Relevance of vitamins C and E in cutaneous photoprotection

Ultraviolet (UV) radiation-induced oxidative stress may result in acute and chronic photodamage. Based on the endogenous antioxidant system, the administration of antioxidants for scavenging reactive oxygen species might be a promising strategy in the prevention of UV-induced skin reactions. The relevance of the most common antioxidants, vitamins E and C, is reviewed focusing on topical and systemic photoprotective effects in animals and humans. Topically applied vitamin C induced significant photoprotective effects at concentrations of at least 10% in animals and humans, whereas a photoprotective effect has not been demonstrated by oral administration even at high doses in humans. Topical vitamin E reduced erythema, sunburn cells, chronic UV-B-induced skin damage, and photocarcinogenesis in the majority of the published studies, whereas only high doses of oral vitamin E may affect the response to UV-B in humans. Combination of vitamins C and E, partly with other photoprotective compounds, did increase the photoprotective effects dramatically compared to monotherapies. This synergistic interplay of several antioxidants should be taken into consideration in future research on cutaneous photoprotection.     

Topical application of green and white tea extracts provides protection from solar-simulated ultraviolet light in human skin

Background:  Tea polyphenols have been found to exert beneficial effects on the skin via their antioxidant properties.
Aims:  We sought to determine whether topical application of green tea or white tea extracts would prevent simulated solar radiation-induced oxidative damages to DNA and Langerhans cells that may lead to immune suppression and carcinogenesis.
Methods:  Skin samples were analysed from volunteers or skin explants treated with white tea or green tea after UV irradiation. In another group of patients, the in vivo immune protective effects of green and white tea were evaluated using contact hypersensitivity to dinitrochlorobenzene.
Results:  Topical application of green and white tea offered protection against detrimental effects of UV on cutaneous immunity. Such protection is not because of direct UV absorption or sunscreen effects as both products showed a sun protection factor of 1. There was no significant difference in the levels of protection afforded by the two agents. Hence, both green tea and white tea are potential photoprotective agents that may be used in conjunction with established methods of sun protection.     

Low molecular weight antioxidants and their role in skin ageing.

There is increasing evidence that reactive oxygen species play a pivotal role in the process of ageing. The skin, as the outermost barrier of the body, is exposed to various exogenous sources of oxidative stress, in particular UV-irradiation. These are believed to be responsible for the extrinsic type of skin ageing, termed photo-ageing. It therefore seems reasonable to try to increase levels of protective low molecular weight antioxidants through a diet rich in fruits and vegetables or by direct topical application. Indeed, various in vitro and animal studies have proved that low molecular weight antioxidants, especially vitamins C and E, ascorbate and tocopherol, as well as lipoic acid, exert protective effects against oxidative stress. However, controlled long-term studies on the efficacy of low molecular weight antioxidants in the prevention or treatment of skin ageing in humans are still lacking.     

UV photoprotection by combination topical antioxidants vitamin C and vitamin E

BACKGROUND: Virtually all plants and animals protect themselves from the sun using vitamins C and E. OBJECTIVE: The purpose of this study was to see if a combination of topical vitamins C and E is better for UV protection to skin than an equivalent concentration of topical vitamin C or E alone. METHODS: We developed a stable aqueous solution of 15% L-ascorbic acid (vitamin C) and 1% alpha-tocopherol (vitamin E). We applied antioxidant or vehicle solutions to pig skin daily for 4 days. We irradiated (1-5x minimal erythema dose) control- and antioxidant-treated skin using a solar simulator with a 295-nm band-pass filter. On day 5, we measured antioxidant protection factor, erythema, sunburn cells, and thymine dimers. RESULTS: The combination of 15% L-ascorbic acid and 1% alpha-tocopherol provided significant protection against erythema and sunburn cell formation; either L-ascorbic acid or 1% alpha-tocopherol alone also was protective but the combination was superior. Application during 4 days provided progressive protection that yielded an antioxidant protection factor of 4-fold. In addition, the combination of vitamins C and E provided protection against thymine dimer formation. CONCLUSION: Appreciable photoprotection can be obtained from the combination of topical vitamins C and E. We suggest that these natural products may protect against skin cancer and photoaging.     

