal elements recognized for their nutritional value, several others (i.e. aluminium, zirconium, silver, gold, mercury and tin) are frequently found in the human body. However, except for aluminium and zirconium salts that represent the active ingredients in topic antiperspirant products [94,95], contact with

most of these metal ions is attributed to environmental exposure [6]. Conclusion Human skin is the largest organ of the integumentary system and is made up of multiple layers of ectodermal tissue. It is a dynamic organ containing complex biological processes. Our skin interfaces with the environment, where its serves as a barrier that protects the body against pathogens, foreign bodies, water loss, solar radiation and has a key role in temperature regulation, sensation and vitamin D production. The skin is dependent on the systemic circulatory system to supply it with nutrients and thus reects systemic nutritional deciencies. For individuals with adequate nutritional status, the question remains: what value does topical supplementation provides to overall skin health? The cosmetic industry has demonstrated the benets for topical supplementation of the skin with vitamins such as vitamin A, vitamin C, vitamin E and nicotinamide as well as numerous other natural antioxidants. However, evidences supporting the benets of minerals have been limited to several such as ZnO, TiO2 and Se (anti-fungal agent). Although there is much work published on Mg-rich Dead Sea salts, Cu and Se, additional rigorous, hypothesis-driven clinical studies are necessary. Acknowledgement Merck Consumer Care funded this work.
16. Matz, H., Orion, E. and Wolf, R. Balneotherapy in dermatology. Dermatol. Ther. 16, 132 140 (2003). 17. Fawcett, W., Haxby, E. and Male, D. Magnesium: physiology and pharmacology. Br. J. Anaesth. 83, 302320 (1999). 18. Higdon, J. and Drake, V.J. Magnesium. An Evidence-Based Approach to Vitamins and Minerals, 2nd edn. pp. 169178. Georg Thieme Verlag, Stuttgart, Germany (2012). 19. Muscat, J.E. and Huncharek, M.S. Perineal talc use and ovarian cancer: a critical review. Eur. J. Cancer Prev. 17, 139146 (2008). ` ve20. Boisseau, A.-M., Donatien, P., Surle Bazeille, J.-E., et al. Production of epidermal sheets in a serum free culture system: a further appraisal of the role of extracellular calcium. J. Dermatol. Sci. 3, 111120 (1992). 21. Denda, M., Katagiri, C., Hirao, T., Maruyama, N. and Takahashi, M. Some magnesium salts and a mixture of magnesium and calcium salts accelerate skin barrier recovery. Arch. Dermatol. Res. 291, 560 563 (1999). 22. Schempp, C.M., Dittmar, H.C., Hummler, D., et al. Magnesium ions inhibit the antigenpresenting function of human epidermal Langerhans cells in vivo and in vitro: involvement of ATPase, HLA-DR, B7

References
1. RNCOS. Global Cosmeceuticals Market Analysis. Available at: http://www.rncos. com/Report/IM366.htm (accessed 13 April 2012). 2. Dietary Guidelines for Americans, 2010, 7th edn. U.S. Government Printing Ofce, Washington, DC (2010). 3. Voet, D. and Voet, J.G. Biochemistry, 4th edn. p. 1520. John Wiley & Sons, New York (2010). 4. Zussman, J., Ahdout, J. and Kim, J. Vitamins and photoaging: do scientic data support their use? J. Am. Acad. Dermatol. 63, 507525 (2010). 5. Thornfeldt, C.R. Cosmeceuticals: separating fact from voodoo science. SKINmed 4, 214 220 (2005). 6. Lansdown, A.B.G. Physiological and toxicological changes in the skin resulting from the action and interaction of metal ions. CRC Crit Rev Toxicol. 25, 397462 (1995). 7. 11th Report on Carcinogens (RoC). National Toxicology Program, Washington, DC (2005). 8. Lansdown, A.B.G. Calcium: a potential central regulator in wound healing in the skin. Wound Repair Regen. 10, 271285 (2002). 9. Higdon, J. and Drake, V.J. Calcium. An Evidence-Based Approach to Vitamins and 10.

11.

12.

13.

14.

15.

