RESUMEN
The sun protection factor (SPF) is related to the selected UV filters. The objective of this study was to evaluate and compare the rheological behavior and texture profile of two sunscreen formulations and to correlate these data with the obtained SPF values. Two formulations (F1 and F2) were developed with the same type and amount of UV filters - whereby one of them also contained ethoxylated lanolin as additional film former (F2). Their rheological behavior, texture profile and in vitro and in vivo SPF were analyzed. The film-forming properties were evaluated by skin profilometry and diffuse reflectance spectroscopy. The structures of the formulations were examined by two-photon tomography combined with fluorescence lifetime imaging, and the penetration profile into the stratum corneum was investigated by tape stripping. The formulation with lanolin presented lower and constant values for physical-mechanical parameters, with a higher and better reproducible SPF. Both formulations did not penetrate the viable epidermis. In conclusion, formulations with better surface deposition on the skin surface can influence the film formation and, consequently, improve the SPF. These findings are important to improve the efficacy of sunscreen formulations and reduce the addition of UV filters.
Asunto(s)
Factor de Protección Solar , Protectores Solares , Piel , Análisis Espectral , Rayos UltravioletaRESUMEN
BACKGROUND: The hair follicle represents a significant penetration route for topically applied substances. ISSUE: The percutaneous absorption of substances can be significantly increased and accelerated by the involvement of hair follicles. In addition, nanoparticles have the characteristic to penetrate deeply and effectively into the hair follicles. MATERIALS AND METHODS: An optimization of drug delivery for topically applied substances is possible if the nanoparticles act solely as a carrier to transport active ingredients into the hair follicle. Once the nanocarrier has penetrated into the hair follicle, the active substance must be released there. This can be triggered by various mechanisms. RESULTS: The released drug can thus pass into the living tissue surrounding the hair follicle independently. With the help of this innovative strategy, the bioavailability of topically applied substances can be significantly improved. CONCLUSION: The transport of active ingredients into the hair follicles with the help of particles and the release of active substances there is a very effective new method for transporting active substances through the skin barrier.
Asunto(s)
Sistemas de Liberación de Medicamentos , Folículo Piloso/metabolismo , Nanopartículas/metabolismo , Preparaciones Farmacéuticas/administración & dosificación , Absorción Cutánea/fisiología , Piel/metabolismo , Administración Cutánea , Materiales Biocompatibles , Transporte Biológico , HumanosRESUMEN
Sunlight is indispensable for life. Skin aging is determined not only by genetic factors but also by the influence of solar radiation which in all its spectral regions induces free radicals in the human skin. Below a critical concentration, free radicals are important for signaling processes in the human body. Exceeding a critical threshold value (FRTV), however, free radicals lead to cellular damage which, in turn, can induce skin aging, immunosuppression and even skin cancer. Recently it could be shown that 50% of the free radicals induced in the human skin by solar radiation were caused by the visible and infrared spectral regions. Taking into consideration that people who use sunscreen stay considerably longer in the sun than unprotected people, their skin could be exposed to free radical formation in the visible and infrared spectral regions, which far exceeds the critical threshold. Filter substances like those used in sunscreens for skin protection in the ultraviolet region are not available for the visible and infrared spectral regions. Using the natural protection mechanisms of the human skin as examples, however, the protective effect of sunscreens can be considerably enhanced by the addition of scattering and reflective pigments, such as titanium dioxide, as well as of antioxidants which neutralize the free radicals. Currently the focus of sunscreen development is shifting from mere UV protection to protection covering the entire solar spectrum.
Asunto(s)
Antioxidantes/administración & dosificación , Protección Radiológica/métodos , Enfermedades de la Piel/etiología , Enfermedades de la Piel/prevención & control , Luz Solar/efectos adversos , Protectores Solares/administración & dosificación , Titanio/administración & dosificación , Antioxidantes/química , Dermatología/tendencias , Medicina Basada en la Evidencia , Humanos , Protectores contra Radiación/administración & dosificación , Protectores contra Radiación/química , Energía Solar , Protectores Solares/química , Resultado del TratamientoRESUMEN
Sunscreens are a key pillar of the multimodal protection strategy against short- and long-term impacts of intermittent and continuous UV exposure. Hitherto, an unanswered part of current scientific discourse is the question whether a cosmetic pretreatment has an impact on distribution and adhesiveness of sunscreens on the skin and therefore affects UV protection. In order to evaluate the homogeneity of sunscreen filter distribution, water resistance as a parameter of adhesiveness and effective UV protection of sunscreens after a pretreatment with cream or lotion was investigated in 18 volunteers who were examined before and after swimming, using the established combination of the tape stripping procedure and UV/VIS spectroscopy. It was shown that a cosmetic skin pretreatment affects neither filter homogeneity nor effective UV protection prior to water contact. However, compared to nonpretreated skin, a considerable loss of water resistance is caused. Therefore, using a cream or lotion before application of sunscreens is not to be recommended.