Topical melatonin in combination with vitamins E and C protects skin from ultraviolet-induced erythema: a human study in vivo

In this randomized, double-blind human study, the short-term photoprotective effects of different antioxidants and their combinations were evaluated in vivo . Vitamin C (ascorbic acid), vitamin E (α-tocopherol) and melatonin ( N -acetyl-5-methoxytryptamine) were topically applied, alone or in combination, 30 min before ultraviolet-irradiation of the skin. The erythemal reaction was evaluated visually and non-invasively using different bioengineering methods (skin colour and skin blood flow). The results showed a modest protective effect of the vitamins when applied alone and a dose-dependent photoprotective effect of melatonin. Topical application of combinations of both vitamins, or of melatonin with vitamins, enhanced the photoprotective response. Better protection was obtained by using the combination of melatonin with both vitamins. The role of reactive oxygen species and oxygen-derived free radicals, as well as potential sunscreening properties of the employed antioxidants, are discussed in view of possible mechanisms to explain this elevated photoprotective effect.     

Photoprotective effect of topical anti-inflammatory agents against ultraviolet radiation-induced chronic skin damage in the hairless mouse

Albino hairless mice (Skh:HR-1) exposed chronically to suberythemal doses of ultraviolet (UV) radiation display visible and histological alterations in the skin. One alteration is an increase in dermal cellularity, including inflammatory cells. This suggested a role for inflammation in chronic photodamage. We evaluated the photoprotective effect of topical hydrocortisone, ibuprofen, and naproxen against photodamage. All 3 agents protected against UVB radiation-induced visible wrinkling, tumor formation, and histological alterations. Hydrocortisone and naproxen were also evaluated for protection against UVA radiation-induced visible skin sagging and histological alterations. Both were very effective. These data indicate that chronic topical application of anti-inflammatory agents provides broad solar UV spectrum photoprotection.     

Cutaneous vitamins A and E in the context of ultraviolet- or chemically-induced oxidative stress

Vitamins A and E are present in mammalian skin. Although the main circulating form of vitamin A in the blood is retinol, the epidermis stores it as retinyl esters. The epidermis can be easily loaded with high amounts of vitamin A by topical application of either retinol or retinaldehyde, two well-tolerated precursors of the biologically active retinoic acid, while topical alpha-tocopherol loads the epidermis with vitamin E. The probable physiological function of epidermal vitamin E is to contribute to the antioxidant defense of the skin, whereas that of epidermal vitamin A (retinol and retinyl esters) is not yet well understood. Besides being a precursor for retinoic acid, vitamin A also has a free radical scavenging potential. Due to their physical properties, vitamins A and E absorb ultraviolet (UV) light in the region of solar spectrum that is responsible for most of the deleterious biological effects of the sun. In the mouse, topical vitamin A has been shown to prevent the UV-induced epidermal hypovitaminosis A, while topical vitamin E prevents oxidative stress and cutaneous and systemic immunosuppression elicited by UV. Thus constitutive epidermal vitamins A and E appear complementary in preventing UV-induced deleterious cutaneous and systemic effects, and these properties can be reinforced by topical application of retinol or retinaldehyde and topical alpha-tocopherol.     

Novel nanocapsule of α‐lipoic acid reveals pigmentation improvement: α‐Lipoic acid stimulates the proliferation and differentiation of keratinocyte in murine skin by topical application

α‐Lipoic acid is amphipathic with low molecular sulphur‐containing fatty acid and has strong antioxidant effects. It has been used at the purposes of anti‐ageing, treatment of diabetic neuropathy, and supplement as antioxidant. Though α‐lipoic acid is normally administered in oral or injection, it has not been used in a topical use via skin because of its bad penetration. We developed the novel nanocapsule of α‐lipoic acid, named α‐lipoactive (nLA), to improve skin permeability. The nLA is constructed as micelles of α‐lipoic acid mixed with the non‐ionic surfactant, and its surface of the micelles was coated with inorganic metal salts. It is water soluble and has a diameter of approximately 8‐15 nm. After nLA was applied to the murine skin, epidermal thickening was observed. It was confirmed that this effect is caused by α‐lipoic acid molecule, but not by the raw material used for encapsulation. In in vivo experiments, it was found that nLA is very effective for improving UV‐induced pigmentation and epidermal thickening. Our findings suggest that nanoencapsulation of α‐lipoic acid is considerably effective for topical application.     