Minerals, 2nd edn. pp. 115127. Georg Thieme Verlag, Stuttgart, Germany (2012). Elias, P.M., Ahn, S.K., Brown, B.E., Crumrine, D. and Feingold, K.R. Origin of the epidermal calcium gradient: regulation by barrier status and role of active vs. passive mechanisms. J Invest Dermatol. 119, 12691274 (2002). Elias, P.M., Ahn, S.K., Denda, M., et al. Modulations in epidermal calcium regulate the expression of differentiation-specic markers. J Invest Dermatol. 119, 1128 1136 (2002). Fang, K.S., Farboud, B., Nuccitelli, R. and Isseroff, R.R. Migration of human keratinocytes in electric elds requires growth factors and extracellular calcium. J Invest Dermatol. 111, 751756 (1998). Trevin o, M.A., Herrmann, G.H. and Sprout, W.L. Treatment of severe hydrouoric acid exposures. J. Occup. Environ. Med. 25, 861 863 (1983). Higdon, J. and Drake, V.J. Potassium. An Evidence-Based Approach to Vitamins and Mineral, 2nd edn. pp. 196213. Georg Thieme Verlag, Stuttgart, Germany (2012). Higdon, J. and Drake, V.J. Sodium (Chloride). An Evidence-Based Approach to Vitamins and Minerals, 2nd edn. pp. 214223. Georg Thieme Verlag, Stuttgart, Germany (2012).

2012 Copyright 2009-2012 Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Whitehouse Station, N.J., U.S.A. All rights reserved. International Journal of Cosmetic Science, 34, 416423 421

Interaction of Mineral Salts with the Skin

T. G. Polefka et al.

23.

24.

25.

26.

27.

28.

29.

30.

31.

32.

33.

molecules, and cytokines. J Invest Dermatol. 115, 680686 (2000). Nasermoaddeli, A. and Kagamimori, S. Balneotherapy in medicine: a review. Environ. Health Prev. Med. 10, 171179 (2005). Charlier, R. and Chaineux, M.-P. The healing sea: a sustainable Coastal Ocean resource: thalassotherapy. J. Coastal Res. 25, 838856 (2009). Riyaz, N. and Arakkal, F. Spa therapy in dermatology. Indian J Dermatol, Venereol Leprol. 77, 128134 (2011). Harari, M., Czarnowicki, T., Fluss, R., Ruzicka, T. and Ingber, A. Patients with early-onset psoriasis achieve better results following Dead Sea climatotherapy. J. Eur. Acad. Dermatol. Venereol. 65, 525530 (2011). Klein, A., Schiffner, R., Schiffner-Rohe, J., et al. A randomized clinical trial in psoriasis: synchronous balneophototherapy with bathing in Dead Sea salt solution plus narrowband UVB vs. narrowband UVB alone (TOMESA-study group). J. Eur. Acad. Dermatol. Venereol. 25, 570578 (2011). Halevy, S., Giryes, H., Friger, M. and Sukenik, S. Dead sea bath salt for the treatment of psoriasis vulgaris: a double-blind controlled study. J. Eur. Acad. Dermatol. Venereol. 9, 237242 (1997). Gambichler, T., Rapp, S., Senger, E., Altmeyer, P. and Hoffmann, K. Balneophototherapy of psoriasis: highly concentrated salt water versus tap water a randomized, one-blind, right/left comparative study. Photodermatol. Photoimmunol. Photomed. 17, 2225 (2001). Brockow, T., Schiener, R., Franke, A., Resch, K. and Peter, R. A pragmatic randomized controlled trial on the effectiveness of low concentrated saline spa water baths followed by ultraviolet B (UVB) compared to UVB only in moderate to severe psoriasis. J. Eur. Acad. Dermatol. Venereol. 21, 1027 1037 (2007). Brockow, T., Schiener, R., Franke, A., Resch, K. and Peter, R. A pragmatic randomized controlled trial on the effectiveness of highly concentrated saline spa water baths followed by UVB compared to UVB only in moderate to severe psoriasis. J. Altern. Complement. Med. 13, 725732 (2007). Gambichler, T., Demetriou, C., Terras, S., Bechara, F.G. and Skrygan, M. The impact of salt water soaks on biophysical and molecular parameters in psoriatic epidermis equivalents. Dermatology 223, 230238 (2011). Proksch, E., Nissen, H.-P., Bremgartner, M. and Urquhart, C. Bathing in a magnesiumrich Dead Sea salt solution improves skin barrier function, enhances skin hydration,

34.