Asunto(s)
Crema para la Piel/administración & dosificación , Protectores Solares/administración & dosificación , Adhesividad , Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Absorción Cutánea , Crema para la Piel/química , Protectores Solares/química , Agua/química , Adulto JovenRESUMEN
In this study, we compared the UV-protective abilities of the europium complex compared to titanium dioxide, which represents the most common physical filter for ultraviolet light in the broad-band spectral range. The UV absorption and light transformative capacities of the europium complex were evaluated using a spectrometer with a double-integrating sphere showing that the europium complex does not only absorb and reflect UV light, but transforms it into red and infrared light. It was found that the europium complex binds to the surface of Jurkat cells in vitro. Cells incubated with the europium complex showed a significantly higher viability after UVA and UVB irradiation as compared to untreated cells and cells incubated with titanium dioxide pointing out its photoprotective properties. The europium complex and titanium dioxide show similar penetration capacities into the stratum corneum as tested in human and porcine skin using tape stripping analysis. The europium complex has proved to be an efficient UV filter with a low cyto- and phototoxic profile and therefore represents a potential candidate for use in sunscreen formulations.
Asunto(s)
Europio/farmacología , Nanopartículas del Metal/administración & dosificación , Protectores Solares/farmacología , Titanio/farmacología , Adulto , Animales , Transporte Biológico , Permeabilidad de la Membrana Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Femenino , Humanos , Células Jurkat , Piel/metabolismo , Piel/efectos de la radiación , Absorción Cutánea , Porcinos , Rayos UltravioletaRESUMEN
The objective of the present investigation was to examine the utilization of optical and spectroscopic methods for the noninvasive characterization of Anthelios XL Fluide Extreme (SPF 50+), an exemplary sunscreen, concerning its homogeneity of distribution on the skin, its spectroscopic properties and its overall protective efficacy. The homogeneity of the distribution of the sunscreen on the skin was investigated with a multiphoton tomography microscope. Additionally, the sum transmission spectrum was determined using tape stripping and spectroscopic measurements. The results revealed a very homogeneous distribution of the sunscreen on the skin surface and also in the deep furrows. The sum transmission spectrum reflects a high protective efficacy of the sunscreen in both the UVA and UVB ranges. The sunscreen Anthelios XL Fluide Extreme (SPF 50+) generates a comfortable feeling on the skin and can be easily distributed. The presented optical methods have been shown to be suitable to investigate the overall protective efficacy of sunscreen products objectively, noninvasively and quickly.
Asunto(s)
Piel/metabolismo , Protectores Solares/farmacocinética , Adulto , Humanos , Persona de Mediana Edad , Sensación , Absorción Cutánea , Análisis Espectral , Tomografía , Resultado del TratamientoRESUMEN
The efficacy of a drug is characterized by its action mechanism and its ability to pass the skin barrier. In this article, different methods are discussed, which permit this penetration process to be analysed non-invasively. Providing qualitative and quantitative information, tape stripping is one of the oldest procedures for penetration studies. Although single cell layers of corneocytes are removed from the skin surface, this procedure is considered as non-invasive and is applicable exclusively to the stratum corneum. Recently, optical and spectroscopic methods have been used to investigate the penetration process. Fluorescence-labelled drugs can be easily detected in the skin by laser scanning microscopy. This method has the disadvantage that the dye labelling changes the molecular structures of the drug and consequently might influence the penetration properties. The penetration process of non-fluorescent substances can be analysed by Raman spectroscopy, electron paramagnetic resonance, CARS and multiphoton microscopic measurements. Using these methods, the concentration of the topically applied formulations in different depths of the stratum corneum can be detected by moving the laser focus from the skin surface deeper into the stratum corneum. The advantages and disadvantages of these methods will be discussed in this article.
