Measurement, analysis and prediction of topical UV filter bioavailability.

The aim of the present study was to objectively quantify and predict bioavailability of three sunscreen agents (i.e., benzophenone-3, 2-ethylhexylsalicylate, and 2 ethylhexyl-4-methoxycinnamate) in epidermis treated by petrolatum and emulsion-based formulations for 7 and 30min on four human volunteers. Profiles of sunscreen agents through stratum corneum (SC), derived from the assessment of chemical amounts in SC layers collected by successive adhesive tape-stripping, were successfully fitted to Fick's second law of diffusion. Therefore, permeability coefficients of sunscreen agents were found lower with petrolatum than with emulsion based formulations confirming the crucial role of vehicle in topical delivery. Furthermore, the robustness of that methodology was confirmed by the linear relationship between the chemical absorption measured after 30min and that predicted from the 7-min exposure experiment. Interestingly, in this dermatopharmacokinetic method, the deconvolution of permeability coefficients in their respective partition coefficients and absorption constants allowed a better understanding of vehicle effects upon topical bioavailability mechanisms and bioequivalence of skin products.

Skin penetration and sun protection factor of five UV filters: effect of the vehicle.

To gain information about efficacy and safety of sunscreens, we compared the skin penetration of ultraviolet (UV) filters from two vehicles, i.e. an oil-in-water (O/W) emulsion gel and petrolatum jelly both in vitro and in vivo, as well as the corresponding pharmacological effect, i.e. the sun protection factor (SPF) in vivo. The UV filters studied were benzophenone-3 (BPH), ethylhexyl methoxycinnamate (EHM), butyl methoxydibenzoyl methane, ethylhexyl salicylate and homosalate. The human skin penetration of these five chemicals from the two vehicles was determined both in vitro using Franz cells and in vivo using a standardized tape-stripping method. The SPF of the two sunscreens was determined in vivo following the COLIPA guidelines. In vitro none of the filters permeated through the skin after 6 h of product application and very little could be found in the skin. BPH and EHM were the only UV filters found in the dermis (both after 30 min and 6 h). An effect of the vehicle could be noticed only for BPH after 30 min in the dermis and 6 h in both dermis and epidermis. In vivo, no differences in the amount of individual UV filters (in % of the applied dose) in the 15 first strips of the stratum corneum (SC) were found following 30 min of application of the formulations; however, the amount of UV filters that were retained in the SC was significantly higher (around 3 times) with the O/W emulsion gel than with the petrolatum jelly. This difference between the two vehicles was also of consequence for the SPF in vivo measured 30 min after application of the products (SPF congruent with 18 with the O/W emulsion gel compared to SPF congruent with 10 with the petrolatum jelly). By choosing the right vehicle or optimizing it, not only sunscreen products can be significantly improved in terms of pharmacological efficacy but the potential toxicological risk associated with the skin penetration of UV filters may be significantly reduced.

Lip sun protection factor of a lipstick sunscreen.

BACKGROUND AND OBJECTIVE: There is a well-documented need for effective human UVA and UVB photoprotection. Since there are important anatomical variations, the sun protection factor (SPF) of a lipstick sunscreen was measured on the anatomical site intended for use. METHODS: The SPF tests were performed according to Federal US and European COLIPA guidelines. Prior to performing a test on the lip, the minimal erythemal dose (MED) of the unprotected back skin was determined. Subsequently, the sunscreen SPF was measured on the anatomical target site (lip). The evaluator was blinded with respect to scoring the SPF of each sunscreen treatment. Individual test sites were assigned to one of the following treatment conditions: (1) no treatment (MED of unprotected skin); (2) test formulation; (3) reference standard. RESULTS: The MED on unprotected back skin was found to be 25% lower than on unprotected lip skin. The SPF of the lipstick sunscreen was measured 2 units lower than the SPF determined in the classical way on the back skin. CONCLUSION: It was hypothesized that the higher MED of the lower lip compared with back skin was due to the adaptation of this tissue to the continuous exposure to UV radiation.

Microbiological quality of topical drug formulations: efficacy of antimicrobial preservation against Paecilomyces lilacinus.