Functional regulation of tyrosinase and LAMP gene family of melanogenesis and cell death in immortal murine melanocytes after repeated exposure to ultraviolet B

The aim of this study was to investigate the effects of topical α-tocopherol application on epidermal and dermal antioxidants and its ability to prevent ultraviolet (UV)-induced oxidative damage. Hairless mice received topical applications of α-tocopherol 24 h before a single, acute UV irradiation (10 × minimal erythemal dose). The four major antioxidant enzymes (catalase, superoxide dismutase, glutathione reductase and glutathione peroxidase), hydrophilic and lipophilic antioxidants, and lipid hydroperoxides, markers of oxidative damage, were assayed in both epidermis and dermis of hairless mice. Topical α-tocopherol treatment increased dermal superoxide dismutase activity by 30% ( P  < 0.01) and protected epidermal glutathione peroxidase and superoxide dismutase from depletion after UV irradiation. Total and reduced glutathione levels in the epidermis increased by 50% after the topical treatment ( P  < 0.05), as did dermal ascorbate levels (by 40%; P  < 0.01). The topical treatment increased α-tocopherol levels both in the epidermis (62-fold) and the dermis (22-fold; P  < 0.001 in each layer). Furthermore, α-tocopherol treatment significantly reduced the formation of epidermal lipid hydroperoxides after UV irradiation ( P  < 0.05). These results demonstrate that topical administration of α-tocopherol protects cutaneous tissues against oxidative damage induced by UV irradiation in vivo , and suggest that the underlying mechanism of this effect involves the up-regulation of a network of enzymatic and non-enzymatic antioxidants.     

Skin photoprotection by natural polyphenols: anti-inflammatory, antioxidant and DNA repair mechanisms

Epidemiological, clinical and laboratory studies have implicated solar ultraviolet (UV) radiation in various skin diseases including, premature aging of the skin and melanoma and non-melanoma skin cancers. Chronic UV radiation exposure-induced skin diseases or skin disorders are caused by the excessive induction of inflammation, oxidative stress and DNA damage, etc. The use of chemopreventive agents, such as plant polyphenols, to inhibit these events in UV-exposed skin is gaining attention. Chemoprevention refers to the use of agents that can inhibit, reverse or retard the process of these harmful events in the UV-exposed skin. A wide variety of polyphenols or phytochemicals, most of which are dietary supplements, have been reported to possess substantial skin photoprotective effects. This review article summarizes the photoprotective effects of some selected polyphenols, such as green tea polyphenols, grape seed proanthocyanidins, resveratrol, silymarin and genistein, on UV-induced skin inflammation, oxidative stress and DNA damage, etc., with a focus on mechanisms underlying the photoprotective effects of these polyphenols. The laboratory studies conducted in animal models suggest that these polyphenols have the ability to protect the skin from the adverse effects of UV radiation, including the risk of skin cancers. It is suggested that polyphenols may favorably supplement sunscreens protection, and may be useful for skin diseases associated with solar UV radiation-induced inflammation, oxidative stress and DNA damage.     

Infrared radiation-induced matrix metalloproteinase in human skin: implications for protection

Human skin is exposed to infrared radiation (IR) from natural and artificial sources. In previous studies, near IR radiation (IRA; 760-1,440 nm) was shown to elicit a retrograde mitochondrial signaling response leading to induction of matrix metalloproteinase-1 (MMP-1) expression. These studies, however, have exclusively employed cultured human skin fibroblasts ex vivo. Here, we have assessed the in vivo relevance of these observations by exposing healthy human skin in vivo to physiologically relevant doses of IRA. Eighty percent of the tested individuals responded to IRA radiation by upregulating of MMP-1 expression. Specifically, IRA irradiation caused increased expression of MMP-1 in the dermis, but not in the epidermis. Raman spectroscopy revealed that IRA radiation also caused a significant decrease in the antioxidant content of human skin. In vitro studies had previously shown that IRA-induced MMP-1 expression was mediated through an oxidative stress response, which originates from the mitochondrial electron transport chain. We now report that incubation of cultured human dermal fibroblasts or treatment of human skin with specific antioxidants prevented IRA radiation-induced MMP-1 expression in vitro and in vivo. Thus, IRA irradiation most likely promotes premature skin aging and topical application of appropriate antioxidants represents an effective photoprotective strategy.     