Asunto(s)
Absorción Cutánea , Administración Tópica , Espectroscopía de Resonancia por Spin del Electrón , Colorantes Fluorescentes , Humanos , Microscopía/métodos , Espectrometría RamanRESUMEN
The inhomogeneous distribution of topically applied substances due to decisive differences in the skin structure (furrows and wrinkles) affects the efficacy of cosmetic products, in particular sunscreens. The combination of tape stripping and optical spectroscopy results in absorption data, which reflect ex vivo the inhomogeneity of the in vivo distribution of topically applied substances. Based on these data, a factor of inhomogeneity is defined describing the individual distribution of formulations on the skin surface of volunteers. Thus, the influence of different skin surface structures and the influence of different formulations on the distribution of the topically applied substances can be determined. Analyzing the inhomogeneity data on 6 volunteers (5 sunscreens per volunteer), it was found that the influence on the distribution of sunscreens caused by the formulation was higher than the inhomogeneity originating from the differences in the skin surface structure of the volunteers. The method is well suited to characterize, for example, sunscreens and antiaging creams in the process of development, as well as for the evaluation of the final products.
Asunto(s)
Absorción Cutánea , Piel/metabolismo , Protectores Solares/administración & dosificación , Protectores Solares/metabolismo , Administración Cutánea , Adulto , Química Farmacéutica , Emulsiones , Alemania , Humanos , Microscopía Confocal , Piel/anatomía & histología , Espectrofotometría Ultravioleta , Protectores Solares/química , Adulto JovenRESUMEN
Colonization and infection of wounds represent a major reason for the impairment of tissue repair. Recently, it has been reported that tissue-tolerable plasma (TTP) is highly efficient in the reduction of the bacterial load of the skin. In the present study, the antiseptic efficacy of TTP was compared to that of octenidine hydrochloride with 2-phenoxyethanol. Both antiseptic methods proved to be highly efficient. Cutaneous treatment of the skin with octenidine hydrochloride and 2-phenoxyethanol leads to a 99% elimination of the bacteria, and 74% elimination is achieved by TTP treatment. Technical challenges with an early prototype TTP device could be held responsible for the slightly reduced antiseptic properties of TTP, compared to a standard antiseptic solution, since the manual treatment of the skin surface with a small beam of the TTP device might have led to an incomplete coverage of the treated area.
Asunto(s)
Antiinfecciosos Locales/farmacología , Glicoles de Etileno/farmacología , Gases em Plasma/uso terapéutico , Piridinas/farmacología , Piel/efectos de los fármacos , Administración Cutánea , Adulto , Antiinfecciosos Locales/administración & dosificación , Antioxidantes/metabolismo , Antisepsia/métodos , Bacterias/efectos de los fármacos , Bacterias/aislamiento & purificación , Combinación de Medicamentos , Glicoles de Etileno/administración & dosificación , Femenino , Humanos , Iminas , Masculino , Piridinas/administración & dosificación , Piel/microbiologíaRESUMEN
Bacteria and fungi are located in the stratum corneum and the hair follicles. Therefore, the development and assessment of efficient drugs requires standard in vivo investigation methods permitting a differentiation between intercellular and follicular penetration and storage of topically applied anti-microbial substances. In the present study, the penetration and storage of Isoconazole nitrate in the stratum corneum and hair follicles was investigated by differential stripping after a 14-day topical application period and during a follow-up period of a further 21 days. One week after the application had terminated, Isoconazole nitrate could still be detected in concentrations above the minimal inhibition concentration in the stratum corneum and the hair follicles. In some subjects, Isoconazole nitrate could even be detected 14 days after the last application. No relevant changes in TEWL values were measured, indicating that the investigated compound did not induce an impairment of the barrier function. The study showed that differential stripping is suited to investigate the penetration and storage of topically applied substances into the stratum corneum and the hair follicles. Also, the hair follicles are a long-term reservoir for topically applied substances. This is of clinical importance, where a long-lasting therapeutic effect beyond the application time is required.