BACKGROUND: Microbiological quality of topical products comprises both, the microbiological purity of the unopened product and the efficacy of the antimicrobial preservation system. OBJECTIVE: Subsequent to an outbreak of invasive Paecilomyces lilacinus mycosis among patients of an isolation ward, probably caused by a contaminated skin care product, the microbiological quality of different skin care products from the market was investigated. METHODS: The different products were investigated for their efficacy of antimicrobial preservation in general and especially against P. lilacinus according to a pharmacopoeial routine method slightly adopted for the purpose of this investigation. RESULTS: The products did partially not comply with the British Pharmacopoeia 1993 test for efficacy of antimicrobial preservation. The antimicrobial preservation systems were less effective against P. lilacinus than against the pharmacopoeial reference germs. The antimicrobial preservation efficacy decreased towards the end of the shelf-life of the product. A decreased P. lilacinus inoculum dose was related to an increased growth of the micro-organisms. CONCLUSION: Topical products are, unless not labelled otherwise, non-sterile preparations and their preservation systems are only tested against pharmacopoeial key micro-organisms. The microbiological behaviour following contamination with other germs remains unknown.

Photostabilization of butyl methoxydibenzoylmethane (Avobenzone) and ethylhexyl methoxycinnamate by bis-ethylhexyloxyphenol methoxyphenyl triazine (Tinosorb S), a new UV broadband filter.

It is now well documented that chronic UVA exposure induces damage to human skin. Therefore, modern sunscreens should not only provide protection from both UVB and UVA radiation but also maintain this protection during the entire period of exposure to the sun. UVA filters, however, are rare and not sufficiently photostable. We investigated the effect of the introduction of a new UV filter, bis-ethylhexyloxyphenol methoxyphenyl triazine (Tinosorb S), in oil in water sunscreen formulations on the photostability of butyl methoxydibenzoylmethane (Avobenzone [AVB]) after irradiation with an optically filtered Xenon arc source (UV irradiance adjusted at 1 mean effective dose [MED]/min). With spectrophotometrical methods to assess the sun protection factor (SPF) and UVA ratio and chromatographical methods to determine the amount of UV filters recovered after irradiation we showed that Tinosorb S prevented the photodegradation of AVB in a concentration-dependent way, leading to a sustained SPF and UVA ratio even after irradiation with doses of up to 30 MED. Since AVB was shown to destabilize ethylhexyl methoxycinnamate (EHM) we tested the effect of Tinosorb S in sunscreens containing this UV filter combination. Here too Tinosorb S showed photoprotective properties toward both UV filters. Thus, Tinosorb S can be used successfully to improve the photostability and efficiency of sunscreens containing AVB and EHM.

There is no influence of a temperature rise on in vivo adsorption of UV filters into the stratum corneum.

Temperature influences the stratum corneum adsorption of several topically applied compounds. This study was designed to evaluate the influence of the temperature on the stratum corneum adsorption of 3 UV filters. The UV filters were solubilized in two vehicles, an emulsion gel and petroleum jelly and applied at respectively, 31 and 40 degrees C during 30 min. In vivo stratum corneum UV filter content was measured using the tape stripping method. Similar amounts of UV filter were detected in the stratum corneum when comparing applications at the different temperatures. Application of the UV filters in the emulsion gel resulted in higher stratum corneum UV filter concentrations compared with application in the petroleum jelly. The application temperature did not influence the stratum corneum adsorption of the tested UV filters while the nature of the vehicle significantly influenced the amount of UV filters recovered from the stratum corneum.

Surfactant irritation: in vitro corneosurfametry and in vivo bioengineering.

BACKGROUND/AIMS: Irritant reactions to surfactants, cleansing products, soaps and detergents are common in clinical and occupational dermatology. Mildness has become a major benefit claimed, and testing for mildness now ranks among the first concerns of the manufacturing industry. A wealth of publications deals with this problem, trying to improve the methodology, reduce the costs of testing and facilitate decision-making. Differences in vivo can be measured clinically and/or instrumentally. This is difficult, as commercially available products are generally safe to use and none are harsh in the absolute sense. METHODS: Nineteen different products (syndets, shampoos, personal cleansers), all claiming to be mild, were tested in vitro by a newly introduced method, corneosurfametry. For evaluating the aggressiveness of the products, the calculation of an index of irritation (IOI) was proposed. A concentration-effect curve of sodium lauryl sulfate (SLS) as standard and model surfactant was obtained. Some of the products were further tested in vivo with a flex wash test and with a soap chamber test and compared to SLS. Bioengineering methods (transepidermal water loss TEWL, skin color) were used to evaluate the results. RESULTS AND CONCLUSIONS: The results of the corneosurfametry allowed us to classify the products in three categories, with increasing aggressiveness towards the stratum corneum, according to their IOIs. The in vivo tests were not able to discriminate between the products, but ranks from the results of the bioengineering measurements showed a good correlation between TEWL changes, but not between colour changes, and IOIs from corneosurfametry. Corneosurfametry emerged as a simple, low-cost and fast method for ranking commercial products according to their mildness. However, the skin bioengineering techniques showed that some products could lead to skin reactions, such as erythema, that could not be detected by the in vitro technique.