Topical treatment of skin aging

Safe levels of UV exposure and UV protection are the most important measures to protect the skin from epithelial skin cancer and skin aging. This report reviews noninvasive topical methods to counteract skin wrinkling and irregular pigmentation of aging skin. Furthermore, information is provided about the effects of UV protection by using sunscreens and topical antioxidants. The effect of vitamin A acid derivatives, chemical peeling, and bleaching agents is considered. Newly developed substances are presented.     

The role of antioxidants in photoprotection: A critical review

Free radicals have long been studied as a contributor to aging and disease processes. Endogenous production of radicals from cellular metabolism and exogenous sources from ultraviolet radiation and pollution can damage the skin on the cellular and tissue levels. Although the body possesses an elegant defense system to prevent radical damage, this innate system can be overwhelmed and lead to a state of oxidative stress or immunosuppression, and can even trigger carcinogenesis. Topical supplementation of antioxidants can provide additional protection to neutralize reactive oxygen species from both endogenous and exogenous sources. This review will discuss our current understanding of the mechanisms of free radical damage and evaluate the potential benefit of topical antioxidants in sunscreens and skin care products.     

Ferulic acid stabilizes a solution of vitamins C and E and doubles its photoprotection of skin

Ferulic acid is a potent ubiquitous plant antioxidant. Its incorporation into a topical solution of 15%l-ascorbic acid and 1%alpha-tocopherol improved chemical stability of the vitamins (C+E) and doubled photoprotection to solar-simulated irradiation of skin from 4-fold to approximately 8-fold as measured by both erythema and sunburn cell formation. Inhibition of apoptosis was associated with reduced induction of caspase-3 and caspase-7. This antioxidant formulation efficiently reduced thymine dimer formation. This combination of pure natural low molecular weight antioxidants provides meaningful synergistic protection against oxidative stress in skin and should be useful for protection against photoaging and skin cancer.     

In vivo and in vitro evaluation of the use of a newly developed melatonin loaded emulsion combined with UV filters as a protective agent against skin irradiation

BackgroundMelatonin has attracted attention because of their high antioxidant and anticarcinogenic activity. Otherwise, the use of sunscreens is recommended for patients after chemotherapy and radiotherapy treatments or to prevent UV radiation-induced skin damages that may result in pre-cancerous and cancerous skin lesions.ObjectiveTo evaluate the beneficial influence of melatonin in topical sunscreen emulsions combined with three common ultraviolet filters.MethodsAfter the formulation characterization in terms of rheology, stability studies were performed. Release studies let us to evaluate its mechanism of delivery and ex vivo permeation study through human skin, the amount of melatonin retained. The antioxidant activity assay was also carried out, and finally the in vivo photoprotective effect in rats was tested as transepidermal water loss and erythema formation.ResultsThe rheological behaviour of formulations was pseudoplastic fluid, all emulsions had good physical stability. Release studies showed a trend of enhancement in melatonin release from emulsions incorporating UV filters and followed a Weibull model. Melatonin permeation was higher from the emulsion containing melatonin combined with a mixture of three ultraviolet filters (MMIX) formulation. Equally this formulation exhibited the highest radical scavenging activity. Finally the photoprotective assay showed that only skin areas treated with this formulation were statistically equivalent to the unirradiated control area.ConclusionMMIX formulation would be a promising formulation for preventing the undesirable adverse effects of UV skin irradiation because melatonin not only acts as a potent antioxidant itself, but also is capable of activating an endogenous enzymatic protective system against oxidative stress.     