Asunto(s)
Antiinfecciosos Locales/administración & dosificación , Antiinfecciosos Locales/farmacocinética , Folículo Piloso/metabolismo , Miconazol/análogos & derivados , Piel/metabolismo , Administración Tópica , Adulto , Femenino , Estudios de Seguimiento , Humanos , Masculino , Miconazol/administración & dosificación , Miconazol/farmacocinética , Pruebas de Sensibilidad Microbiana/métodos , Absorción Cutánea , Adulto JovenRESUMEN
BACKGROUND: The skin represents a potent barrier to the environment, which can be enhanced by the topical application of skin care products, such as oil and oil-based formulations by moisturizing the skin. METHODS: The aim of this study was the investigation of the penetration behaviour of four vegetable oils and of paraffin oil into the stratum corneum by laser scanning microscopy. In addition, the occlusion capacity of these substances was assessed by transepidermal water loss (TEWL) measurements. Petrolatum served as a positive control for skin occlusion. The study was conducted in vivo and included six healthy volunteers. RESULTS: Paraffin oil, as well as the vegetable oils, penetrated only into the first upper layers of the stratum corneum. TEWL measurements indicated that the application of the vegetable oils (except jojoba oil) as well as paraffin oil, led to a similar occlusion of the skin surface. The most effective occlusion was found for petrolatum. CONCLUSION: For the investigated oils, a deeper penetration than into the first upper layers of the stratum corneum could be excluded. The decreased TEWL values indicate that the application of the oils leads to a semi-occlusion of the skin surface as it is intended by the use of oils to retain moisture in skin.
Asunto(s)
Parafina/farmacología , Parafina/farmacocinética , Aceites de Plantas/farmacología , Aceites de Plantas/farmacocinética , Absorción Cutánea/efectos de los fármacos , Absorción Cutánea/fisiología , Adulto , Humanos , Masculino , Persona de Mediana EdadRESUMEN
The pros and cons of the systemic and topical application of antioxidant substances are a subject of intense discussion among experts, with resulting confusion for consumers and producers. The objective of the present article is to clarify the various uncertainties relating to the use of antioxidant substances in dermatology. Whereas inappropriate application of antioxidant substances (concerning their concentration and composition) might induce harmful effects, the consumer will definitively benefit from physiological concentrations and compositions of antioxidants. The most suitable method is the consumption of natural antioxidants in the form of fruit and vegetables, for example. In addition, the skin, which also accumulates antioxidant substances, may profit from a sufficient antioxidative level, as damage induced by sun radiation in addition to skin aging is reduced.
Asunto(s)
Antioxidantes/administración & dosificación , Frutas , Estrés Oxidativo/efectos de los fármacos , Envejecimiento de la Piel/efectos de los fármacos , Piel/efectos de los fármacos , Verduras , Administración Tópica , Antioxidantes/farmacología , HumanosRESUMEN
In the case of topically applied substances, usually both lateral spreading and competitive penetration into the skin occur in parallel. In the present study, the pathways of lateral spreading were studied quantitatively and visually. The local distribution and lateral spreading of the UV filter substance butyl methoxydibenzoylmethane applied in an o/w emulsion was studied on the forearm and the back. The tape stripping procedure was used to determine the recovery rates inside and outside the area of application. The skin characteristics of transepidermal water loss, pH value, hydration of the stratum corneum and sebum rate were determined at both anatomic sites. Photography and laser scanning microscopy were used to visually investigate the lateral spreading of topically applied dyes. On the back, a preferred direction of lateral spreading parallel to the body axis was observed. This result was caused by differences in the network of furrows. The furrows functioned as a pathway for lateral spreading, whereas the follicles formed a reservoir for the topically applied substance.
Asunto(s)
Absorción Cutánea , Protectores Solares/farmacocinética , Administración Cutánea , Adulto , Alcanos/administración & dosificación , Alcanos/análisis , Dorso , Chalconas/administración & dosificación , Chalconas/análisis , Emulsiones , Femenino , Antebrazo , Humanos , Concentración de Iones de Hidrógeno , Masculino , Propiofenonas , Piel/metabolismo , Análisis Espectral , Protectores Solares/administración & dosificación , Cinta Quirúrgica , Pérdida Insensible de Agua , Adulto JovenRESUMEN
Environmental factors like air pollutants, radiation of the sun and stress factors such as illness, smoking, or alcohol abuse produce free radicals in the human tissue as well as in the skin. Free radicals serve as the main cause for premature skin aging. Additionally, they also contribute towards immunosuppression and the formation of skin diseases including cancer. The human organism has developed a protection system against the destructive action of free radicals by means of the antioxidant network. In the present study, the interaction of free radicals and carotenoid antioxidants in the human skin under in vivo conditions was investigated and summarized. The measurement of carotenoids in human skin was performed in vivo using resonance Raman spectroscopy.