35.

36.

37.

38.

39.

40.

41.

42.

43.

44.

45.

46.

47.

and reduces inammation in atopic dry skin. Intl J Dermatol. 44, 151157 (2005). Portugal-Cohen, M., Oron, M., Merrik, E., et al. A Dead Sea water-enriched body cream improves skin severity scores in children with atopic dermatitis. J Cosmet, Dermatol Sci Appl. 1, 7178 (2011). Falagas, M.E., Zarkadoulia, E. and Rafailidis, P.I. The therapeutic effect of balneotherapy: evaluation of the evidence from randomized controlled trials. Int. J. Clin. Pract. 63, 1068 1084 (2009). Smith, N., Weymann, A., Tausk, F.A. and Gelfand, J.M. Complementary and alternative medicine for psoriasis: a qualitative review of the clinical trial literature. J. Am. Acad. Dermatol. 61, 841856 (2009). Higdon, J. and Drake, V.J. Zinc. An EvidenceBased Approach to Vitamins and Minerals, 2nd edn. pp. 224233. Georg Thieme Verlag, Stuttgart, Germany (2012). Tapiero, H. and Tew, K.D. Trace elements in human physiology and pathology: zinc and metallothioneins. Biomed. Pharmacother. 57, 399411 (2003). Schwartz, J.R., Marsh, R.G. and Draelos, Z. D. Zinc and skin health: overview of physiology and pharmacology. Dermatol. Surg. 31, 837847 (2005). Lansdown, A.B.G., Mirastschijski, U., Stubbs, gren, M.S. Zinc in N., Scanlon, E. and A wound healing: theoretical, experimental, and clinical aspects. Wound Repair Regen. 15, 216 (2007). Powell, S.R. Zinc and Health: current Status and Future Directions. J. Nutr. 130, 1447S 1454S (2000). Status of OTC Rulemakings. FDA; 2011; Available at: http://www.fda.gov/Drugs/ DevelopmentApprovalProcess/Development Resources/Over-the-CounterOTCDrugs/Status ofOTCRulemakings/default.htm, accessed 17 February 2011 FDA, Topical antifungal drug products for over-the-counter human use; nal monograph. Fed Regist. 58: 4989049899 (1989). FDA, Dandruff, seborrheic dermatitis, and psoriasis drug products for over-the-counter human use, nal monograph. Fed Regist. 56, 6355463569 (1991). FDA, Skin protectant drug products for overthe-counter human use; nal monograph. Fed Regist. 68, 3336233381 (2003). FDA, Sunscreen drug products for over-thecounter human use; nal monograph. Fed Regist. 64, 2766627693 (1999). FDA, Skin protectant drug products for over-the-counter human use; advanced proposed rule. Fed Regist. 43, 3462862848 (1978).