Topical indomethacin protects from UVB and UVA irradiation

Conclusions Indomethacin is known to decrease the UVB (290–310 nm) induced erythema when applied topically in regular intervals after irradiation [2,5,6], probably via inhibition of prostaglandin synthesis. The absorption spectrum of an IM solution (Fig. 1) suggests that this drug may also exert a high filter capacity in the UVB and UVA range, when applied as a topical suncreen before exposure to UV light. The present study demonstrates that a 2.5% solution of IM indeed protects from UVB induced erythema, and these results are in accordance with those of Lim et al., recently published in abstracted form (1983). The present study demonstrates that topically aplied IM exerts a photoprotective effect also at the cellular level since it prevents UV induced keratinocyte cell death (formation of sunburn cells).Topical application of 2.5% IM decreases the induction of PUVA erythema and inhibits the development of immediate pigment darkening, which demonstrates a potent photoprotective effect also in the UVA range.The photoprotective mechanism of IM when applied prior to irradiation seems to be a filter effect (as to be expected from the absorption spectrum) rather than a biochemical action such as inhibition of prostaglandin synthesis, although the latter explanation cannot be excluded completely from the present results. Preliminary data, however, indicate that sunburn cells do occur in skin irradiated with erythemogenic doses of UVB light and treated with topical IM after exposure, which prevents erythema formation. Before IM can be widely used, it is necessary to evaluate optimal concentration and vehicles and in particular percutaneous absorption and possible systemic toxicity. The UVB screening potency of topical IM seems to be similar to other sunscreens (e.g. para-aminobenzoic acid/PABA), but IM offers a much higher protection against UVA light. This may be important in the treatment of certain photodermatoses and also in the prevention of elastotic skin damage by sunlight.     

Effect of topical antioxidants on UV-induced erythema formation when administered after exposure

BACKGROUND: Photoprotective effects of topically applied antioxidants when applied before ultraviolet radiation (UVR) exposure are well known. Their protective effect when applied after UVR exposure is, however, less established. OBJECTIVE: In a randomized, double-blinded, placebo-controlled human study the short-term photoprotective effects of different antioxidants and of their combinations were evaluated when applied after UVR exposure. METHODS: Melatonin (N-acetyl-5-methoxytryptamine), vitamin E (alpha-tocopherol) and vitamin C (ascorbic acid) were topically administered alone or in combination following UVR exposure as single applications (immediately or 30 min after irradiation, respectively) or as multiple applications (three times: 30 min, 1 h and 2 h after irradiation). The erythemal reaction was evaluated visually and noninvasively with bioengineering methods (skin color and skin blood flow). RESULTS: No significant protective effect of melatonin or the vitamins when applied alone or in combination were obtained when antioxidants were applied after UVR exposure. No improved photoprotective effect was obtained when multiple applications were done. CONCLUSION: UVR-induced skin damage is a rapid event, and antioxidants possibly prevent such damage only when present in relevant concentration at the site of action beginning and during oxidative stress.     

Phototoxicity to diuretics and antidiabetics in the cultured keratinocyte cell line HaCaT: evaluation by clonogenic assay and single cell gel electrophoresis Comet assay)

BACKGROUND: Potential phototoxicity has been described for a number of drugs and chemical substances. Psoralens, chlorpromazines and fluoroquinolones have been described as inducing photomutagenicity and photocarcinogenicity in vitro and in vivo. We wanted to investigate oral antidiabetics and diuretics for potential phototoxicity and possible DNA damage in the HaCaT cell line. METHODS: : The oral antidiabetics tolbutamide, glibenclamide and glipizide, and the diuretics bendroflumethiazide, butizide, furosemide, hydrochlorothiazide and trichlormethiazide were dissolved in DMSO to final concentrations of 1 mM, 0.1 mM, and 0.01 mM, incubated together with the cells, and exposed to UVA1 (23 or 48 J/cm2). Cell survival was evaluated in a clonogenic assay and phototoxic DNA damage was investigated by single cell gel electrophoresis (comet assay). To investigate possible inhibiting effects of antioxidants, L-ascorbic acid and alpha-tocopherol were added at a final concentration of 1 mM 24 h before treatment with the drugs. RESULTS: Bendroflumethiazide, furosemide, hydrochlorothiazide, trichlormethiazide and tolbutamide induced dose-dependent phototoxicity in the clonogenic assay. Cells incubated with bendroflumethiazide, tolbutamide and glibenclamide and irradiated with UVA1 demonstrated increased oxidative DNA damage, revealed as alkali-labile sites in the comet assay. Pretreatment with L-ascorbic acid or alpha-tocopherol suppressed the UVA-induced DNA damage in cells incubated with 1 mM bendroflumethiazide, furosemide, glibenclamide, glipizide, tolbutamide or trichloromethiazide. CONCLUSION: Several oral antidiabetics and diuretics show phototoxic effects in the HaCaT cell line. Inhibiting effects of antioxidants point towards involvement of reactive oxygen species in phototoxic DNA damage, suggesting a link between the phototoxic and photocancerogenic potential of the sulfonamide-derived oral antidiabetic and diuretic drugs. Excessive exposure to UV light may be deleterious for patients treated with oral antidiabetic and diuretic drugs.     