Efficacy of a barrier cream and its vehicle as protective measures against occupational irritant contact dermatitis.

The actual advantage of barrier creams over bland emollients for skin protection is still hotly debated. In a randomized, double-blinded study, a newly-introduced barrier cream and its moisturizing vehicle were compared regarding their skin compatibility, efficacy and resulting acceptance. Thus, 2 panels of 25 hospital nurses with mild signs of skin irritation were asked to use 1 of the test products provided (verum or vehicle) over a period of 4 weeks. Effects of both types of preparations were studied weekly by clinical examination and the instrumental assessment of bioengineering parameters. Results showed no significant differences between barrier cream and vehicle. In both groups, clinical skin status improved and stratum corneum hydration increased significantly during the study period. Both preparations were tolerated and accepted well, thus showing both skin protection and skin care. These results contribute to the debate as to whether a strict distinction between "skin care" and "skin protection" products is justified. The vehicle alone is capable of positively influencing skin status. Emphasis must be laid on regular, frequent, and correct application of a product for it to be effective.

A qualitative estimate of the influence of halcinonide concentration and urea on the reservoir formation in the stratum corneum.

The existence of a stratum corneum reservoir for topically applied substances is well known. Data concerning the stratum corneum retention time are important for the elaboration of optimal topical treatment. We used the re-occlusion technique followed by skin colour measurements (Chromametry) for the evaluation of the stratum corneum retention time of halcinonide. We found a significant reservoir for halcinonide up to 5 days after the initial application. The retention was found to be corticosteroid concentration and formulation dependent.

Percutaneous absorption of salicylic acid in man after topical administration of three different formulations.

OBJECTIVE: To determine the amount of drug which is absorbed during 1 day following topical application of three different preparations containing salicylic acid. METHODS: Ten grams of the formulations, either (a) Kerasaltrade mark 5% ointment, (b) salicylic acid 5% or (c) 10% in petrolatum, were administered consecutively to a 600-cm2 area on alternating sides of the back of healthy volunteers (n = 9). Thirty minutes after application, a skin area of 2.54 cm2 was stripped with D-Squametrade mark adhesive disks to determine the amount of salicylic acid in the stratum corneum. The entire application site was then covered by a thin gauze bandage and was not washed for the next 24 h. Urine was collected for 26 h following administration, hydrolyzed and assayed by HPLC analysis. RESULTS: The absolute amounts absorbed and excreted were 52.6 +/- 29.4 mg (mean +/- SD), 127.1 +/- 43.9 mg and 208.0 +/- 81.7 mg, and the doses absorbed in relation to the doses applied (500 mg salicylic acid in case of formulations a and b and 1,000 mg for formulation c) were 9.3 +/- 3.8, 25.1 +/- 8.5 and 20.2 +/- 7.7%, respectively. The amounts of salicylic acid in the skin 30 min after application were 36.3 +/- 16.5, 18.2 +/- 11.9 and 31.3 +/- 15.4 microg/ cm2 as determined by the tape stripping procedure. CONCLUSIONS: Significant differences in the doses absorbed were detected between the two formulations a and b (same concentration) with different vehicles (p value < 0.001) as well as between b and c (same vehicle) with different concentrations (p value = 0.018) using Student's paired t test. These results demonstrate that salicylic acid is well absorbed by healthy skin.

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.

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 (alpha-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.

Measurement of sodium lauryl sulfate-induced skin irritation.