48. Lansdown, A.B.G. Metallothioneins: potential therapeutic aids for wound healing in the skin. Wound Repair Regen. 10, 130132 (2002). 49. Berg, J.M. and Shi, Y. The galvanization of biology: a growing appreciation for the roles of zinc. Science 271, 10811085 (1996). gren, M.S., Chvapil, M. and Franze n, L. 50. A Enhancement of re-epithelialization with topical zinc oxide in porcine partial-thickness wounds. J. Surg. Res. 50, 101105 (1991). 51. Solubility products of selected compounds. Salt Lake Metals; Available at: http://www.silver nitrate.info/SolubilityProducts.htm (Accessed 16 May 2011). 52. Agren, M.S. Studies on zinc in wound healing. Acta. Derm. Venereol. Suppl. (Stockh). 154, 136 (1990). gren, M.S. Topical 53. Strbero mg, H.E. and A zinc oxide treatment improves arterial and venous leg ulcers. Br. J. Dermatol. 111, 461 468 (1984). gren, M.S. and Stro 54. A mberg, H.E. Topical treatment of pressure ulcers: a randomized comparative trial of Varidase and zinc oxide. Scand. J. Plast. Reconstr. Surg. Hand Surg. 19, 97100 (1985). 55. Apelqvist, J., Larsson, J. and Lstenstrom, A. Topical treatment of necrotic foot ulcers in diabetic patients: a comparative trial of DuoDerm and MeZinc. Br. J. Dermatol. 123, 787 792 (1990). 56. Gang, R. Adhesive zinc tape in burns: results of a clinical trial. Burns. 7, 322325 (1981). 57. Tronnier, H. Experimental study on epithelization. Z Hautkr. 50, 925929 (1975). 58. Greenway, S., Filler, L. and Greenway, F. Topical insulin in wound healing: a randomized, double-blind, placebo-controlled trial. J Wound Care. 8, 526528 (1999). 59. Weingart, D. and Stoll, P. The epithelialization of split skin graft donor sites a test model for the efcacy of topical wound therapeutic agents. Eur J of Plast Surg. 16, 22 25 (1993). gren, M.S. Percutaneous absorption of zinc 60. A from zinc oxide applied topically to intact skin in man. Dermatol. 180, 3639 (1990). 61. Lansdown, A.B.G. and Taylor, A. Zinc and titanium oxides: promising UV-absorbers but what inuence do they have on the intact skin? Int. J. Cosmet. Sci. 19, 167172 (1997). 62. Newman, M.D., Stotland, M. and Ellis, J.I. The safety of nanosized particles in titanium dioxide-and zinc oxide-based sunscreens. J. Am. Acad. Dermatol. 61, 685692 (2009). 63. Sunscreen drug products for over-the-counter human use; amendment to the tentative

2012 Copyright 2009-2012 Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Whitehouse Station, N.J., U.S.A. All rights reserved. 422 International Journal of Cosmetic Science, 34, 416423

Interaction of Mineral Salts with the Skin

nal monograph. Fed Regist. 63, 56584 56589 (1998). Lansdown, A.B.G. Iron: a cosmetic constituent but an essential nutrient for healthy skin. Int. J. Cosmet. Sci. 23, 129137 (2001). Higdon, J. and Drake, V.J. Iron. An Evidenced-Based Approach to Vitamins and Minerals, 2nd edn. pp. 157168. Georg Thieme Verlag, Stuttgart, Germany (2012). Molin, L. and Wester, P. Iron content in normal and psoriatic epidermis. Acta Derm. Venereol. 53, 473476 (1973). Kurz, K., Steigleder, G.K., Bischof, W. and Gonsior, B. PIXE analysis in different stages of psoriatic skin. J Invest Dermatol. 88, 223 226 (1987). Draelos, D. Colored facial cosmetics. Dermatol. Clin. 18, 621631 (2000). Antoniou, C. and Stefanaki, C. Cosmetic camouage. J Cosmet Dermatol. 5, 297301 (2006). Bissett, D.L. and McBride, J.F. Iron content of human epidermis from sun-exposed and non-exposed body sites. J. Soc. Cosmet. Chem. 43, 215217 (1992). Bissett, D.L. and McBride, J.F. Synergistic topical photoprotection by a combination of the iron chelator 2-furildioxime and sunscreen. J. Am. Acad. Dermatol. 35, 546549 (1996). Higdon, J. and Drake, V.J. Copper. An Evidence-Based Approach to Vitamins and Minerals, 2nd edn. pp. 135141. Georg Thieme Verlag, Stuttgart, Germany (2012). Werman, M.J., Bhathena, S.J. and Turnlund, J.R. Dietary copper intake inuences skin lysyl oxidase in young men. J. Nutr. Biochem. 8, 201204 (1997). de Roma, D.L., Olivares, M., Uauy, R. and Araya, M. Risks and benets of copper in light of new insights of copper homeostasis. J. Trace Elem. Med Biol. 25, 313 (2011). Pickart, L., Thayer, L. and Thaler, M. A synthetic tripeptide which increases survival of normal liver cells, and stimulates growth in hepatoma cells. Biochem. Biophys. Res. Commun. 54, 562566 (1973).