Photoprotection of UV-irradiated human skin: an antioxidative combination of vitamins E and C,carotenoids, selenium and proanthocyanidins

Endogenous antioxidants are decreased in skin and blood during UV exposure. Combined supplementation of beta-carotene, alpha-tocopherol and ascorbic acid in addition to topical sunscreens may help to lower the risk of sunburning. Acute UV erythema with sunburn reaction are the most important factors in conjunction with the cumulative life-long UV dose for inducing skin damage resulting in photoageing and precancerous and cancerous lesions. Therefore, a clinical, randomized, double-blind, parallel group, placebo-controlled study was conducted in healthy young female volunteers (skin type II) investigating the preventive, photoprotective effect of supplementation with Seresis, an antioxidative combination containing both lipid and water-soluble compounds: carotenoids (beta-carotene and lycopene), vitamins C and E, selenium and proanthocyanidins. In this study, the oral administration of Seresis appeared to be well tolerated. The preparation contains antioxidant compounds in quantities occurring at physiological levels and can therefore be used safely over a long period of time. Despite the fact that the assessment of the light sensitivity (minimal erythemal dose, chromametry) of the skin did not show any statistically significant differences between the Seresis and the placebo group, a clear statistical trend, however, could be demonstrated, i.e. Seresis was able to slow down the time of the development and grade of UVB-induced erythema. The primary efficacy parameter matrix metalloproteinases 1 (MMP-1) between treatment and placebo group following UV irradiation showed a significant difference (p < 0.05), which occurred due to the fact that after a 2-week UV irradiation, MMP-1 slightly increased (p < 0.03) in the placebo group and decreased (p < 0.044) in the treated group. The MMP-9 changes showed a clear tendency of decrease in the Seresis group (p < 1.393) and increase (p < 0.048) in the placebo group. These data emphasise that supplementation with Seresis decreases the UV-induced expression of MMP-1 and 9, which might be important in photoprotective processes. From our data, we thus finally draw the conclusion that by the combination of antioxidants, such as in the formulation of Seresis, a selective protection of the skin against irradiation can be achieved. This might be important for future recommendations for immediate suppression of the early phase of UV-induced erythema, that means pharmacological prevention of sunburn reaction as well as subsequent chronic skin damage.     

Effects of low-dose ultraviolet radiation on in vivo human cutaneous recall responses

Relatively few studies have examined the effects of low-dose ultraviolet (UV) radiation on in vivo human cutaneous immunity, or the ability of sunscreens to prevent UV-induced immunosuppression. We have studied the effects of solar-simulated UV radiation on nickel contact hypersensitivity (CHS) in nickel-allergic volunteers, and on delayed type hypersensitivity responses in Mantoux-positive volunteers. Nickel CHS and Mantoux responses were significantly suppressed by acute, suberythemal UV exposures equivalent to less than 8 min summer sunlight. Both UVA and UVB wavebands were immunosuppressive, but UVA-induced immunosuppression was transient, whereas UVB had a more sustained effect. Dose-responses for UV immunosuppression were determined using the nickel method, enabling calculation of in vivo sunscreen immune protection factors in a manner analogous with sun protection factor measurement. Sunscreens were found to confer significantly less protection against UV-induced immunosuppression than against UV-induced erythema.