Asunto(s)
Carotenoides/metabolismo , Radicales Libres/metabolismo , Envejecimiento de la Piel/efectos de los fármacos , Piel/metabolismo , Adulto , Antioxidantes , Humanos , Rayos Infrarrojos , Persona de Mediana Edad , Piel/efectos de la radiación , Espectrometría Raman , Rayos Ultravioleta , Adulto JovenRESUMEN
The hair follicles could be a reservoir for topically applied substances. They are not only surrounded by a close network of blood capillaries, which makes them interesting for drug delivery, but they are also the host of dendritic cells, which are important for immunomodulation. Previously, pollen allergens were shown to penetrate into the hair follicles. The application of barrier-enhancing formulations might represent an effective strategy to prevent pollen protein penetration into the hair follicle. In the present study, porcine skin areas were pretreated with 4 barrier-enhancing emulsions. One skin area served as control and remained without pretreatment. Afterwards, fluorescein-isothiocyanate-labeled grass pollen proteins were applied to the porcine skin samples, and their penetration was investigated via fluorescent laser scanning microscopy. It was shown that the barrier-enhancing formulations were able to significantly reduce the penetration of exogenous proteins into the hair follicles, the extent of such reduction depending on the formulation.
Asunto(s)
Alérgenos/farmacocinética , Folículo Piloso/metabolismo , Polen/inmunología , Absorción Cutánea/efectos de los fármacos , Alérgenos/inmunología , Animales , Emulsiones , Fluoresceína-5-Isotiocianato/química , Colorantes Fluorescentes/química , Lípidos/administración & dosificación , Lípidos/farmacología , Poaceae/inmunología , PorcinosRESUMEN
The application of particles in dermatology and cosmetology represents an emerging field and is closely connected with the question of risk assessment as the potential for, and consequences of, penetration of such particles into the living tissue has not been determined conclusively. In the medical sector, extensive research activities are in progress to develop particles, which can be used as efficient carriers for drug delivery through the skin barrier. In contrast, in cosmetic products, particles are mostly required to remain on the skin surface to fulfill their beneficial effect. Whereas the intercellular penetration of particles seems to be unlikely, the hair follicle has been shown to be a relevant penetration pathway for particles as well as an important long-term reservoir. It has been demonstrated that the penetration depth of the particles can be influenced by their size resulting in the possibility of a differentiated targeting of specific follicular structures. In the present review, the follicular penetration mechanisms and storage properties of particles are discussed.
Asunto(s)
Cosméticos/farmacocinética , Nanopartículas/administración & dosificación , Piel/metabolismo , Animales , Transporte Biológico , Cosméticos/administración & dosificación , Cosméticos/efectos adversos , Sistemas de Liberación de Medicamentos , Folículo Piloso/efectos de los fármacos , Folículo Piloso/metabolismo , Humanos , Nanopartículas/efectos adversos , Tamaño de la Partícula , Permeabilidad , Piel/efectos de los fármacos , Absorción CutáneaRESUMEN
Stimulation of the penetration of topically applied substances into the skin is a topic of intensive dermatological and pharmacological research. Next to intercellular penetration, i.e. a penetration inside the lipid layers around the corneocytes, follicular penetration also represents an efficient penetration pathway. The hair follicles act as a long-term reservoir for topically applied substances. They are surrounded by or contain several important target structures, such as blood capillaries, stem cells and dendritic cells. Therefore, the hair follicles have to be well protected from hazardous substances coming into contact with the skin. The traditional method of decontamination of the skin involves an intensive washing procedure. However, this process represents a massage, which pushes the hazardous substances even deeper into the hair follicles. In the present study, the application of absorbing materials for decontamination of the skin was investigated after the application of a model substance utilizing the tape-stripping procedure. It was found that absorbing materials are better suited than the washing process for decontamination of the skin.