T. G. Polefka et al.
(Dayan, N., ed.) pp. 149176. William Andrew Inc, Norwich, NY (2008). Ip, C. Lessons from basic research in selenium and cancer prevention. J. Nutr. 128, 18451854 (1998). Burke, K.E. and Calvo, L.C. Topical compositions containing selenoamino acids for the prevention of ultraviolet radiation-induced skin damage. United States patent 4,865,480. September 12, (1989). Burke, K.E. Oral and topical, L. selenomethionine protection from ultraviolet-induced sunburn, tanning and skin cancer. J Orthomol Med. 7, 8394 (1992). Burke, K., Burford, R., Combs, G., French, I. and Skefngton, D. The effect of topical L-selenomethionine on minimal erythema dose of ultraviolet irradiation in humans. Photodermatol. Photoimmunol. Photomed. 9, 5257 (1992). Dennert, G., Zwahlen, M., Brinkman, M., Vinceti, M., Zeegers, M. and Horneber, M.. Selenium for preventing cancer (Review). The Cochrane Library. (5): 1126 (2011). Pence, B.C., Delver, E. and Dunn, D.M. Effects of dietary selenium on UVB-induced skin carcinogenesis and epidermal antioxidant status. J Invest Dermatol. 102, 759 761 (1994). Rafferty, T.S., McKenzie, R.C., Hunter, J., et al. Differential expression of selenoproteins by human skin cells and protection by selenium from UVB-radiation-induced cell death. Biochem J. 332, 231236 (1998). Sengupta, A., Lichti, U.F., Carlson, B.A., et al. Selenoproteins are essential for proper keratinocyte function and skin development. PLoS 5, e12249 (2010). FDA. Antiperpsirant drug products for overthe-counter human use, nal monograph. Fed Regist. 68: 3427334293 (2003). Abrutyn, E. Antiperspirant and deodorants. In: Cosmetic Dermatology: Products & Procedures (Draelos, Z., ed.), pp. 150155. WileyBlackwell, Oxford, UK and Hoboken, NJ (2010).

64.

65.

66.

67.

68. 69.

70.

71.

72.

73.

74.

75.

76. Pickart, L. Published studies on tissue and skin remodeling copper-peptides. Available at: http://skinbiology.com/ (accessed 29 June 2012). 77. Mazurowska, L. and Mojski, M. Biological activities of selected peptides: skin penetration ability of copper complexes with peptides. J. Soc. Cosmet. Chem. 59, 5969 (2008). 78. Hussain, M. and Goldberg, D.J. Topical manganese peptide in the treatment of photodamaged skin. J Cosmet Laser Ther. 9, 232236 (2007). 79. Borkow, G. and Gabbay, J. Copper, An ancient remedy returning to ght microbial, fungal and viral infections. Cur Chem Biol. 3, 272278 (2009). 80. Borkow, G., Gabbay, J., Lyakhovitsky, A. and Huszar, M. Improvement of facial skin characteristics using copper oxide containing pillowcases: a double blind, placebo controlled, parallel, randomized study. Int. J. Cosmet Sci. 31, 437443 (2009). 81. Kaur, S., Lyte, P., Garay, M., et al. Galvanic zinccopper microparticles produce electrical stimulation that reduces the inammatory and immune responses in skin. Arch. Dermatol. Res. 303, 551562 (2011). 82. Chantalat, J., Bruning, E., Sun, Y. and Liu, J.-C. Biomimetic signaling technology generated by a bi-mineral complex demonstrates efcacy in reducing clinical photo-aging in a 12-week placebo-controlled study. J. Am. Acad. Dermatol. 62(3 Suppl), AB6 (2010). 83. Chantalat, J., Bruning, E., Liu, J.-C. and Sun, Y. Biomimetic signaling technology generated by a bimineral complex reduces signs of photoaging in the eye area. J. Am. Acad. Dermatol. 62(3 Suppl 1), AB60 (2010). 84. Higdon, J. and Drake, V.J. Selenium. An Evidence-Based Approach to Vitamins and Minerals, 2nd edn. pp. 203213. Georg Thieme Verlag, Stuttgart, Germany (2012). 85. Burke, K.E. Prevention and treatment of aging skin with topical antioxidants. In: Skin Aging Handbook: An Integrated Approach to Biochemistry and Product Development

86.

87.

88.

89.

90.

91.

92.

93.

94.

95.

2012 Copyright 2009-2012 Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Whitehouse Station, N.J., U.S.A. All rights reserved. International Journal of Cosmetic Science, 34, 416423 423