Besides visual evaluation, skin irritation induced by sodium lauryl sulfate (SLS) may be characterized by bioengineering measurements, such as skin colour reflectance, transepidermal water loss (TEWL) or hydration. Short application times or low concentrations of the irritant usually do not modify the visual aspect of the skin, and the measurements described above are unchanged or only slightly altered. We were looking for a suitable method to measure cutaneous changes not detectable by usual bioengineering procedures. Therefore these measurements were compared to those of dynamic function testing of the stratum corneum, namely sorption-desorption and moisture accumulation tests. Different concentrations of SLS (0.1%, 0.5%, 2.5%), application times (15 min, 24 h) and times of testing (1 h, 24 h after patch removal) were investigated on the ventral forearm of human subjects. When SLS was applied for a short period (15 min), 1 h after patch removal skin colour, TEWL and hydration were not modified, while increases in hygroscopicity, water-holding capacity and water accumulation were detected depending on the applied concentration. Increase of hygroscopicity was closely correlated with the alteration of epidermal barrier function (TEWL). We demonstrated that sorption-desorption and moisture accumulation tests performed on SLS-treated areas for a short period, without visible modifications, could evaluate changes of the stratum corneum properties. We consider these tests as useful complementary methods to skin colour, TEWL and hydration measurements, particularly in the detection of subclinical skin injuries.

Skin penetration and sun protection factor of ultra-violet filters from two vehicles.

PURPOSE: In order to improve our knowledge on the efficacy and safety of sunscreen products, we measured the skin penetration profiles of ultra-violet (UV) filters in vitro and in vivo, and the corresponding sun protection factors (SPF) from two vehicles (an O/W emulsion-gel and petroleum jelly). METHODS: The UV filters tested were oxybenzone (5%, A), 2-ethylhexyl 4-methoxycinnamate (7.5%,B), and 2-ethylhexylsalicylate (3%,C). Two mg/cm(2) were applied for 2 min to 6 h. In vitro penetration measurements were performed with static diffusion cells. In vivo, horny layer concentrations were measured after stripping and the SPF evaluated as recommended by the COLIPA-guidelines. RESULTS: Significant differences between vehicles were noticed in vitro as well as in vivo. In vitro, the emulsion-gel generated higher epidermal concentrations than petroleum jelly. Values at 6 h, expressed as percent of the applied dose for A, B, and C were 4, 9, and 7% for the emulsion-gel and 2, 1, and 2% for petroleum jelly. An opposite trend was noticed, mainly for A, in the deeper skin layers with concentrations of 2% in the dermis and 5% in the receptor fluid for petroleum jelly and 0.6% and 1% for the emulsion-gel respectively. In vivo, for each UV filter, maximal stratum corneum levels (15 strips) were obtained at 0.5 h with percentages of the applied doses of 50% for the emulsion-gel and 15 percent for petroleum jelly. SPFs, measured 0.5 h after application amounted to 14 for the emulsion-gel and 5 for petroleum jelly, and decreased in both cases by a factor 2.2 after removal of non penetrated product. CONCLUSIONS: These preliminary results demonstrated that UV filters penetration and retention as well as expected SPF could be optimized by a suitable vehicle.

[Helicobacter pylori eradication therapy with bismuth citrate/amoxycillin combination therapy]. / Helicobacter pylori-Eradikationstherapie mit einer Wismutzitrat/Amoxicillin-Kombinationstablette.

The efficacy of a new combination preparation containing bismuth citrate and amoxicillin in one tablet was compared with the efficacy of bismuth citrate monotherapy in a randomised double-blind study on the eradication of Helicobacter pylori. The study involved 70 H. pylori positive (antrum biopsies showing a positive urease test) patients with non-ulcer dyspepsia and chronic gastritis. The treatment period was 14 days; 35 patients in group 1 received 2 tablets tid containing the bismuth citrate amoxicillin combination (BIAM tablet; 250 mg amoxicillin base and 120 mg bismuth); 35 patients in group 2 were treated with 2 tablets tid containing bismuth citrate (BI tablet; 120 mg bismuth). Total daily dose was therefore 1500 mg amoxicillin + 720 mg bismuth in group 1 patients resp. 720 mg bismuth in group 2 patients. 4 weeks after therapy H. pylori could not be histologically detected in the antrum of 22 patients (63%) in group 1 and 8 patients (24%) in group 2. Thus in group 1 (BIAM) a significantly higher eradication rate (p < 0.001) was shown than in group 2 (BI). Inflammation characterized by the infiltration of polymorphonuclear cells was significantly (p < 0.01) less pronounced in group 1 (BIAM) than in group 2 (BI) 4 weeks after the end of treatment. Gastrointestinal distress was quantified by evaluation of 13 different symptoms using a fourpoints scale at the beginning of the study and after 2 and 6 weeks. The sum of scores decreased by 81% in group 1 (BIAM) and 71% in group 2 (BI) after 6 weeks.

Emollients and photo(chemo)therapy: a call for caution.