Asunto(s)
Cinamatos/metabolismo , Descontaminación/métodos , Folículo Piloso/metabolismo , Poliuretanos/administración & dosificación , Absorción Cutánea , Protectores Solares/metabolismo , Irrigación Terapéutica , Administración Cutánea , Adulto , Anciano , Cinamatos/administración & dosificación , Antebrazo , Humanos , Persona de Mediana Edad , Nanofibras , Protectores Solares/administración & dosificación , Adulto JovenRESUMEN
Tape stripping is a standard measuring method for the investigation of the dermatopharmacokinetics of topically applied substances using adhesive films. These tape strips are successively applied and removed from the skin after application and penetration of topically applied substances. Thus, layers of corneocytes and some amount of topical applied substances are removed. The amount of substances and the amount of stratum corneum removed with a single tape strip has to be determined for the calculation of the penetration profile. The topically applied substances removed from the skin can be determined by classical analytical methods like high-pressure liquid chromatography, mass spectroscopy, and spectroscopic measurements. The amount of corneocytes on the tape strips can be easily detected by their pseudoabsorption. In the present paper, an easy and cheap corneocyte density analyzer is presented that is based on a slide projector. Comparing the results of the measurements obtained by the corneocyte density analyzer and by uv-visible spectrometry, identical results were obtained.
Asunto(s)
Recuento de Células/instrumentación , Clobetasol/administración & dosificación , Clobetasol/farmacocinética , Queratinocitos/citología , Queratinocitos/metabolismo , Absorción Cutánea/fisiología , Cinta Quirúrgica , Administración Tópica , Adulto , Recuento de Células/métodos , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Óptica y Fotónica/instrumentación , Fotometría/instrumentación , Fotometría/métodosRESUMEN
Reactive free radicals can be produced in the skin by the action of environmental factors, such as sun radiation and toxins. These radicals can damage the DNA, proteins and lipids of the living cells. The consequences can be skin aging, immune suppression and even skin cancer. Humans have developed a protective mechanism against the action of free radicals in the form of antioxidant substances. Several of these antioxidants cannot be produced by humans and have to be acquired via food, such as carotenoids. Optical, non-invasive methods, like resonance Raman spectroscopy, allow a qualitative and quantitative online detection of the kinetics of antioxidants such as carotenoids in the skin. By employing this method it has been shown that the uptake of carotenoids in food can lead to an accumulation in the skin. On the other hand, stress, illness and UV-radiation can reduce the concentration of antioxidant substances in the skin. A high concentration of antioxidant substances is protective and associated with a reduction in skin wrinkling.
Asunto(s)
Antioxidantes/administración & dosificación , Fenómenos Fisiológicos de la Piel , Antioxidantes/farmacocinética , Bebidas , Disponibilidad Biológica , Carotenoides/administración & dosificación , Carotenoides/farmacocinética , Dermatitis por Contacto/fisiopatología , Dermatitis por Contacto/prevención & control , Alimentos , Humanos , Licopeno , Valor Nutritivo , Procesamiento de Señales Asistido por Computador , Envejecimiento de la Piel/efectos de los fármacos , Envejecimiento de la Piel/efectos de la radiación , Espectrometría Raman/instrumentación , Quemadura Solar/fisiopatología , Quemadura Solar/prevención & control , Rayos Ultravioleta/efectos adversos , Verduras , beta Caroteno/administración & dosificación , beta Caroteno/farmacocinéticaRESUMEN
The objective and quantitative application of tape stripping in pharmaceutics and dermatopharmacokinetics requires the determination of the exact position of each removed tape strip inside the stratum corneum (SC) and/or the determination of the relative SC thickness. In this study, transepidermal water loss (TEWL) and the optical spectroscopic data of the corneocytes were measured simultaneously during the complete removal of the SC by tape stripping. The spectroscopic data quantitatively reflect the amount of corneocytes removed by the individual tape strips, whereas TEWL and 1/TEWL are not sensitive enough to measure the relatively small changes in the SC thickness realized by the removal of the individual strips. The relative SC thickness can be determined directly by the spectroscopic data, while the 1/TEWL values require a second independent method. The results demonstrate the importance of tape stripping characterizing the behaviour of topically applied substances.