BACKGROUND: Emollients used for skin care, treatment of a disease or lubrication, before irradiation during the course of phototherapy or photochemotherapy, may significantly interfere with the transmission of UV light and may influence the outcome of the therapy. OBJECTIVE: These experiments should characterize and quantify the possible interactions of usual emollients with UV light transmission. METHOD: The method was an in vitro technique similar to the one already used for the determination of sun protection factors of topical sunscreen preparations. RESULTS: Among the products tested, two showed a significant photoprotection capable of interfering with the course of therapy. Oils enhanced UV light transmission significantly. CONCLUSIONS: Emollients may show significant interference with UV light transmission during phototherapy or photochemotherapy. The outcome and course of the therapy could thus be significantly modified.

Barrier function of the skin in a repetitive irritation model and influence of 2 different treatments.

BACKGROUND/AIMS: The aim of this study was to develop and characterize an experimental model of cumulative irritant contact dermatitis using the repetitive application over 2 weeks of 2 different irritative stimuli, sodium lauryl sulphate (SLS) in low concentration and toluene. Further, it was intended to look at the effect of 2 different treatments which could influence the regeneration of the skin barrier, as already investigated in preliminary short-term experiments. METHODS: Occlusive short-time daily application of the irritants (SLS 30 min; toluene 10 min) was performed. After removal of the occlusive patch, the irritated areas were treated with 2 different preparations, Lotio Alba Aquosa and vaseline (white petroleum jelly). Bioengineering techniques were used to precisely measure the skin condition. RESULTS: A mild irritation was obtained under these experimental conditions. The bioengineering measurements allowed us to differentiate both irritants through their different influences on trans-epidermal water loss and skin hydration. Treatment with vaseline did not significantly influence the course of the irritation, but Lotio Alba Aquosa clearly potentiated it, whereby different effects were noticed for both irritative stimuli. CONCLUSIONS: The use of bioengineering techniques in this model of mild cumulative irritation allowed us to differentiate the action of both irritants, showing that they act through different mechanisms. The results of the treatments pointed out the need to treat an irritation with the right product for the right time period.

Bioengineering measurements of barrier creams efficacy against toluene and NaOH in an in vivo single irritation test.

BACKGROUND/AIMS: By now, only a few models have been published with the goal of testing barrier creams in vivo in humans. The aim of this study was to evaluate with a single irritation test, barriers creams in humans against a lipophilic and a hydrophilic irritant, toluene and NaOH, respectively. METHODS: Both irritants were applied for 15 min after pretreatment of the skin with barrier creams. Non-invasive bioengineering methods, such as skin colorimetry (a*) and cutaneous blood flow (CBF) measurements were used to assess product protection. RESULTS: After toluene application on control sites, the irritation appeared quickly (Tmax =3 min after patch removal), was significant (+5-6 units for a* and+80% for CBF) and did not return to base value within 1 h. Skin irritation after NaOH application, as measured by a*, was less important (+2 units) and occurred later (T"max=40 min after patch removal). For this irritant, CBF response was minor and variable. When testing barrier properties of the products, none of them were able to prevent the skin erythema induced by toluene. Against NaOH, one barrier cream as well as petrolatum and a fatty cream protected the skin significantly. CONCLUSION: The present study points out the unsatisfactory effectiveness of several commercially available barrier creams claimed to protect against lipophilic or hydrophilic irritants.

Stratum corneum dynamic function measurements after moisturizer or irritant application.

Two simple tests were conducted which allowed the quantification of parameters that characterize the stratum corneum (SC) dynamic functions in vivo under physiological conditions after moisturizer applications for 1 h and after irritation with different concentrations of sodium lauryl sulphate (SLS; 0.5-4%) applied under occlusion for 15 min or 24 h. Both tests, the sorption-desorption test (SDT) and the moisture accumulation test (MAT), were performed with a Nova Dermal Phase Meter 9003. The following parameters were quantified: prehydration state (SDT, MAT), hygroscopicity, water-holding capacity (SDT), water accumulation velocity and water accumulation (MAT). These procedures allowed the demonstration of the water-holding effect of urea contained in moisturizers. Differences between the long and the short application time of SLS were characterized by differences in SC dynamic functions while the hydration state was not changed. An effect on transepidermal water loss (TEWL) was noted only after the long application time, although the MAT clearly showed dynamic parameters to be changed after 15 min of treatment. These tests were simple in practice and allowed the demonstration of functional modifications of the SC while other parameters remained unchanged. They gave insight into possible action mechanisms of urea and SLS in the SC.